Sample records for active site gorge

  1. Flexibility of active-site gorge aromatic residues and non-gorge aromatic residues in acetylcholinesterase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghattyvenkatakrishna, Pavan K; Uberbacher, Edward C

    2013-01-01

    The presence of an unusually large number of aromatic residues in the active site gorge of acetylcholinesterase has been a topic of great interest. Flexibility of these residues has been suspected to be a key player in controlling ligand traversal in the gorge. This raises the question of whether the over representation of aromatic residues in the gorge implies higher than normal flexibility of those residues. The current study suggests that it does not. Large changes in the hydrophobic cross sectional area due to dihedral oscillations are probably the reason behind their presence in the gorge.

  2. Analysis of the activation of acetylcholinesterase by carbon nanoparticles using a monolithic immobilized enzyme microreactor: role of the water molecules in the active site gorge.

    PubMed

    Ibrahim, Firas; Andre, Claire; Iutzeler, Anne; Guillaume, Yves Claude

    2013-10-01

    A biochromatographic system was used to study the direct effect of carbon nanoparticles (CNPs) on the acetylcholinesterase (AChE) activity. The AChE enzyme was covalently immobilized on a monolithic CIM-disk via its NH2 residues. Our results showed an increase in the AChE activity in presence of CNPs. The catalytic constant (k(cat)) was increased while the Michaelis constant (K(m)) was slightly decreased. This indicated an increase in the enzyme efficiency with increase of the substrate affinity to the active site. The thermodynamic data of the activation mechanism of the enzyme, i.e. ΔH* and ΔS*, showed no change in the substrate interaction mechanism with the anionic binding site. The increase of the enthalpy (ΔH*) and the entropy (ΔS*) with decrease in the free energy of activation (Ea) was related to structural conformation change in the active site gorge. This affected the stability of water molecules in the active site gorge and facilitated water displacement by substrate for entering to the active site of the enzyme.

  3. Active tectonics at the Lower Yarmouk Gorge?

    NASA Astrophysics Data System (ADS)

    Inbar, Nimrod; Raggad, Marwan; Siebert, Christian; Möller, Peter; Rödiger, Tino; Rosenthal, Eliyaho; Guttman, Josseph; Magri, Fabien

    2017-04-01

    The Lower Yarmouk Gorge (LYG) extends on the eastern margin of the Jordan Rift Valley (JRV) adjacent to the Kinneret basin which makes part in the chain of pull-apart basins along the Dead Sea Transform (DST). The LYG is bounded to the south by the Ajloun Plateau (northern Jordan) and to the north by the Golan Heights (Israel). It acts as the outflow of the Yarmouk drainage basin into the Jordan River a few kilometers south of Lake Tiberias. Although topographically the Golan Heights and the Ajloun seem to be different provinces separated by the LYG, the northern Ajloun and southern Golan form the southern flank of a major synclinal structure. Morphologically, the LYG resembles the Sheikh-Ali strike-slip fault and other NE-SW striking faults related to the DST. However, the existence of faulting along that line is debatable. As the gorge serves also as state boundary, no seismic lines cross the LYG. Quaternary landslides, mostly on the southern flank of the LYG cover possible surface evidence of faulting although Quaternary basalts located at the gorge path may indicate possible vertical pathways. Moreover, hydrological studies (Siebert et al., 2014, Goretzki et al., 2016) show that permeability anisotropy along the LYG line allows heated groundwater to emerge along the gorge with temperatures rising up to 60°C. The presented study uses well data from northern Jordan and southern Golan Heights as well as seismic data from the southern Golan Heights to bridge over the information gap. Based on the available information we present our hypothesis on the geology of the LYG aiming to contribute to the discussion regarding active tectonics at the Lower Yarmouk Gorge. Our preliminary results show that a major fault along the gorge path is dubious however, perpendicular faulting is more likely to occur. We expect the results of current research to contribute to the understanding of the local complex geohydrological system. Furthermore, results are expected to deepen our

  4. Three Gorges Dam, China

    NASA Image and Video Library

    2002-07-25

    This ASTER image shows a 60 km stretch of the Yangtze River in China, including the Xiling Gorge, the eastern of the three gorges. In the left part of the image is the construction site of the Three Gorges Dam, the world's largest. This image was acquired on July 20, 2000 by the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) on NASA's Terra satellite. With its 14 spectral bands from the visible to the thermal infrared wavelength region, and its high spatial resolution of 15 to 90 meters (about 50 to 300 feet), ASTER will image Earth for the next 6 years to map and monitor the changing surface of our planet. http://photojournal.jpl.nasa.gov/catalog/PIA03852

  5. The role of the peripheral anionic site and cation-pi interactions in the ligand penetration of the human AChE gorge.

    PubMed

    Branduardi, Davide; Gervasio, Francesco Luigi; Cavalli, Andrea; Recanatini, Maurizio; Parrinello, Michele

    2005-06-29

    We study the ligand (tetramethylammonium) recognition by the peripheral anionic site and its penetration of the human AChE gorge by using atomistic molecular dynamics simulations and our recently developed metadynamics method. The role of both the peripheral anionic site and the formation of cation-pi interactions in the ligand entrance are clearly shown. In particular, a simulation with the W286A mutant shows the fundamental role of this residue in anchoring the ligand at the peripheral anionic site of the enzyme and in positioning it prior to the gorge entrance. Once the ligand is properly oriented, the formation of specific and synchronized cation-pi interactions with W86, F295, and Y341 enables the gorge penetration. Eventually, the ligand is stabilized in a free energy basin by means of cation-pi interactions with W86.

  6. Exploring quality standards for New River Gorge climbing sites: establishing a baseline for the future

    Treesearch

    Roy Ramthun; Andy Blake

    2003-01-01

    The New River Gorge National River has become a major destination for sport climbers in the eastern U.S. A new climbing management plan is being developed for the site. This study examined the satisfaction levels of climbers with some of the easily managed facilities at climbing sites and looked at attributes of the social setting preferred by climbers. Climbers at the...

  7. Dynamic Mechanism of a Fluorinated Oxime Reactivator Unbinding from AChE Gorge in Polarizable Water.

    PubMed

    Pathak, Arup K; Bandyopadhyay, Tusar

    2018-04-12

    A well-tempered metadynamics simulation is performed to study the unbinding process of a fluorinated oxime (FHI-6) drug from the active-site gorge of acetylcholinesterase enzyme in a polarizable water medium. Cation-π interactions and water bridge and hydrogen bridge formations between the protein and the drug molecule are found to strongly influence the unbinding process, forming basins and barriers along the gorge pathway. Distinct unbinding pathways are found when FHI-6 was compared with its recently reported nonfluorinated analogue, HI-6. For example, because of permanent positive charges on both the pyridinium rings of HI-6, it exhibits the minimum in the potential of mean force of the unbinding process in the gorge mouth (where the peripheral anion site, PAS, of the enzyme is located), which is largely caused by cation-π interactions. However, the same interaction, both in the catalytic active-site (CAS) and PAS regions, is found to be greatly enhanced in its lipophilic fluorinated analogue, FHI-6, causing a deep potential energy minimum in the bound state. This may render FHI-6 to be held more firmly in the CAS region of the gorge, as is also evidenced from the microkinetics of unbinding transitions, measured through a combination of metadynamics and hyperdynamics simulations.

  8. 2. GORGE HIGH DAM. UNUSUALLY HIGH WATER IN GORGE LAKE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. GORGE HIGH DAM. UNUSUALLY HIGH WATER IN GORGE LAKE DUE TO THE COMBINATION OF UNIT 24 BEING DOWN FOR REWINDING AND TWO UNITS COMING ON LINE UNEXPECTEDLY AT ROSS POWERHOUSE LED TO WATER FLOWING OVER THE SPILLGATES. EACH GATE IF 47 FEET WIDE AND 50 FEET HIGH, 1989. - Skagit Power Development, Gorge High Dam, On Skagit River, 2.9 miles upstream from Newhalem, Newhalem, Whatcom County, WA

  9. How does huperzine A enter and leave the binding gorge of acetylcholinesterase? Steered molecular dynamics simulations.

    PubMed

    Xu, Yechun; Shen, Jianhua; Luo, Xiaomin; Silman, Israel; Sussman, Joel L; Chen, Kaixian; Jiang, Hualiang

    2003-09-17

    The entering and leaving processes of Huperzine A (HupA) binding with the long active-site gorge of Torpedo californica acetylcholinesterase (TcAChE) have been investigated by using steered molecular dynamics simulations. The analysis of the force required along the pathway shows that it is easier for HupA to bind to the active site of AChE than to disassociate from it, which for the first time interprets at the atomic level the previous experimental result that unbinding process of HupA is much slower than its binding process to AChE. The direct hydrogen bonds, water bridges, and hydrophobic interactions were analyzed during two steered molecular dynamics (SMD) simulations. Break of the direct hydrogen bond needs a great pulling force. The steric hindrance of bottleneck might be the most important factor to produce the maximal rupture force for HupA to leave the binding site but it has a little effect on the binding process of HupA with AChE. Residue Asp72 forms a lot of water bridges with HupA leaving and entering the AChE binding gorge, acting as a clamp to take out HupA from or put HupA into the active site. The flip of the peptide bond between Gly117 and Gly118 has been detected during both the conventional MD and SMD simulations. The simulation results indicate that this flip phenomenon could be an intrinsic property of AChE and the Gly117-Gly118 peptide bond in both HupA bound and unbound AChE structures tends to adopt the native enzyme structure. At last, in a vacuum the rupture force is increased up to 1500 pN while in water solution the greatest rupture force is about 800 pN, which means water molecules in the binding gorge act as lubricant to facilitate HupA entering or leaving the binding gorge.

  10. Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate.

    PubMed

    Johnson, Joseph L; Cusack, Bernadette; Davies, Matthew P; Fauq, Abdul; Rosenberry, Terrone L

    2003-05-13

    Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3-(acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild-type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate

  11. [Effects of altitudes on soil microbial biomass and enzyme activity in alpine-gorge regions.

    PubMed

    Cao, Rui; Wu, Fu Zhong; Yang, Wan Qin; Xu, Zhen Feng; Tani, Bo; Wang, Bin; Li, Jun; Chang, Chen Hui

    2016-04-22

    In order to understand the variations of soil microbial biomass and soil enzyme activities with the change of altitude, a field incubation was conducted in dry valley, ecotone between dry valley and mountain forest, subalpine coniferous forest, alpine forest and alpine meadow from 1563 m to 3994 m of altitude in the alpine-gorge region of western Sichuan. The microbial biomass carbon and nitrogen, and the activities of invertase, urease and acid phosphorus were measured in both soil organic layer and mineral soil layer. Both the soil microbial biomass and soil enzyme activities showed the similar tendency in soil organic layer. They increased from 2158 m to 3028 m, then decreased to the lowest value at 3593 m, and thereafter increased until 3994 m in the alpine-gorge region. In contrast, the soil microbial biomass and soil enzyme activities in mineral soil layer showed the trends as, the subalpine forest at 3028 m > alpine meadow at 3994 m > montane forest ecotone at 2158 m > alpine forest at 3593 m > dry valley at 1563 m. Regardless of altitudes, soil microbial biomass and soil enzyme activities were significantly higher in soil organic layer than in mineral soil layer. The soil microbial biomass was significantly positively correlated with the activities of the measured soil enzymes. Moreover, both the soil microbial biomass and soil enzyme activities were significantly positively correlated with soil water content, organic carbon, and total nitrogen. The activity of soil invertase was significantly positively correlated with soil phosphorus content, and the soil acid phosphatase was so with soil phosphorus content and soil temperature. In brief, changes in vegetation and other environmental factors resulting from altitude change might have strong effects on soil biochemical properties in the alpine-gorge region.

  12. Three Gorges Dam, China

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This ASTER image shows a 60 km stretch of the Yangtze River in China, including the Xiling Gorge, the eastern of the three gorges. In the left part of the image is the construction site of the Three Gorges Dam, the world's largest.

    This image was acquired on July 20, 2000 by the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) on NASA's Terra satellite. With its 14 spectral bands from the visible to the thermal infrared wavelength region, and its high spatial resolution of 15 to 90 meters (about 50 to 300 feet), ASTER will image Earth for the next 6 years to map and monitor the changing surface of our planet.

    ASTER is one of five Earth-observing instruments launched December 18, 1999, on NASA's Terra satellite. The instrument was built by Japan's Ministry of Economy, Trade and Industry. A joint U.S./Japan science team is responsible for validation and calibration of the instrument and the data products. Dr. Anne Kahle at NASA's Jet Propulsion Laboratory, Pasadena, California, is the U.S. Science team leader; Bjorn Eng of JPL is the project manager. The Terra mission is part of NASA's Earth Science Enterprise, a long-term research and technology program designed to examine Earth's land, oceans, atmosphere, ice and life as a total integrated system.

    The broad spectral coverage and high spectral resolution of ASTER will provide scientists in numerous disciplines with critical information for surface mapping, and monitoring dynamic conditions and temporal change. Example applications are: monitoring glacial advances and retreats; monitoring potentially active volcanoes; identifying crop stress; determining cloud morphology and physical properties; wetlands evaluation; thermal pollution monitoring; coral reef degradation; surface temperature mapping of soils and geology; and measuring surface heat balance.

    Size: 60 x 24 km (36 x 15 miles) Location: 30.6 deg. North lat., 111.2 deg. East long. Orientation: North at top Image Data: ASTER

  13. Dating the Naisiusiu Beds, Olduvai Gorge, by electron spin resonance

    NASA Astrophysics Data System (ADS)

    Skinner, A. R.; Hay, R. L.; Masao, F.; Blackwell, B. A. B.

    2003-05-01

    The lower beds at Olduvai Gorge are well known for containing early hominid fossils and Oldowan stone tools, and their ages have been established by 40Ar/ 39Ar dating and paleomagnetic stratigraphy. Ages are generally less certain for the upper deposits at Olduvai Gorge because of the scarcity of datable tuffs. The youngest archaeologically significant site at Olduvai is microlithic LSA, which lies in the type section of the Naisiusiu Beds. The age for the site is controversial, with 14C dates of 17,000-17,550 (Hay, R.L., 1976 Geology of Olduvai Gorge, University of California Press, Berkeley) and >42,000 BP (Manega, P.C., 1993. Geochronology, geochemistry, and isotopic study of the Plio-Pleistocene Hominid sites and the Ngorongoro Volcanic Highland in Northern Tanzania. Unpublished Ph.D. Thesis, University of Colorado, Boulder, CO). The tuff bed in the zone with artifacts does not contain materials datable by 40Ar/ 39Ar, and some other dating method was needed. In the summer of 2001, five equid teeth were collected from the type Naisiusiu site. Another tooth had previously been collected. ESR ages have been determined for three teeth from the archaeological level and their ages cluster around 62±5 ka, assuming linear uranium uptake. Another tooth from a level without artifacts and believed to be significantly younger dated to 39±5 ka, again assuming LU. These dates are considerably older than previous estimates and suggest that the East African MSA/LSA transition occurred very early.

  14. Erosion of the Yarlung-Tsangpo Gorge sustained by episodic megaflooding

    NASA Astrophysics Data System (ADS)

    Lang, K. A.; Huntington, K. W.

    2012-12-01

    As the Yarlung-Tsangpo river descends through the easternmost Himalaya to join the Brahmaputra river in India, it carves a ~2 km-deep knickzone: the Yarlung-Tsangpo Gorge. Within the gorge, fluvial incision has kept pace with an active crustal scale antiform, exhuming mid-crustal material over Pliocene time at rates up to 7-10 km/Myr. This close spatial correspondence of erosion and exhumation has led previous researchers to hypothesize a self-perpetuating relationship between focused erosion and rock uplift in this ~1200 km^2 region. While high discharge in the river today produces bed stresses that may have the erosional capability to match rock uplift rates, this hypothesis also requires erosion to be localized in the gorge region over timescales spanning multiple glacial periods during which the upstream drainage was episodically dammed by glaciers and discharge was variable. We interpret downstream flood deposits corresponding to this episodic damming as direct evidence of megaflood action as the principal agent of gorge erosion, spanning multiple glacial periods and potentially matching the million-year exhumation rates independently measured in the gorge. Previous researchers documented evidence of glacial damming throughout the fluvial network upstream of the gorge, where moraine and landslide dams are abundant, often beneath dissected lake deposits. We report flood deposits downstream of the gorge, remaining along valley walls at multiple elevations up to 150 m off the modern channel. Using LA-ICPMS U-Pb dating of detrital zircons from these flood sediments, we demonstrate that these high-magnitude flood events disproportionately excavated material from the Yarlung-Tsangpo Gorge, and propose an alternative erosional mechanism to match rock uplift in the gorge over a 10^6 year timescale. We analyzed five flood deposits spanning a range of elevations above the modern channel and four deposits from a documented lower-magnitude flood in 2000. We constrain

  15. Nutritional and behavioral effects of gorge and fast feeding in captive lions.

    PubMed

    Altman, Joanne D; Gross, Kathy L; Lowry, Stephen R

    2005-01-01

    Nonhuman animals in captivity manifest behaviors and physiological conditions that are not common in the wild. Lions in captivity face problems of obesity, inactivity, and stereotypy. To mediate common problems of captive lions, this study implemented a gorge and fast feeding schedule that better models naturalistic patterns: African lions (Panthera leo) gradually adapted from a conventional feeding program to a random gorge and fast feeding schedule. Digestibility increased significantly and food intake and metabolizable energy intake correspondingly decreased. Lions also showed an increase in appetitive active behaviors, no increase in agonistic behavior, and paced half as frequently on fast days as on feeding days. Thus, switching captive lions to a gorge and fast feeding schedule resulted in improved nutritional status and increased activity.

  16. Influence of filling-drawdown cycles of the Three Gorges reservoir on deformation and failure behaviors of anaclinal rock slopes in the Wu Gorge

    NASA Astrophysics Data System (ADS)

    Huang, Da; Gu, Dong Ming

    2017-10-01

    The upper Wu Gorge on the Yangtze River has been the site of tens of reservoir-induced landslides since the filling of the Three Gorges reservoir in 2003. These landslides have been occurring in heavily fractured carbonate rock materials along the rim of the reservoir in the Wu Gorge. A detailed investigation was carried out to examine the influence of reservoir operations (filling and drawdown) on slope stabilities in the upper Wu Gorge. Field investigations reveal many collapses of various types occurred at the toe of the anaclinal rock slopes, owing to the long-term intensive river erosion caused by periodic fluctuation of the reservoir level. Analysis of data from deformation monitoring suggests that the temporal movement of the slopes shows seasonal fluctuations that correlate with reservoir levels and drawdown conditions, with induced slope acceleration peaking when reservoir levels are lowest. This may illustrate that the main mechanism is the reservoir drawdown, which induces an episodic seepage force in the highly permeable materials at the slope toes, and thus leads to the episodic rockslides. The coupled hydraulic-mechanical (HM) modeling of the G2 landslide, which occurred in 2008, shows that collapse initiated at the submerged slope toe, which then caused the upper slope to collapse in a rock topple-rock slide pattern. The results imply that preventing water erosion at the slope toe might be an effective way for landslide prevention in the study area.

  17. Geomorphological evolution of a fluvial channel after primary lahar deposition: Huiloac Gorge, Popocatépetl volcano (Mexico)

    NASA Astrophysics Data System (ADS)

    Tanarro, L. M.; Andrés, N.; Zamorano, J. J.; Palacios, D.; Renschler, C. S.

    2010-10-01

    Popocatépetl volcano (19°02' N, 98°62' W, 5424 m) began its most recent period of volcanic activity in December 1994. The interaction of volcanic and glacier activity triggered the formation of lahars through the Huiloac Gorge, located on the northern flank of the volcano, causing significant morphological changes in the channel. The most powerful lahars occurred in April 1995, July 1997 and January 2001, and were followed by secondary lahars that formed during the post-eruptive period. This study interprets the geomorphological evolution of the Huiloac Gorge after the January 2001 lahar. Variations in channel morphology at a 520 m-long research site located mid-way down the gorge were recorded over a 4 year period from February 2002 to March 2005, and depicted in five geomorphological maps (scale 1:200) for 14 February and 15 October 2002, 27 September 2003, 9 February 2004, and 16 March 2006. A GIS was used to calculate the surface area for the landforms identified for each map and detected changes and erosion-deposition processes of the landforms using the overlay function for different dates. Findings reveal that secondary lahars and others types of flows, like sediment-laden or muddy streamflows caused by precipitation, rapidly modified the gorge channel following the January 2001 non-eruptive lahar, a period associated with volcanic inactivity and the disappearance of the glacier once located at the headwall of the gorge. Field observations also confirmed that secondary flows altered the dynamics and geomorphological development of the channel. These flows incised and destroyed the formations generated by the primary lahars (1997 and 2001), causing a widening of the channel that continues today. After February 2004, a rain-triggered lahar and other flows infilled the channel with materials transported by these flows. The deposits on the lateral edges of the channel form terraces. A recent lull in lahar activity contrasts with the increasing instability of

  18. The contexts and early Acheulean archaeology of the EF-HR paleo-landscape (Olduvai Gorge, Tanzania).

    PubMed

    de la Torre, Ignacio; Albert, Rosa M; Macphail, Richard; McHenry, Lindsay J; Pante, Michael C; Rodríguez-Cintas, Ágata; Stanistreet, Ian G; Stollhofen, Harald

    2017-08-03

    Renewed fieldwork at the early Acheulean site of EF-HR (Olduvai Gorge, Tanzania) has included detailed stratigraphic studies of the sequence, extended excavations in the main site, and has placed eleven additional trenches within an area of nearly 1 km 2 , to sample the same stratigraphic interval as in the main trench across the broader paleo-landscape. Our new stratigraphic work suggests that EF-HR is positioned higher in the Bed II sequence than previously proposed, which has implications for the age of the site and its stratigraphic correlation to other Olduvai Middle Bed II sites. Geological research shows that the main EF-HR site was situated at the deepest part of an incised valley formed through river erosion. Archaeological excavations at the main site and nearby trenches have unearthed a large new assemblage, with more than 3000 fossils and artefacts, including a hundred handaxes in stratigraphic position. In addition, our test-trenching approach has detected conspicuous differences in the density of artefacts across the landscape, with a large cluster of archaeological material in and around the main trench, and less intense human activity at the same level in the more distant satellite trenches. All of these aspects are discussed in this paper in the light of site formation processes, behavioral contexts, and their implications for our understanding of the early Acheulean at Olduvai Gorge. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Dietary traits of the ungulates from the HWK EE site at Olduvai Gorge (Tanzania): Diachronic changes and seasonality.

    PubMed

    Rivals, Florent; Uno, Kevin T; Bibi, Faysal; Pante, Michael C; Njau, Jackson; de la Torre, Ignacio

    2017-09-01

    The Oldowan site HWK EE (Olduvai Gorge, Tanzania) has yielded a large fossil and stone tool assemblage at the transition from Lower to Middle Bed II, ∼1.7 Ma. Integrated tooth wear and stable isotope analyses were performed on the three most abundant ungulate taxa from HWK EE, namely Alcelaphini, cf. Antidorcas recki (Antilopini) and Equus oldowayensis (Equini), to infer dietary traits in each taxon. Some paleodietary changes were observed for cf. A. recki and E. oldowayensis based on tooth wear at the transition from the Lemuta to the Lower Augitic Sandstone (LAS) interval within the HWK EE sequence. Stable carbon and oxygen isotope data show no significant changes in bulk diet or hydroclimate between the Lemuta and LAS intervals. The combined tooth wear and stable isotope data suggest similar paleoecological conditions across the two HWK EE intervals, but that differences in vegetation consumed among ungulates may have resulted in changes in dietary niches. Integrating tooth wear and stable isotope analyses permits the characterization of ungulate diets and habitats at HWK EE where C 4 dominated and minor mixed C 3 and C 4 habitats were present. Our results provide a better understanding of the paleoenvironmental conditions of the Lemuta and LAS intervals. The LAS assemblage was mostly accumulated during relatively dry periods at Olduvai Gorge when grasses were not as readily available and grazing animals may have been more nutritionally-stressed than during the formation of the Lemuta assemblage. This helps to contextualize variations in hominin and carnivore feeding behavior observed from the faunal assemblages produced during the two main occupations of the site. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Wetland plant waxes from Olduvai Gorge, Tanzania

    NASA Astrophysics Data System (ADS)

    Tamalavage, A.; Magill, C. R.; Barboni, D.; Ashley, G. M.; Freeman, K. H.

    2013-12-01

    Olduvai Gorge, northern Tanzania, exposes a Plio-Pleistocene sedimentary record that includes lake and lake-margin sediments and fossil remains of ancient plants and early humans. There are rich paleontological and cultural records at Olduvai Gorge that include thousands of vertebrate fossils and stone tools. Previous studies of plant biomarkers in lake sediments from Olduvai Gorge reveal repeated, abrupt changes in landscape dominance by woodland or grassland vegetation during the early Pleistocene, about 1.8 million years ago. However, the reconstruction of wetland vegetation in the past is limited by a dearth of published lipid signatures for modern wetland species. Here, we present lipid and isotopic data for leaf tissues from eight modern plants (i.e., sedge and Typha species) living in wetlands near Olduvai Gorge. Trends in values for molecular and leaf δ13C and average chain length (ACL) of n-alkanes in plant tissues are similar to values for underlying soils. Compound-specific δ13C values for n-alkanes C25 to C33 range between -36.4 to -23.1‰ for C3 plants and -22.3 to -19.5‰ for C4 plants. Fractionation factors between leaf and lipids, ɛ29 and ɛ33, fall within the range reported in the literature, but they differ more widely within a single plant. For C3 plants, the average difference between ɛ29 and ɛ33 is 6.5 ‰, and the difference between ɛ29 and ɛ33 for C4 plants is less than 2‰. Both plant types show a parabolic relationship between chain length and δ13C values, in which C29 typically has the most depleted value, and typically shift by 3-5‰ between alkane homologs. This pattern has not been previously reported, and could be unique for sedge lipids. If so, these data help constrain the application of plant wax biomarkers from sedges for paleo-vegetation reconstruction in paleoclimate studies and at archaeological sites.

  1. Long Valley Caldera Lake and reincision of Owens River Gorge

    USGS Publications Warehouse

    Hildreth, Wes; Fierstein, Judy

    2016-12-16

    Owens River Gorge, today rimmed exclusively in 767-ka Bishop Tuff, was first cut during the Neogene through a ridge of Triassic granodiorite to a depth as great as its present-day floor and was then filled to its rim by a small basaltic shield at 3.3 Ma. The gorge-filling basalt, 200 m thick, blocked a 5-km-long reach of the upper gorge, diverting the Owens River southward around the shield into Rock Creek where another 200-m-deep gorge was cut through the same basement ridge. Much later, during Marine Isotope Stage (MIS) 22 (~900–866 ka), a piedmont glacier buried the diversion and deposited a thick sheet of Sherwin Till atop the basalt on both sides of the original gorge, showing that the basalt-filled reach had not, by then, been reexcavated. At 767 ka, eruption of the Bishop Tuff blanketed the landscape with welded ignimbrite, deeply covering the till, basalt, and granodiorite and completely filling all additional reaches of both Rock Creek canyon and Owens River Gorge. The ignimbrite rests directly on the basalt and till along the walls of Owens Gorge, but nowhere was it inset against either, showing that the basalt-blocked reach had still not been reexcavated. Subsidence of Long Valley Caldera at 767 ka produced a steep-walled depression at least 700 m deeper than the precaldera floor of Owens Gorge, which was beheaded at the caldera’s southeast rim. Caldera collapse reoriented proximal drainages that had formerly joined east-flowing Owens River, abruptly reversing flow westward into the caldera. It took 600,000 years of sedimentation in the 26-km-long, usually shallow, caldera lake to fill the deep basin and raise lake level to its threshold for overflow. Not until then did reestablishment of Owens River Gorge begin, by incision of the gorge-filling ignimbrite.

  2. MIDDLE GORGE POWER PLANT, OWENS RIVER STREAM FLOWING OVER TAIL ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    MIDDLE GORGE POWER PLANT, OWENS RIVER STREAM FLOWING OVER TAIL RACE OF POWER PLANT AND PENSTOCK HEADGATE TO LOWER GORGE CONTROL PLANT. A MINIMAL FLOW OF RIVER WATER IS REQUIRED TO MAINTAIN FISH LIFE - Los Angeles Aqueduct, Middle Gorge Power Plant, Los Angeles, Los Angeles County, CA

  3. 1. GORGE HIGH DAM. THIS THIN ARCH DAM WITH A ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. GORGE HIGH DAM. THIS THIN ARCH DAM WITH A GRAVITY SECTION IS THE THIRD DAM BUILT BY SEATTLE CITY LIGHT TO PROVIDE WATER FOR GORGE POWERHOUSE AND WAS COMPLETED IN 1961, 1989. - Skagit Power Development, Gorge High Dam, On Skagit River, 2.9 miles upstream from Newhalem, Newhalem, Whatcom County, WA

  4. The Three Gorges Project: How sustainable?

    NASA Astrophysics Data System (ADS)

    Kepa Brian Morgan, Te Kipa; Sardelic, Daniel N.; Waretini, Amaria F.

    2012-08-01

    SummaryIn 1984 the Government of China approved the decision to construct the Three Gorges Dam Project, the largest project since the Great Wall. The project had many barriers to overcome, and the decision was made at a time when sustainability was a relatively unknown concept. The decision to construct the Three Gorges Project remains contentious today, especially since Deputy Director of the Three Gorges Project Construction Committee, Wang Xiaofeng, stated that "We absolutely cannot relax our guard against ecological and environmental security problems sparked by the Three Gorges Project" (Bristow, 2007; McCabe, 2007). The question therefore was posed: how sustainable is the Three Gorges Project? Conventional approaches to sustainability assessment tend to use monetary based assessment aligned to triple bottom line thinking. That is, projects are evaluated as trade-offs between economic, environmental and social costs and benefits. The question of sustainability is considered using such a traditional Cost-Benefit Analysis approach, as undertaken in 1988 by a CIPM-Yangtze Joint Venture, and the Mauri Model Decision Making Framework (MMDMF). The Mauri Model differs from other approaches in that sustainability performance indicators are considered independently from any particular stakeholder bias. Bias is then introduced subsequently as a sensitivity analysis on the raw results obtained. The MMDMF is unique in that it is based on the Māori concept of Mauri, the binding force between the physical and the spiritual attributes of something, or the capacity to support life in the air, soil, and water. This concept of Mauri is analogous to the Chinese concept of Qi, and there are many analogous concepts in other cultures. It is the universal relevance of Mauri that allows its use to assess sustainability. This research identified that the MMDMF was a strong complement to Cost-Benefit Analysis, which is not designed as a sustainability assessment tool in itself. The

  5. The Origin of The Acheulean: The 1.7 Million-Year-Old Site of FLK West, Olduvai Gorge (Tanzania)

    PubMed Central

    Diez-Martín, F.; Sánchez Yustos, P.; Uribelarrea, D.; Baquedano, E.; Mark, D. F.; Mabulla, A.; Fraile, C.; Duque, J.; Díaz, I.; Pérez-González, A.; Yravedra, J.; Egeland, C. P.; Organista, E.; Domínguez-Rodrigo, M.

    2015-01-01

    The appearance of the Acheulean is one of the hallmarks of human evolution. It represents the emergence of a complex behavior, expressed in the recurrent manufacture of large-sized tools, with standardized forms, implying more advance forethought and planning by hominins than those required by the precedent Oldowan technology. The earliest known evidence of this technology dates back to c. 1.7 Ma. and is limited to two sites (Kokiselei [Kenya] and Konso [Ethiopia]), both of which lack functionally-associated fauna. The functionality of these earliest Acheulean assemblages remains unknown. Here we present the discovery of another early Acheulean site also dating to c. 1.7 Ma from Olduvai Gorge. This site provides evidence of the earliest steps in developing the Acheulean technology and is the oldest Acheulean site in which stone tools occur spatially and functionally associated with the exploitation of fauna. Simple and elaborate large-cutting tools (LCT) and bifacial handaxes co-exist at FLK West, showing that complex cognition was present from the earliest stages of the Acheulean. Here we provide a detailed technological study and evidence of the use of these tools on the butchery and consumption of fauna, probably by early Homo erectus sensu lato. PMID:26639785

  6. The binding sites of inhibitory monoclonal antibodies on acetylcholinesterase. Identification of a novel regulatory site at the putative "back door".

    PubMed

    Simon, S; Le Goff, A; Frobert, Y; Grassi, J; Massoulié, J

    1999-09-24

    We investigated the target sites of three inhibitory monoclonal antibodies on Electrophorus acetylcholinesterase (AChE). Previous studies showed that Elec-403 and Elec-410 are directed to overlapping but distinct epitopes in the peripheral site, at the entrance of the catalytic gorge, whereas Elec-408 binds to a different region. Using Electrophorus/rat AChE chimeras, we identified surface residues that differed between sensitive and insensitive AChEs: the replacement of a single Electrophorus residue by its rat homolog was able to abolish binding and inhibition, for each antibody. Reciprocally, binding and inhibition by Elec-403 and by Elec-410 could be conferred to rat AChE by the reverse mutation. Elec-410 appears to bind to one side of the active gorge, whereas Elec-403 covers its opening, explaining why the AChE-Elec-410 complex reacts faster than the AChE-Elec-403 or AChE-fasciculin complexes with two active site inhibitors, m-(N,N, N-trimethyltammonio)trifluoro-acetophenone and echothiophate. Elec-408 binds to the region of the putative "back door," distant from the peripheral site, and does not interfere with the access of inhibitors to the active site. The binding of an antibody to this novel regulatory site may inhibit the enzyme by blocking the back door or by inducing a conformational distortion within the active site.

  7. 6. VIEW FROM THE ROOF OF GORGE POWERHOUSE LOOKING EAST ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. VIEW FROM THE ROOF OF GORGE POWERHOUSE LOOKING EAST TO THE FORMER GRAVITY OIL STORAGE BUILDING, 1989. - Skagit Power Development, Gorge Powerhouse, On Skagit River, 0.4 mile upstream from Newhalem, Newhalem, Whatcom County, WA

  8. Raptor Use of the Rio Grande Gorge

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ponton, David A.

    2015-03-20

    The Rio Grande Gorge is a 115 km long river canyon located in Southern Colorado (15 km) and Northern New Mexico (100 km). The majority of the canyon is under the administration of the Bureau of Land Management {BLM), and 77 km of the canyon south of the Colorado/New Mexico border are designated Wild River under the National Wild and Scenic Rivers Act of 1968. Visits I have made to the Rio Grande Gorge over the past 15 .years disclosed some raptor utilization. As the Snake River Birds of Prey Natural Area gained publicity, its similarity to the Rio Grandemore » Gorge became obvious, and I was intrigued by the possibility of a high raptor nesting density in the Gorge. A survey in 1979 of 20 km of the northern end of the canyon revealed a moderately high density of red-tailed hawks and prairie falcons. With the encouragement of that partial survey, and a need to assess the impact of river-running on nesting birds of prey, I made a more comprehensive survey in 1980. The results of my surveys, along with those of a 1978 helicopter survey by the BLM, are presented in this report, as well as general characterization of the area, winter use by raptors, and an assessment of factors influencing the raptor population.« less

  9. Historic interior view of gorge wall, taken shortly after battle, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Historic interior view of gorge wall, taken shortly after battle, looking northwest at the entrance and northwest shoeing damage to gorge wall as well as timber and earth blindage (see also HABS No. GA-2158-39). - Fort Pulaski, Cockspur Island, Savannah, Chatham County, GA

  10. Unmet Expectations and Symptoms of Depression among the Three Gorges Project Resettlers

    PubMed Central

    Xi, Juan; Hwang, Sean-Shong

    2010-01-01

    To successfully resettle 1.27 million Chinese for the construction of the Three Gorges Project (TGP), the Chinese government employed a new re-resettlement policy which emphasizes infrastructural development of the resettlement sites to assure resettlers a sustainable livelihood following resettlement. Unfortunately, many benefits the policy promised have not materialized. As a result, many resettlers have suffered an increase in depressive symptoms associated with unmet expectations. Using panel data collected before and after relocation from a sample of Three Gorges resettlers, we found that a high proportion of resettlers reported unmet expectations which were significantly associated with an increase in depressive symptoms. Our findings indicate that undeliverable promises can backfire and further aggravate the harm inflicted on the displaced caused by the project-induced displacement. From the perspective of resettlers, a sound coping strategy to minimize the harm caused by the displacement on their mental wellbeing is to expect for the worst. PMID:21278844

  11. How a stationary knickpoint is sustained: New insights into the formation of the deep Yarlung Tsangpo Gorge

    NASA Astrophysics Data System (ADS)

    Wang, Yizhou; Zhang, Huiping; Zheng, Dewen; von Dassow, Wesley; Zhang, Zhuqi; Yu, Jingxing; Pang, Jianzhang

    2017-05-01

    In order to test the hypothesis that the stationary nature of the Yarlung Tsangpo Gorge is tectonically controlled, the rock uplift pattern in the southeast Tibetan Plateau and the critical condition to sustain a stable knickpoint must be derived. Via slope-area analysis and the integral approach, we first quantify the pattern of channel steepness in southeast Tibet and find that the steepness index shows higher values around the gorge but lower values toward the inner land and the mountain front. Such a pattern of channel steepness indicates that the active rock uplift is restricted in the zone just around the Yarlung Tsangpo Gorge. Then, we derive a general knickpoint migration model that accounts for spatially variant rock uplift rates. From the model, a critical condition for maintaining a stable knickpoint is concluded that the difference of incision rates in the downstream and upstream reaches of the knickpoint should match that of rock uplift. Employing a stream-power river incision model, we calculate the incision rate in the gorge and find a higher correspondence with differential rock uplift rates in the downstream and upstream reaches of the knickpoint. Therefore, we favor tectonic control as the primary mechanism to explain the stability of the knickpoint within the Yarlung Tsangpo Gorge.

  12. GEOMORPHOLOGY. Comment on "Tectonic control of Yarlung Tsangpo Gorge revealed by a buried canyon in Southern Tibet".

    PubMed

    Zeitler, Peter K; Koons, Peter O; Hallet, Bernard; Meltzer, Anne S

    2015-08-21

    Wang et al. (Reports, 21 November, 2014, p. 978) describe a buried canyon upstream of the Yarlung Tsangpo Gorge and argue that rapid erosion of the gorge was merely a passive response to rapid uplift at ~2.5 million years ago (Ma). We view these data as an expected consequence emerging from feedbacks between erosion and crustal rheology active well before 2.5 Ma. Copyright © 2015, American Association for the Advancement of Science.

  13. PAH distribution and mass fluxes in the Three Gorges Reservoir after impoundment of the Three Gorges Dam.

    PubMed

    Deyerling, Dominik; Wang, Jingxian; Hu, Wei; Westrich, Bernhard; Peng, Chengrong; Bi, Yonghong; Henkelmann, Bernhard; Schramm, Karl-Werner

    2014-09-01

    Mass fluxes of polycyclic aromatic hydrocarbons (PAHs) were calculated for the Three Gorges Reservoir (TGR) in China, based on concentration and discharge data from the Yangtze River. Virtual Organisms (VOs) have been applied during four campaigns in 2008, 2009 (twice) and 2011 at sampling sites distributed from Chongqing to Maoping. The total PAH mass fluxes ranged from 110 to 2,160 mg s(-1). Highest loads were determined at Chongqing with a decreasing trend towards Maoping in all four sampling campaigns. PAH remediation capacity of the TGR was found to be high as the mass flux reduced by more than half from upstream to downstream. Responsible processes are thought to be adsorption of PAH to suspended particles, dilution and degradation. Furthermore, the dependence of PAH concentration upon water depth was investigated at Maoping in front of the Three Gorges Dam. Although considerable differences could be revealed, there was no trend observable. Sampling of water with self-packed filter cartridges confirmed more homogenous PAH depth distribution. Moreover, PAH content of suspended particles was estimated from water concentrations gathered by VOs based on a water-particle separation model and subsequently compared to PAH concentration measured in water and in filter cartridges. It could be shown that the modeled data predicts the concentration caused by particle-bound PAHs to be about 6 times lower than PAHs dissolved in water. Besides, the model estimates the proportions of 5- and 6-ring PAHs being higher than in water phase. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. 20. MANUAL JACKING STATION UNIT 23 GORGE POWERHOUSE. JACKING FOR ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    20. MANUAL JACKING STATION UNIT 23 GORGE POWERHOUSE. JACKING FOR UNITS 23, 22, AND 21 HAS BEEN AUTOMATED FOR MANY YEARS BUT THE MANUAL JACKING STATIONS REMAIN IN PLACE AND FUNCTIONAL, 1989. - Skagit Power Development, Gorge Powerhouse, On Skagit River, 0.4 mile upstream from Newhalem, Newhalem, Whatcom County, WA

  15. Spatial patterns of erosion in a bedrock gorge

    NASA Astrophysics Data System (ADS)

    Beer, Alexander. R.; Turowski, Jens M.; Kirchner, James W.

    2017-01-01

    Understanding the physical processes driving bedrock channel formation is essential for interpreting and predicting the evolution of mountain landscapes. Here we analyze bedrock erosion patterns measured at unprecedented spatial resolution (mm) over 2 years in a natural bedrock gorge. These spatial patterns show that local bedrock erosion rates depend on position in the channel cross section, height above the streambed, and orientation relative to the main streamflow and sediment path. These observations are consistent with the expected spatial distribution of impacting particles (the tools effect) and shielding by sediment on the bed (the cover effect). Vertical incision by bedrock abrasion averaged 1.5 mm/a, lateral abrasion averaged 0.4 mm/a, and downstream directed abrasion of flow obstacles averaged 2.6 mm/a. However, a single plucking event locally exceeded these rates by orders of magnitude (˜100 mm/a), and accounted for one third of the eroded volume in the studied gorge section over the 2 year study period. Hence, if plucking is spatially more frequent than we observed in this study period, it may contribute substantially to long-term erosion rates, even in the relatively massive bedrock at our study site. Our observations demonstrate the importance of bedrock channel morphology and the spatial distribution of moving and static sediment in determining local erosion rates.

  16. Atmospheric deposition inputs and effects on lichen chemistry and indicator species in the Columbia River Gorge, USA.

    PubMed

    Fenn, M E; Geiser, L; Bachman, R; Blubaugh, T J; Bytnerowicz, A

    2007-03-01

    Topographic and meteorological conditions make the Columbia River Gorge (CRG) an 'exhaust pipe' for air pollutants generated by the Portland-Vancouver metropolis and Columbia Basin. We sampled fog, bulk precipitation, throughfall, airborne particulates, lichen thalli, and nitrophytic lichen distribution. Throughfall N and S deposition were high, 11.5-25.4 and 3.4-6.7 kg ha(-1) over 4.5 months at all 9 and 4/9 sites, respectively. Deposition and lichen thallus N were highest at eastern- and western-most sites, implicating both agricultural and urban sources. Fog and precipitation pH were frequently as low as 3.7-5.0. Peak NO(x), NH(3), and SO(2) concentrations in the eastern CRG were low, suggesting enhanced N and S inputs were largely from particulate deposition. Lichens indicating nitrogen-enriched environments were abundant and lichen N and S concentrations were 2x higher in the CRG than surrounding national forests. The atmospheric deposition levels detected likely threaten Gorge ecosystems and cultural resources.

  17. 18. SHEAR PIN, UNIT 24 GORGE POWERHOUSE. THE WICKET GATES ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. SHEAR PIN, UNIT 24 GORGE POWERHOUSE. THE WICKET GATES ON THE TURBINE ARE EACH EQUIPPED WITH A SHEAR PIN AND OIL PRESSURE GAUGE. IF A GATE JAMS, THE PIN SMEARS AND THE CHANGE IN OIL PRESSURE TRIGGERS AN ALARM, 1989. - Skagit Power Development, Gorge Powerhouse, On Skagit River, 0.4 mile upstream from Newhalem, Newhalem, Whatcom County, WA

  18. Unbinding of fluorinated oxime drug from the AChE gorge in polarizable water: a well-tempered metadynamics study.

    PubMed

    Pathak, Arup Kumar; Bandyopadhyay, Tusar

    2017-02-15

    Despite the fact that fluorination makes a drug more lipophilic, the molecular level understanding of protein-fluorinated drug interactions is very poor. Due to their enhanced ability to penetrate the blood brain barrier, they are suitable for reactivation of organophosphorus inactivated acetylcholinesterase (AChE) in the central nervous system. We systematically studied the unbinding of fluorinated obidoxime (FOBI) and non-fluorinated obidoxime (OBI) from the active site gorge of the serine hydrolase AChE in mean field polarizable water by employing all atom molecular dynamics simulations. It is observed that the unbinding process is strongly influenced by cation-π, hydrogen bond (HB) and water bridge interactions. The FOBI drug interacts more strongly with the protein residues than OBI and this is also verified from quantum mechanical calculations. Distinct unbinding pathways for FOBI and OBI are observed as evident from the 1D and 2D potential of mean force of the unbinding profiles. The present study suggests that the FOBI drug is held more firmly in the gorge of AChE in comparison to OBI and may lead to higher reactivation efficiency of the inactivated enzyme.

  19. Gravity Anomalies and Isostasy Deduced From New Dense Gravimetry Around the Tsangpo Gorge, Tibet

    NASA Astrophysics Data System (ADS)

    Fu, Guangyu; She, Yawen

    2017-10-01

    We built the first dense gravity network including 107 stations around the Tsangpo Gorge, Tibet, one of the hardest places in the world to reach, and conducted a gravity and hybrid GPS observation campaign in 2016. We computed the Bouguer gravity anomalies (BGAs) and free-air gravity anomalies (FGAs) and increased the resolution of the FGAs by merging the in situ data with EIGEN-6C4 gravity model data. The BGAs around the Tsangpo Gorge are in general negative and gradually decrease from south (-360 mGal) to north (-480 mGal). They indicate a uniformly dipping Moho around the Tsangpo Gorge that sinks from south to north at an angle of 12°. We introduced a method to compute the vertical tectonic stress of the lithosphere, a quantitative expression of isostasy, using BGA and terrain data, and applied it to the area around the Tsangpo Gorge. We found that the lithosphere of the upstream of the Tsangpo Gorge is roughly in an isostatic state, but the lithosphere of the downstream exhibits vertical tectonic stress of 50 MPa, which indicates the loss of a large amount of surface material. This result does not support the deduction of the valley bottom before uplift of the Tsangpo Gorge by Wang et al. (2014).

  20. Recreation impacts to cliff resources in the Potomac Gorge: Final report, June 2011

    USGS Publications Warehouse

    Marion, Jeffrey L.; Carr, C.; Davis, C.A.

    2011-01-01

    Managers of the National Park Service (NPS) are directed by law to accommodate appropriate types and amounts of visitation while ensuring that: any adverse impacts are the minimum necessary, unavoidable, cannot be further mitigated, and do not constitute impairment or derogation of park resources and values. (NPS 2006). The increasing popularity of the national park system presents substantial management challenges. High visitatation may cause unacceptable impacts to fragile natural and cultural resources, and may also cause crowding and other social impacts, which can also degrade the quality of visitor experiences. Responding to these concerns, NPS managers at Chesapeake & Ohio Canal National Historical Park (CHOH) and George Washington Memorial Parkway (GWMP) sponsored this research within the upper Potomac Gorge portions of these parks to investigate visitation-related impacts to the park?s cliff resources. The cliffs and rocky areas within the Great Falls and Mather Gorge areas provide important habitats for numerous sensitive rare plants and plant communities. A recent General Management Planning process for Great Falls Park (GFP), a portion of GWMP, highlighted the potential impacts of cliff-associated recreational activities, including hiking, climbing, and fishing, on sensitive cliff resources. The planning process identified the need for development of a Climbing Management Plan and a Trail Plan to more specifically address site and visitor management actions needed to protect rare and sensitive natural and cultural resources. Good science to assess cliff-associated rare plants and communities and to determine the existing and potential effects of cliff-related recreational activities is required for these new planning efforts. This research is designed to specifically address these informational needs and to assist park managers on both sides of the river with current and future cliff and recreation management decisions.

  1. Spatial forecast of landslides in three gorges based on spatial data mining.

    PubMed

    Wang, Xianmin; Niu, Ruiqing

    2009-01-01

    The Three Gorges is a region with a very high landslide distribution density and a concentrated population. In Three Gorges there are often landslide disasters, and the potential risk of landslides is tremendous. In this paper, focusing on Three Gorges, which has a complicated landform, spatial forecasting of landslides is studied by establishing 20 forecast factors (spectra, texture, vegetation coverage, water level of reservoir, slope structure, engineering rock group, elevation, slope, aspect, etc). China-Brazil Earth Resources Satellite (Cbers) images were adopted based on C4.5 decision tree to mine spatial forecast landslide criteria in Guojiaba Town (Zhigui County) in Three Gorges and based on this knowledge, perform intelligent spatial landslide forecasts for Guojiaba Town. All landslides lie in the dangerous and unstable regions, so the forecast result is good. The method proposed in the paper is compared with seven other methods: IsoData, K-Means, Mahalanobis Distance, Maximum Likelihood, Minimum Distance, Parallelepiped and Information Content Model. The experimental results show that the method proposed in this paper has a high forecast precision, noticeably higher than that of the other seven methods.

  2. Columbia Gorge Community College Business Survey.

    ERIC Educational Resources Information Center

    McKee, Jonathon V.

    This is a report on a business survey conducted by Columbia Gorge Community College (CGCC) (Oregon) to review the success and quality of the college's degree and certificate programs in business administration, computer application systems, and computer information systems. The community college surveyed 104 local businesses to verify the…

  3. When felids and hominins ruled at Olduvai Gorge: A machine learning analysis of the skeletal profiles of the non-anthropogenic Bed I sites

    NASA Astrophysics Data System (ADS)

    Arriaza, Mari Carmen; Domínguez-Rodrigo, Manuel

    2016-05-01

    In the past twenty years, skeletal part profiles, which are prone to equifinality, have not occupied a prominent role in the interpretation of early Pleistocene sites on Africa. Alternatively, taphonomic studies on bone surface modifications and bone breakage patterns, have provided heuristic interpretations of some of the best preserved archaeological record of this period; namely, the Olduvai Bed I sites. The most recent and comprehensive taphonomic study of these sites (Domínguez-Rodrigo et al., 2007a) showed that FLK Zinj was an anthropogenic assemblage in which hominins acquired carcasses via primary access. That study also showed that the other sites were palimpsests with minimal or no intervention by hominins. The FLK N, FLK NN and DK sequence seemed to be dominated by single-agent (mostly, felid) or multiple-agent (mostly, felid-hyenid) processes. The present study re-analyzes the Bed I sites focusing on skeletal part profiles. Machine learning methods, which incorporate complex algorithms, are powerful predictive and classification methods and have the potential to better extract information from skeletal part representation than past approaches. Here, multiple algorithms (via decision trees, neural networks, random forests and support vector machines) are combined to produce a solid interpretation of bone accumulation agency at the Olduvai Bed I sites. This new approach virtually coincides with previous taphonomic interpretations on a site by site basis and shows that felids were dominant accumulating agents over hyenas during Bed I times. The recent discovery of possibly a modern lion-accumulated assemblage at Olduvai Gorge (Arriaza et al., submitted) provides a very timely analog for this interpretation.

  4. Spatial Forecast of Landslides in Three Gorges Based On Spatial Data Mining

    PubMed Central

    Wang, Xianmin; Niu, Ruiqing

    2009-01-01

    The Three Gorges is a region with a very high landslide distribution density and a concentrated population. In Three Gorges there are often landslide disasters, and the potential risk of landslides is tremendous. In this paper, focusing on Three Gorges, which has a complicated landform, spatial forecasting of landslides is studied by establishing 20 forecast factors (spectra, texture, vegetation coverage, water level of reservoir, slope structure, engineering rock group, elevation, slope, aspect, etc). China-Brazil Earth Resources Satellite (Cbers) images were adopted based on C4.5 decision tree to mine spatial forecast landslide criteria in Guojiaba Town (Zhigui County) in Three Gorges and based on this knowledge, perform intelligent spatial landslide forecasts for Guojiaba Town. All landslides lie in the dangerous and unstable regions, so the forecast result is good. The method proposed in the paper is compared with seven other methods: IsoData, K-Means, Mahalanobis Distance, Maximum Likelihood, Minimum Distance, Parallelepiped and Information Content Model. The experimental results show that the method proposed in this paper has a high forecast precision, noticeably higher than that of the other seven methods. PMID:22573999

  5. Canyons and gorges as potential geotourism destinations in Serbia: comparative analysis from two perspectives - general geotourists' and pure geotourists'

    NASA Astrophysics Data System (ADS)

    Božić, Sanja; Tomić, Nemanja

    2015-10-01

    Serbia represents one of those countries which have not yet differentiated themselves on the world tourism map. However, it has an immense but still unrevealed potential for geotourism development. In this paper we analyzed several remarkable canyons and gorges of great scientific importance and geotourism potential. These sites include the Djerdap Gorge and Lazar River Canyon, located in Eastern Serbia and the Ovcar-Kablar Gorge and Uvac Canyon located in Western Serbia. One of the main goals of this paper was to analyze and compare the current state and tourism potential of these geosites by using the M-GAM model for geosite assessment. However, the principal aim of the paper is to analyze how important is each subindicator in the assessment process for different market segments. In this paper, we also analyzed how giving different importance to subindicators can influence the position of the geosites in the matrix indicating different assessment done by two chosen market segments. The research showed that general geotourists appreciate considerably different values when assessing a geosite in comparison to pure geotourists. The paper can be used as framework for developing the tourism management strategy of geosites taking into consideration the needs and preferences of the target market segments.

  6. Linking Ah receptor mediated effects of sediments and impacts on fish to key pollutants in the Yangtze Three Gorges Reservoir, China - A comprehensive perspective.

    PubMed

    Floehr, Tilman; Scholz-Starke, Björn; Xiao, Hongxia; Hercht, Hendrik; Wu, Lingling; Hou, Junli; Schmidt-Posthaus, Heike; Segner, Helmut; Kammann, Ulrike; Yuan, Xingzhong; Roß-Nickoll, Martina; Schäffer, Andreas; Hollert, Henner

    2015-12-15

    The Three Gorges Reservoir (TGR), created in consequence of the Yangtze River's impoundment by the Three Gorges Dam, faces numerous anthropogenic impacts that challenge its unique ecosystem. Organic pollutants, particularly aryl hydrocarbon receptor (AhR) agonists, have been widely detected in the Yangtze River, but only little research was yet done on AhR-mediated activities. Hence, in order to assess effects of organic pollution, with particular focus on AhR-mediated activities, several sites in the TGR area were examined applying the "triad approach". It combines chemical analysis, in vitro, in vivo and in situ investigations to a holistic assessment. Sediments and the benthic fish species Pelteobagrus vachellii were sampled in 2011/2012, respectively, to identify relevant endpoints. Sediment was tested in vitro with the ethoxyresorufin-O-deethylase (EROD) induction assay, and in vivo with the Fish Embryo Toxicity Test and Sediment Contact Assay with Danio rerio. Activities of phase I (EROD) and phase II (glutathione-S-transferase) biotransformation enzymes, pollutant metabolites and histopathological alterations were studied in situ in P. vachellii. EROD induction was tested in vitro and in situ to evaluate possible relationships. Two sites, near Chongqing and Kaixian city, were identified as regional hot-spots and further investigated in 2013. The sediments induced in the in vitro/in vivo bioassays AhR-mediated activities and embryotoxic/teratogenic effects - particularly on the cardiovascular system. These endpoints could be significantly correlated to each other and respective chemical data. However, particle-bound pollutants showed only low bioavailability. The in situ investigations suggested a rather poor condition of P. vachellii, with histopathological alterations in liver and excretory kidney. Fish from Chongqing city exhibited significant hepatic EROD induction and obvious parasitic infestations. The polycyclic aromatic hydrocarbon (PAH) metabolite 1

  7. The Volcanic Story of the Columbia River Gorge.

    ERIC Educational Resources Information Center

    Allen, John Eliot

    1982-01-01

    Uses maps, diagrams, and geological accounts to describe the Columbia River Gorge which separates Oregon from Washington and exposes an unexcelled record of Cenozoic volcanic stratigraphy, sedimentation, flooding, and landsliding for the visiting tourists, geologists, or students. (Author/DC)

  8. Rapid Gorge Formation in an Artificially Created Waterfall

    NASA Astrophysics Data System (ADS)

    Anton, L.; Mather, A. E.; Stokes, M.; Munoz Martin, A.

    2014-12-01

    A number of studies have examined rates of gorge formation, nick point retreat, and the controls on those rates via bedrock erodibility, the effectiveness of bedrock erosion mechanisms and the role of hillslope processes. Most findings are based on conceptual / empirical models or long term landscape analysis; but studies of recent quantifiable events are scarce yet highly valuable. Here we present expert eye witness account and quantitative survey of large and rapid fluvial erosion events that occurred over an artificially created waterfall at a spillway mouth. In 6 years a ~270 m long, ~100 m deep and ~100 to 160 m wide canyon was carved, and ~1.58 x106 m3 of granite bedrock was removed from the spillway site. Available flow data indicates that the erosion took place under unremarkable flood discharge conditions. The analysis of historic topographic maps enables the reconstruction of the former topography and successive erosion events, enabling the quantification of bedrock erosion amounts, and rates. Analysis of bedrock erodibility and discontinuity patterns demonstrates that the bedrock is mechanically strong, and that similar rock strength and fracture patterns are found throughout the region. It is apparent that structural pre-conditioning through fracture density and orientation in relation to flow and slope direction is of paramount importance in the gorge development. The presented example provides an exceptional opportunity for studying the evolution process of a bedrock canyon and to precisely measure the rate of bedrock channel erosion over a six year period. Results illustrate the highly episodic nature of the erosion and highlight several key observations for the adjustability of bedrock rivers. The observations have implications for the efficiency of bedrock erosion and raise important questions about incision rates, driving mechanisms and timescale assumptions' in models of landscape change.

  9. Water Pollution Prediction in the Three Gorges Reservoir Area and Countermeasures for Sustainable Development of the Water Environment.

    PubMed

    Li, Yinghui; Huang, Shuaijin; Qu, Xuexin

    2017-10-27

    The Three Gorges Project was implemented in 1994 to promote sustainable water resource use and development of the water environment in the Three Gorges Reservoir Area (hereafter "Reservoir Area"). However, massive discharge of wastewater along the river threatens these goals; therefore, this study employs a grey prediction model (GM) to predict the annual emissions of primary pollution sources, including industrial wastewater, domestic wastewater, and oily and domestic wastewater from ships, that influence the Three Gorges Reservoir Area water environment. First, we optimize the initial values of a traditional GM (1,1) model, and build a new GM (1,1) model that minimizes the sum of squares of the relative simulation errors. Second, we use the new GM (1,1) model to simulate historical annual emissions data for the four pollution sources and thereby test the effectiveness of the model. Third, we predict the annual emissions of the four pollution sources in the Three Gorges Reservoir Area for a future period. The prediction results reveal the annual emission trends for the major wastewater types, and indicate the primary sources of water pollution in the Three Gorges Reservoir Area. Based on our predictions, we suggest several countermeasures against water pollution and towards the sustainable development of the water environment in the Three Gorges Reservoir Area.

  10. Monitoring the Dynamic of a Fluvial Channel after Lahar Disturbance: Huiloac Gorge (Popocatepetl Volcano, Mexico)

    NASA Astrophysics Data System (ADS)

    Andres, N.; Palacios, D.; Zamorano, J. J.; Tanarro, L. M.; Renschler, C.; Sanjosé, J. J.; Atkinson, A.

    2009-04-01

    Volcanic eruptions generate disturbances that affect hydrological systems (Major, 2003) by depositing large volumes of sediments in watersheds that exceed amounts common to non-volcanic river systems (Montgomery, 2005). If the eruption releases abundant melt water, the river system may respond immediately by forming hazardous flows called lahars. River system recovery following eruptive and laharic impact is an important process, but it has received little attention (Gran and Montgomery, 2005) despite the fact that Major et al. (2000) and Hayes et al. (2002) have shown that these disruptions cause long term instability and their effects persist for decades. Lahar deposits resulting from interaction between volcanic activity and the glacier located above the Huiloac Gorge on the northern slope of Popocatepetl volcano (19°02´ N, 98°62´ W, 5,424 m), have infilled the gorge (Palacios, 1995; Palacios et al., 1998 and 2001; Capra et al., 2004; Muñoz, 2007). All of the major lahars that occurred on the volcano in 1995 (4 km), 1997 (21 km), and 2001 (14 km) have channelled through Huiloac Gorge, and have dramatically altered its morphology and dynamics through erosion and deposition. The present study traces these changes in the aftermath of the laharic events that occurred from 1997-2001. A sector of the channel, located at 3200m-3240m altitude, of 500 m long and 15 to 20 m wide, in the mid-section of the gorge, was chosen as the control site. Precipitation is heaviest there and is most apt to trigger secondary post-eruptive lahars. ArcGis software was used to draw 6 geomorphic maps of the site showing spatial variations in the landforms for the period February 2002 - February 2008. In addition, 29 cross-profiles were made of the gorge for the same time interval, excluding February 2004. The volume of sediment eroded and deposited was calculated for each date by comparing variations in the height of the floor and banks of the gorge depicted in the cross-profile, and

  11. Unraveling hominin behavior at another anthropogenic site from Olduvai Gorge (Tanzania): new archaeological and taphonomic research at BK, Upper Bed II.

    PubMed

    Domínguez-Rodrigo, M; Mabulla, A; Bunn, H T; Barba, R; Diez-Martín, F; Egeland, C P; Espílez, E; Egeland, A; Yravedra, J; Sánchez, P

    2009-09-01

    New archaeological excavations and research at BK, Upper Bed II (Olduvai Gorge, Tanzania) have yielded a rich and unbiased collection of fossil bones. These new excavations show that BK is a stratified deposit formed in a riverine setting close to an alluvial plain. The present taphonomic study reveals the second-largest collection of hominin-modified bones from Olduvai, with abundant cut marks found on most of the anatomical areas preserved. Meat and marrow exploitation is reconstructed using the taphonomic signatures left on the bones by hominins. Highly cut-marked long limb shafts, especially those of upper limb bones, suggest that hominins at BK were actively engaged in acquiring small and middle-sized animals using strategies other than passive scavenging. The exploitation of large-sized game (Pelorovis) by Lower Pleistocene hominins, as suggested by previous researchers, is supported by the present study.

  12. 78 FR 73092 - Special Regulations; Areas of the National Park System, New River Gorge National River, Bicycling

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-05

    ... recreation and tourism activities in southern West Virginia due to its nationally recognized status and has... for tourism and a catalyst for economic development in the New River Gorge area, as it attracts... through a commitment to consultation with Indian tribes and recognition of their right to self-governance...

  13. Water Pollution Prediction in the Three Gorges Reservoir Area and Countermeasures for Sustainable Development of the Water Environment

    PubMed Central

    Huang, Shuaijin; Qu, Xuexin

    2017-01-01

    The Three Gorges Project was implemented in 1994 to promote sustainable water resource use and development of the water environment in the Three Gorges Reservoir Area (hereafter “Reservoir Area”). However, massive discharge of wastewater along the river threatens these goals; therefore, this study employs a grey prediction model (GM) to predict the annual emissions of primary pollution sources, including industrial wastewater, domestic wastewater, and oily and domestic wastewater from ships, that influence the Three Gorges Reservoir Area water environment. First, we optimize the initial values of a traditional GM (1,1) model, and build a new GM (1,1) model that minimizes the sum of squares of the relative simulation errors. Second, we use the new GM (1,1) model to simulate historical annual emissions data for the four pollution sources and thereby test the effectiveness of the model. Third, we predict the annual emissions of the four pollution sources in the Three Gorges Reservoir Area for a future period. The prediction results reveal the annual emission trends for the major wastewater types, and indicate the primary sources of water pollution in the Three Gorges Reservoir Area. Based on our predictions, we suggest several countermeasures against water pollution and towards the sustainable development of the water environment in the Three Gorges Reservoir Area. PMID:29077006

  14. After Three Gorges Dam: What have we learned?

    NASA Astrophysics Data System (ADS)

    Natali, J.; Williams, P.; Wong, R.; Kondolf, G. M.

    2013-12-01

    China is at a critical point in its development path. By investing heavily in large-scale infrastructure, the rewards of economic growth weigh against long-term environmental and social costs. The construction of Three Gorges Dam, the world's largest hydroelectric project, began in 1994. Between 2002 and 2010, its 660 kilometer reservoir filled behind a 181 meter dam, displacing at least 1.4 million people and transforming Asia's longest river (the Yangtze) while generating nearly 100 billion kWh/yr of electricity -- 2.85% of China's current electric power usage. As the mega-project progenitor in a cascade of planned dams, the Three Gorges Dam emerges as a test case for how China will plan, execute and mitigate its development pathway and the transformation of its environment. Post-Project Assessments (PPA) provide a systematic, scientific method for improving the practice of environmental management - particularly as they apply to human intervention in river systems. In 2012, the Department of Landscape Architecture and Environmental Planning at University of California, Berkeley organized a symposium-based PPA for the Three Gorges Dam on the Yangtze River. Prior to this symposium, the twelve invited Chinese scientists, engineers and economists with recent research on Three Gorges Dam had not had the opportunity to present their evaluations together in an open, public forum. With a 50-year planning horizon, the symposium's five sessions centered on impacts on flows, geomorphology, geologic hazards, the environment and socioeconomic effects. Three Gorges' project goals focused on flood control, hydropower and improved navigation. According to expert research, major changes in sediment budget and flow regime from reservoir operation have significantly reduced sediment discharge into the downstream river and estuary, initiating a series of geomorphic changes with ecological and social impacts. While the dam reduces high flow stages from floods originating above the

  15. Temperature changes in Three Gorges Reservoir Area and linkage with Three Gorges Project

    NASA Astrophysics Data System (ADS)

    Song, Zhen; Liang, Shunlin; Feng, Lian; He, Tao; Song, Xiao-Peng; Zhang, Lei

    2017-05-01

    The Three Gorges Project (TGP) is one of the largest hydroelectric projects throughout the world. It has brought many benefits to the society but also led to endless debates about its environmental and climatic impacts. Monitoring the spatiotemporal variations of temperature in the Three Gorges Reservoir Area (TGRA) is important for understanding the climatic impacts of the TGP. In this study, we used remote sensing-based land surface temperature (LST) and ground-measured air temperature data to investigate temperature changes in the TGRA. Results showed that during the daytime in summer, LST exhibited significant cooling (1-5°C) in the downstream region of the reservoir, whereas LST during the nighttime in winter exhibited significant warming (1-5°C) across the entire reservoir. However, these cooling and warming effects were both locally constrained within 5 km buffer along the reservoir. The changes in air temperature were consistent with those in LST, with 0.67°C cooling in summer and 0.33°C warming in winter. The temperature changes along the reservoir not only resulted from the land-water conversion induced by the dam impounding but were also related to the increase of vegetation cover caused by the ecological restoration projects. Significant warming trends were also found in the upstream of TGRA, especially during the daytime in summer, with up to 5°C for LST and 0.52°C for air temperature. The warming was caused mainly by urban expansion, which was driven in part by the population resettlement of TGP. Based on satellite observations, we investigated the comprehensive climatic impacts of TGP caused by multiple factors.

  16. Assessing internal biophysical vulnerability to landslide hazards - a nested catchment approach: Xiangxi Watershed / Three Gorges Reservoir

    NASA Astrophysics Data System (ADS)

    Wiegand, Matthias; Seeber, Christoph; Hartmann, Heike; Xiang, Wei; King, Lorenz

    2010-05-01

    The Three Gorges dam construction was completed in 2006. Besides the international media, also the responsible authorities and various scholarly communities pay close attention to potential and actual environmental impacts related to the impoundment and development activities. The geo-environment within the Three Gorges region is highly conducive to landslides. Consequently, a scientific monitoring and risk mitigation system was established and is still under development. Risk analysis with regard to gravity driven mass movements is highly complex and strongly site specific - several aspects hamper a universal methodology applicable for landslide risk and site assessment. The interdisciplinary Sino-German Yangtze-Project Research co-operation aims, among others, to support the sustainable cultivation of the newly developed ecosystems within the Yangtze catchments. Land use change and increasing population growth are causing severe pressure on the scarce land resources. Landslides are acknowledged as important threat, hence vulnerability of certain landscape components have to be identified, quantified and monitored. A nested quantitative approach for vulnerability analysis is developed. The applied risk and vulnerability model understands risk as the product of hazard and vulnerability. Whereas vulnerability is characterized by: mass movement intensity and susceptibility of the respective element at risk. The watershed of Xiangxi river serves as study area. In general, catchment approaches intent and proved to be a functional geographical unit for successful integrated resources management. Several limitations with regard to data accessibility, availability and accuracy have to be considered due to restrictions of feasible scales. Comprehensive large-scale site investigations are confined to training areas for model calibration and validation. Remote sensing potentials are utilised for land use/ land cover change analysis and localization of selected elements

  17. Short-term food restriction followed by controlled refeeding promotes gorging behavior, enhances fat deposition, and diminishes insulin sensitivity in mice

    PubMed Central

    Kliewer, Kara L.; Ke, Jia-Yu; Stout, Michael B.; Cole, Rachel; Samuel, Varman T.; Shulman, Gerald I.; Belury, Martha A.

    2015-01-01

    Rodents are commonly used in food restriction-refeeding studies to investigate weight regain. Mice that are rationed food every 24 hours may consume all allocated food in a short time (gorge) and therefore undergo a brief well-fed period followed by an extended fasted period until the next day’s food allotment. These exaggerated metabolic states are not typical in ad-libitum fed (nibbling) mice. The aim of the current study was to elucidate the intraday and cumulative metabolic consequences of gorging (induced by food restriction) in mice during controlled refeeding. Accordingly, following a temporary food restriction, mice were fed rations similar to intakes of ad-libitum fed controls. Temporary food restriction initiated gorging behavior that persisted during refeeding; consequently, metabolism-related measurements were obtained in the gorging mice during their daily fed and fasted metabolic states. Robust differences in adipose tissue lipogenic and inflammatory gene expression were found in the gorging mice by metabolic state (fed versus fasted). Additionally, despite a reduced cumulative food intake compared to ad-libitum fed mice, restriction-induced gorging mice had increased intra-abdominal fat accumulation, diminished hepatic and peripheral insulin sensitivity, and a gene expression profile favoring lipid deposition. Our findings highlight the intraday differences in gene expression in gorging mice before and after feeding that confound comparisons with ad-libitum fed, or nibbling, mice. The present study also provides evidence that weight regain following food restriction is associated with cumulative metabolic and behavioral abnormalities in mice. PMID:25913018

  18. Geomorphic constraints on the evolution of the Kern Gorge, southern Sierra Nevada, California.

    NASA Astrophysics Data System (ADS)

    Foreshee, B. C.; Krugh, W. C.

    2016-12-01

    The Kern River is uniquely positioned to respond to tectonic activity that occurs within the southern Sierra Nevada and southern San Joaquin Basin, CA. The North and South forks of the upper Kern River (above Lake Isabella) are fed by tributaries that primarily drain the high-elevation low-relief landscape of the Kern Plateau. These south flowing trunk streams switch to a dominantly southwest flow direction at the Lake Isabella Reservoir and South Lake Valley respectively. Downstream from Lake Isabella, the Kern River steepens as it flows through the Kern Gorge and then crosses the Kern Arch region of the San Joaquin Basin. Clark et al., (2005) used low-temperature thermochronometry and trunk and tributary channel profiles from the upper Kern River catchment to identify two periods of rapid incision that occurred from 32.0 to 3.5 Ma and from 3.5 Ma to present. Cecil et al., (2014) used low-temperature thermochronometry from well cores of Oligocene-Miocene sandstones to investigate the time-temperature history of the Kern Arch and identified a period of subsidence and sedimentation between 6.0 and 1.0 Ma that was immediately followed by rapid exhumation. They attributed these results to the northwest migration of a delaminating lithospheric root. In this study we examine the erosional and depositional history within the Kern Gorge to investigate the response of the Kern River to Pliocene-Pleistocene tectonic activity within the Kern Arch and southern Sierra Nevada. Quantitative stream profile analyses and geomorphic mapping within the Kern Gorge are being conducted using USGS 10m DEM data, satellite and aerial imagery, and field based observations and measurements. Reconnaissance mapping efforts have so far identified several strath terraces, alluvial fill terraces, colluvial deposits, and multiple debris flow and landslide deposits that have been incised by the Kern River and are now preserved above the active channel. These geomorphic landforms are currently being

  19. Geologic map of Gunnison Gorge National Conservation Area, Delta and Montrose Counties, Colorado

    USGS Publications Warehouse

    Kellogg, Karl; Hansen, Wallace R.; Tucker, Karen S.; VanSistine, D. Paco

    2004-01-01

    This publication consists of a geologic map database and printed map sheet. The map sheet has a geologic map as the center piece, and accompanying text describes (1) the various geological units, (2) the uplift history of the region and how it relates to canyon downcutting, (3) the ecology of the gorge, and (4) human history. The map is intended to be used by the general public as well as scientists and goes hand-in-hand with a separate geological guide to Gunnison Gorge.

  20. Is there a Developed Oldowan A at Olduvai Gorge? A diachronic analysis of the Oldowan in Bed I and Lower-Middle Bed II at Olduvai Gorge, Tanzania.

    PubMed

    Proffitt, Tomos

    2018-05-08

    Debates regarding the validity of the Developed Oldowan as separate cultural facies within the Oldowan techno-complex have primarily concentrated on the Developed Oldowan B/Acheulean transition, with little attention paid to the validity of the Developed Oldowan A (DOA) as a valid technological differentiation. This study presents a diachronic technological analysis and comparison of Oldowan and DOA lithic assemblages from Olduvai Gorge, Tanzania, dated between 1.84 and 1.6 Ma, to test the validity of Leakey's original distinction between these two cultural facies. The results from this comparative analysis show very few technological differences between the lithic assemblages previously assigned to the DOA and Classic Oldowan. Significant diachronic variation in raw material availability and use is, however, identified between Bed I and Lower/Middle Bed II of Olduvai Gorge, which may go some way to explaining the originally perceived techno-cultural differences. The results suggest an increase in hominin knapping and percussive activities, as well as a clear ability to preferentially select high quality raw materials stratigraphically above Tuff IF. Technological innovation and complexity, however, does not seem to vary significantly between the Classic Oldowan and DOA assemblages. The results of this analysis along with similar studies from the wider eastern African region lead to the conclusion that the term Developed Oldowan A should no longer be used. Copyright © 2018 Elsevier Ltd. All rights reserved.

  1. The Project for Developing Countermeasures against Landslides in the Abay River Gorge, Ethiopia

    NASA Astrophysics Data System (ADS)

    Guta, H. E.

    2016-12-01

    The Blue Nile Gorge of Ethiopia is characterized by high relief landscape. The stretch of major arterial road that connects Ethiopia to Sudan passes through the Gorge. The Gorge is plagued by swarms of landslides which makes it a tremendous obstacle for travel and communication. Therefore, landside study was carried out from 2010 to 2012 by JICA and Geological survey of Ethiopia to figure out the mechanisms that trigger the swarms of landslides that occur in the area and identify appropriate countermeasures that would be best implemented. The study included geomorphologic and geologic survey, drilling survey, displacement monitoring, ground water and precipitation monitoring, geophysical exploration, and stability analysis. About 42 landslide monitoring instruments namely extensometers (both surface and borehole), inclinometers and ground water level meters were installed in four highly landslide prone areas to detect slip surface, and determine amount and direction of movement. The amount of landslide movement at the four zones is 42.4, 57.6, 294.9 and 136mm during rainy season. Ground water level rising, nature of material and intense rainfall are found to be among the major triggering factors. Stability analysis using Simple Jambu and modified Fellenus methods was conducted resulting in safety factor Fs less than one and reasonably 0.98 by adopting shear parameters of soils by back analysis. By assuming cohesion (c') to be very close to 0 due to landslide blocks active movement when ground water rises during rainy season, Shear resistance angle, ɸ, was obtained to be 10.80, 26.30, 10.20 and 16.30 in the four areas using Modified Fellenius method and 10.70, 26.60, 10.00 and 16.10 using Simple Janbu method. Effect of countermeasures was checked by trial calculation. Accordingly the factory of safety increased from 0.98 to 1.2 when ɸ=60, ground water is lowered by 6m, and steel pipe pile of ɸ500mm x t40mm at an interval of 1.9m are implemented. consequently

  2. Effects of viscosity and osmotic stress on the reaction of human butyrylcholinesterase with cresyl saligenin phosphate, a toxicant related to aerotoxic syndrome: kinetic and molecular dynamics studies.

    PubMed

    Masson, Patrick; Lushchekina, Sofya; Schopfer, Lawrence M; Lockridge, Oksana

    2013-09-15

    CSP (cresyl saligenin phosphate) is an irreversible inhibitor of human BChE (butyrylcholinesterase) that has been involved in the aerotoxic syndrome. Inhibition under pseudo-first-order conditions is biphasic, reflecting a slow equilibrium between two enzyme states E and E'. The elementary constants for CSP inhibition of wild-type BChE and D70G mutant were determined by studying the dependence of inhibition kinetics on viscosity and osmotic pressure. Glycerol and sucrose were used as viscosogens. Phosphorylation by CSP is sensitive to viscosity and is thus strongly diffusion-controlled (kon≈10⁸ M⁻¹·min⁻¹). Bimolecular rate constants (ki) are about equal to kon values, making CSP one of the fastest inhibitors of BChE. Sucrose caused osmotic stress because it is excluded from the active-site gorge. This depleted the active-site gorge of water. Osmotic activation volumes, determined from the dependence of ki on osmotic pressure, showed that water in the gorge of the D70G mutant is more easily depleted than that in wild-type BChE. This demonstrates the importance of the peripheral site residue Asp⁷⁰ in controlling the active-site gorge hydration. MD simulations provided new evidence for differences in the motion of water within the gorge of wild-type and D70G enzymes. The effect of viscosogens/osmolytes provided information on the slow equilibrium E⇌E', indicating that alteration in hydration of a key catalytic residue shifts the equilibrium towards E'. MD simulations showed that glycerol molecules that substitute for water molecules in the enzyme active-site gorge induce a conformational change in the catalytic triad residue His⁴³⁸, leading to the less reactive form E'.

  3. Accumulation of floating microplastics behind the Three Gorges Dam.

    PubMed

    Zhang, Kai; Gong, Wen; Lv, Jizhong; Xiong, Xiong; Wu, Chenxi

    2015-09-01

    We investigated the occurrence and distribution of microplastics in surface water from the Three Gorges Reservoir. Nine samples were collected via trawl sampling with a 112 μmmesh net. The abundances of microplastics were from 3407.7 × 10(3) to 13,617.5 × 10(3) items per square kilometer in the main stream of the Yangtze River and from 192.5 × 10(3) to 11,889.7 × 10(3) items per square kilometer in the estuarine areas of four tributaries. The abundance of microplastics in the main stream of the Yangtze River generally increased as moving closer to the Three Gorges Dam. The microplastics are made exclusively of polyethylene (PE), polypropylene (PP), and polystyrene (PS). Together with microplastics, high abundance of coal/fly ash was also observed in the surface water samples. Comparing with previously reported data, microplastics in the TGR were approximately one to three orders of magnitudes greater, suggesting reservoirs as potential hot spot for microplastic pollution. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. 33 CFR 110.127b - Flaming Gorge Lake, Wyoming-Utah.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... following points, excluding a 150-foot-wide fairway, extending southeasterly from the launching ramp, as... inclosed by the shore and a line connecting the following points, excluding a 100-foot-wide fairway.... (c) Antelope Flat, Utah. That portion of Flaming Gorge Lake inclosed by the shore and a line...

  5. Breeding systems, hybridization and continuing evolution in Avon Gorge Sorbus

    PubMed Central

    Ludwig, Shanna; Robertson, Ashley; Rich, Timothy C. G.; Djordjević, Milena; Cerović, Radosav; Houston, Libby; Harris, Stephen A.; Hiscock, Simon J.

    2013-01-01

    Background and Aims Interspecific hybridization and polyploidy are key processes in plant evolution and are responsible for ongoing genetic diversification in the genus Sorbus (Rosaceae). The Avon Gorge, Bristol, UK, is a world ‘hotspot’ for Sorbus diversity and home to diploid sexual species and polyploid apomictic species. This research investigated how mating system variation, hybridization and polyploidy interact to generate this biological diversity. Methods Mating systems of diploid, triploid and tetraploid Sorbus taxa were analysed using pollen tube growth and seed set assays from controlled pollinations, and parent–offspring genotyping of progeny from open and manual pollinations. Key Results Diploid Sorbus are outcrossing and self-incompatible (SI). Triploid taxa are pseudogamous apomicts and genetically invariable, but because they also display self-incompatibility, apomictic seed set requires pollen from other Sorbus taxa – a phenomenon which offers direct opportunities for hybridization. In contrast tetraploid taxa are pseudogamous but self-compatible, so do not have the same obligate requirement for intertaxon pollination. Conclusions The mating inter-relationships among Avon Gorge Sorbus taxa are complex and are the driving force for hybridization and ongoing genetic diversification. In particular, the presence of self-incompatibility in triploid pseudogamous apomicts imposes a requirement for interspecific cross-pollination, thereby facilitating continuing diversification and evolution through rare sexual hybridization events. This is the first report of naturally occurring pseudogamous apomictic SI plant populations, and we suggest that interspecific pollination, in combination with a relaxed endosperm balance requirement, is the most likely route to the persistence of these populations. We propose that Avon Gorge Sorbus represents a model system for studying the establishment and persistence of SI apomicts in natural populations. PMID

  6. Tiny bubbles challenge giant turbines: Three Gorges puzzle.

    PubMed

    Li, Shengcai

    2015-10-06

    Since the birth of the first prototype of the modern reaction turbine, cavitation as conjectured by Euler in 1754 always presents as a challenge. Following his theory, the evolution of modern reaction (Francis and Kaplan) turbines has been completed by adding the final piece of the element 'draft-tube' that enables turbines to explore water energy at efficiencies of almost 100%. However, during the last two and a half centuries, with increasing unit capacity and specific speed, the problem of cavitation has been manifested and complicated by the draft-tube surges rather than being solved. Particularly, during the last 20 years, the fierce competition in the international market for extremely large turbines with compact design has encouraged the development of giant Francis turbines of 700-1000 MW. The first group (24 units) of such giant turbines of 700 MW each was installed in the Three Gorges project. Immediately after commission, a strange erosion phenomenon appeared on the guide vane of the machines that has puzzled professionals. From a multi-disciplinary analysis, this Three Gorges puzzle could reflect an unknown type of cavitation inception presumably triggered by turbulence production from the boundary-layer streak transitional process. It thus presents a fresh challenge not only to this old turbine industry, but also to the fundamental sciences.

  7. Geomorphological evolution of volcanic fluvial channels: Eighteen years of morphological monitoring of the upper strect of the Tenenepanco Gorge, Popocatépetl volcano, Mexico

    NASA Astrophysics Data System (ADS)

    Tanarro, Luis Miguel; Juan Zamorano, Jose; Andres, Nuria; Palacios, David

    2015-04-01

    During volcanic eruptions a significant volume of material accumulates on the slopes and pre-existing gorges of the stratovolcanoes. This abundance of loose and unconsolidated material is very likely to be mobilized by rapid flows or lahars generated by sudden heavy rain or melting snow and ice. Thus, volcanic gorges are affected by complex cycles of incision, filling and widening, altering the equilibrium of river systems due to the major changes that lahars cause in channel morphology. These geomorphological dynamics characterize the gorges located on the north flank of the Popocatépetl volcano (19°02' N, 98°62' W, 5424 m). This volcano, located in the centre of the Trans-Mexican Volcanic Belt, began its most recent eruptive period in December 1994, when a glacier partially covered the northern slope. Since then, the interaction of volcanic and glacier activity triggered the formation of lahars in the gorges, causing significant morphological changes in the channel (especially in April 1995, July 1997 and January 2001). The most recent major eruption at Popocatépetl took place on 19 July 2003, and since then a series of smaller eruptions has reduced the glacier to near extinction. The aim of this study is to assess the morphological response of the Tenenepanco channel over an 18-year period, from 1995-2013, where two main scenarios can be observed: a) the period from 1995 to 2001 of volcanic activity and glacier retreat with the formation of flows and b) the period from 2002 to 2013 of relative volcanic calm, the almost complete extinction of the glacier, and the formation of secondary lahars associated with heavy rainfall. Monitoring of the gorge has consisted in the elaboration of 14 geomorphological maps during field studies (November 14, 1995, December 5, 1997, February 7, 1998, October 6, 2001, November 14, 1995, December 5, 1997, February 7, 1998, October 6, 2001, Julio 16, 2002, February 11, 2004, September 8, 2004, February 5, 2006, November 2, 2008

  8. Ecology. Three-Gorges Dam--experiment in habitat fragmentation?

    PubMed

    Wu, Jianguo; Huang, Jianhui; Han, Xingguo; Xie, Zongqiang; Gao, Xianming

    2003-05-23

    Habitat fragmentation is the primary cause of the loss of biodiversity and ecosystem services, but its underlying processes and mechanisms remain poorly understood. Studies of islands and insular terrestrial habitats are essential for improving our understanding of habitat fragmentation. We argue that the Three-Gorges Dam, the largest that humans have ever created, presents a unique grand-scale natural experiment that allows ecologists to address a range of critical questions concerning the theory and practice of biodiversity conservation.

  9. Urban impacts on the water quality of selected water bodies in the Three Gorges Reservoir, China

    NASA Astrophysics Data System (ADS)

    Reid, Lucas; Holbach, Andreas; Wei, Hu; Wang, Lijing; Chen, Hao; Zheng, Binghui; Norra, Stefan

    2013-04-01

    Urban systems belong to the major input sources for pollutants into aquatic systems. In China, the rising urbanisation and industrialisation causes a growing pressure on rivers, lakes and estuaries. With the recent impoundment of the Yangtze River by the Three Gorges Dam, the newly formed Three Gorges Reservoir is additionally experiencing drastic changes in the flow regime [1]. In the frame of the Sino-German "Yangtze-Project" [2] samples were taken from the water bodies in proximity to the Cities of Chongqing, Kaixian and Wushan during a field campaign in April 2011. Water samples were analysed for inorganic contents in suspended solids and the dissolved phase to assess the impact of these cities on the water quality of the reservoir. Results show that input from urban sources, together with the effects from the impoundment of the Yangtze River, deteriorates the quality of water and sediments in the Three Gorges Reservoir. Water in the Wushan Lake is trapped in by the Yangtze River flowing by, which leads to longer retention times of effluent water from the city. The chemical composition of the lake water is also measurable upstream in the Daninghe itself and might be due to the backwater effect. In the Xiaojiang River near Kaixian the low flow velocity from the backwater effect of the Yangtze, together with influences from the city have led to problems with algal blooms. High metal concentrations at Chongqing indicate a strong impact of this megacity on the water quality of the Three Gorges Reservoir and the sediments of the Yangtze River. Acknowledgements: Financial support by the Federal Ministry of Education and Research of Germany (BMBF), the Ministry of Science and Technology of the People's Republic of China (MOST) and the German Academic Exchange Service (DAAD). References: [1] Ministry of Environmental Protection of the People's Republic of China, 2010: Bulletin on the Ecological and Environmental Monitoring Results of the Three Gorges Project 2010 [2

  10. Tiny bubbles challenge giant turbines: Three Gorges puzzle

    PubMed Central

    Li, Shengcai

    2015-01-01

    Since the birth of the first prototype of the modern reaction turbine, cavitation as conjectured by Euler in 1754 always presents as a challenge. Following his theory, the evolution of modern reaction (Francis and Kaplan) turbines has been completed by adding the final piece of the element ‘draft-tube’ that enables turbines to explore water energy at efficiencies of almost 100%. However, during the last two and a half centuries, with increasing unit capacity and specific speed, the problem of cavitation has been manifested and complicated by the draft-tube surges rather than being solved. Particularly, during the last 20 years, the fierce competition in the international market for extremely large turbines with compact design has encouraged the development of giant Francis turbines of 700–1000 MW. The first group (24 units) of such giant turbines of 700 MW each was installed in the Three Gorges project. Immediately after commission, a strange erosion phenomenon appeared on the guide vane of the machines that has puzzled professionals. From a multi-disciplinary analysis, this Three Gorges puzzle could reflect an unknown type of cavitation inception presumably triggered by turbulence production from the boundary-layer streak transitional process. It thus presents a fresh challenge not only to this old turbine industry, but also to the fundamental sciences. PMID:26442144

  11. 3D hydrogeological model of the Lower Yarmouk Gorge, Jordan Rift Valley

    NASA Astrophysics Data System (ADS)

    Magri, Fabien; Inbar, Nimrod; Möller, Peter; Raggad, Marwan; Rödiger, Tino; Rosenthal, Eliahu; Shentsis, Izabela; Tzoufka, Kalliopi; Siebert, Christian

    2017-04-01

    The Lower Yarmouk Gorge (LYG) lies on the eastern margin of the lower Jordan Rift Valley (JRV), bounded to the south by the Ajlun and to the north by the Golan Heights. It allows the outflow of the Yarmouk drainage basin and flow into the Jordan River, a few kilometers south of Lake Tiberias. The main aquifer system of the LYG is built mostly of Cretaceous sandstones and carbonates confined by Maastrichtian aquiclude. Fissures allow hydraulic connections between the major water-bearing formations from Quaternary to Upper Cretaceous age. It is supposed that the gorge acts as the mixing zone of two crossing flow pathways: N-S from the Hermon Mountains and from the Ajlun Dome, and E-W from Jebel al Arab Mountain in Syria (also known as Huran Plateau or Yarmouk drainage basin). As a result, several springs can be found within the gorge. These are characterized by widespread temperatures (20 - 60 °C) which indicate that, beside the complex regional flow, also ascending thermal waters control the hydrologic behavior of the LYG. Previous simulations based on a conceptual simplified 3D model (Magri et al., 2016) showed that crossing flow paths result from the coexistence of convection, that can develop for example along NE-SW oriented faults within the gorge or in permeable aquifers below Maastrichtian aquiclude, and additional flow fields that are induced by the N-S topographic gradients. Here we present the first 3D hydrogeological model of the entire LYG that includes structural features based on actual logs and interpreted seismic lines from both Israeli and Jordanian territories. The model distinguishes seven units from upper Eocene to the Lower Triassic, accounting for major aquifers, aquicludes and deep-cutting faults. Recharges are implemented based on the numerical representation developed by Shentsis (1990) that considers relationships between mean annual rain and topographic elevation. The model reveals that topography-driven N-S and E-W flows strongly control

  12. Occurrence of organotin compounds in river sediments under the dynamic water level conditions in the Three Gorges Reservoir Area, China.

    PubMed

    Gao, Jun-Min; Zhang, Ke; Chen, You-Peng; Guo, Jin-Song; Wei, Yun-Mei; Jiang, Wen-Chao; Zhou, Bin; Qiu, Hui

    2015-06-01

    The Three Gorges Project is the largest hydro project in the world, and the water level of the Three Gorges Reservoir (TGR) is dynamic and adjustable with the aim of flood control and electrical power generation. It is necessary to investigate the pollutants and their underlying contamination processes under dynamic water levels to determine their environmental behaviors in the Three Gorges Reservoir Area (TGRA). Here, we report the assessment of organotin compounds (OTs) pollution in the river sediments of the TGRA. Surface sediment samples were collected in the TGRA at low and high water levels. Tributyltin (TBT), triphenyltin (TPhT), and their degradation products in sediments were quantified by gas chromatography-mass spectrometry. Butyltins (BTs) and phenyltins (PhTs) were detected in sediments, and BTs predominated over PhTs in the whole study area under dynamic water level conditions. The concentrations of OTs in sediments varied markedly among locations, and significant concentrations were found in river areas with high levels of boat traffic and wastewater discharge. Sediments at all stations except Cuntan were lightly contaminated with TBT, and total organic carbon (TOC) was a significant factor affecting the fate of TBT in the TGRA. The butyltin and phenyltin degradation indices showed no recent inputs of TBT or TPhT into this region, with the exception of fresh TPhT input at Xiakou Town. Shipping activity, wastewater discharge, and agriculture are the most likely sources of OTs in the TGRA.

  13. Mechanism of Acetylcholinesterase Inhibition by Fasciculin: A 5-ns Molecular Dynamics Simulation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tai, Kaihsu; Shen, T Y.; Henchman, Richard H.

    Our previous molecular dynamics simulation (10 ns) of mouse acetylcholinesterase (EC 3.1.1.7) revealed complex fluctuation of the enzyme active site gorge. Now we report a 5-ns simulation of acetylcholinesterase complexed with fasciculin 2. Fasciculin 2 binds to the gorge entrance of acetylcholinesterase with excellent complementarity and many polar and hydrophobic interactions. In this simulation of the protein-protein complex, where fasciculin 2 appears to sterically block access of ligands to the gorge, again we observe a two-peaked probability distribution of the gorge width. When fasciculin is present, the gorge width distribution is altered such that the gorge is more likely tomore » be narrow. Moreover, there are large increases in the opening of alternative passages, namely, the side door (near Thr 75) and the back door (near Tyr 449). Finally, the catalytic triad arrangement in the acetylcholinesterase active site is disrupted with fasciculin bound. These data support that, in addition to the steric obstruction seen in the crystal structure, fasciculin may inhibit acetylcholinesterase by combined allosteric and dynamical means. Additional data from these simulations can be found at http://mccammon.ucsd.edu/.« less

  14. Recovery of the Three-Gorges Reservoir Impoundment Signal from ICESat altimetry and GRACE

    NASA Astrophysics Data System (ADS)

    Carabajal, C. C.; Boy, J.; Luthcke, S. B.; Harding, D. J.; Rowlands, D. D.; Lemoine, F. G.

    2006-12-01

    The Three Gorges Dam along the Yangtze River in China is one of the largest dams in the world. The water impoundment of the Three-Gorges Reservoir started in June 2003, and the volume of water will continuously increase up to about 40 km3 in 2009, over a length of about 600 km. Although water-level changes along the Yangtze River and the Three Gorges Reservoir are measured by in situ water gauges, access to these data can be quite difficult. Estimates of inland water height and extent can also be recovered from altimetry measurements performed from satellite platforms, such as those acquired by the Geoscience laser Altimetry System (GLAS) on board the Ice, Cloud and Land Elevation Satellite (ICESat). ICESat has produced a comprehensive, highly precise, set of along-track elevation measurements, every three months since its launch in 2003, which intersect the Yangtze River along its East-West extent. In addition, the water impoundment of major artificial reservoirs induces variations of global geodetic quantities, such as the gravity field and Earth rotation (Chao, 1995, Boy & Chao, 2002). Water level changes within the reservoir are compared to GRACE (Gravity Recovery And Climate Experiment) recovered water mass changes. In addition, we compare the GRACE observations of mass change in the Yangtze region to hydrological changes computed from different global soil-moisture and snow models, such as GLDAS (Global Land Data Assimilation System).

  15. Susceptibility of the Batoka Gorge hydroelectric scheme to climate change

    NASA Astrophysics Data System (ADS)

    Harrison, Gareth P.; Whittington, H.(Bert) W.

    2002-07-01

    The continuing and increased use of renewable energy sources, including hydropower, is a key strategy to limit the extent of future climate change. Paradoxically, climate change itself may alter the availability of this natural resource, adversely affecting the financial viability of both existing and potential schemes. Here, a model is described that enables the assessment of the relationship between changes in climate and the viability, technical and financial, of hydro development. The planned Batoka Gorge scheme on the Zambezi River is used as a case study to validate the model and to predict the impact of climate change on river flows, electricity production and scheme financial performance. The model was found to perform well, given the inherent difficulties in the task, although there is concern regarding the ability of the hydrological model to reproduce the historic flow conditions of the upper Zambezi Basin. Simulations with climate change scenarios illustrate the sensitivity of the Batoka Gorge scheme to changes in climate. They suggest significant reductions in river flows, declining power production, reductions in electricity sales revenue and consequently an adverse impact on a range of investment measures.

  16. [Study on species and distribution of flora of national rare and endangered medicinal plant in the Three Gorges area].

    PubMed

    Chen, Shao-Cheng

    2013-04-01

    According to the China Plant Red Data Book and National Key Protected Wild Plants, the distribution of the rare and endangered plants and national conservative plants in the Three Gorges area were investigated and statistically analyzed. Its floristic composition and characteristics of geographical distribution were explored. As a result, a total of 97 species of medicinal flora belonging to rare and endangered national protection plants were found in the Three Gorges area. They come from 81 genera of 46 families. Their vertical distribution is obvious and horizontal distribution has discontinuous overlap. There are many ancient relict medicinal plants in the Three Gorges area. These medicinal plants have obvious temperate characteristics, and are easily found at warm and moist ravines and hillsides; The proportion of tree is much higher than that of herb, vine, shrub and fern. Most of them belong to specific and monotypic genera.

  17. The Dry Season Shuffle: Gorges Provide Refugia for Animal Communities in Tropical Savannah Ecosystems

    PubMed Central

    Doody, J. Sean; Clulow, Simon; Kay, Geoff; D’Amore, Domenic; Rhind, David; Wilson, Steve; Ellis, Ryan; Castellano, Christina; McHenry, Colin; Quayle, Michelle; Hands, Kim; Sawyer, Graeme; Bass, Michael

    2015-01-01

    In the wet-dry tropics, animal species face the major challenges of acquiring food, water or shelter during an extended dry season. Although large and conspicuous animals such as ungulates and waterfowl migrate to wetter areas during this time, little is known of how smaller and more cryptic animal species with less mobility meet these challenges. We fenced off the entire entrance of a gorge in the Australian tropical savanna, offering the unique opportunity to determine the composition and seasonal movement patterns of the small vertebrate community. The 1.7 km-long fence was converted to a trapline that was deployed for 18-21 days during the early dry season in each of two years, and paired traps on both sides of the fence allowed us to detect the direction of animal movements. We predicted that semi-aquatic species (e.g., frogs and turtles) would move upstream into the wetter gorge during the dry season, while more terrestrial species (e.g., lizards, snakes, mammals) would not. The trapline captured 1590 individual vertebrates comprising 60 species. There was a significant bias for captures on the outside of the fence compared to the inside for all species combined (outside/inside = 5.2, CI = 3.7-7.2), for all vertebrate classes, and for specific taxonomic groups. The opposite bias (inside/outside = 7.3, N= 25) for turtles during the early wet season suggested return migration heading into the wet season. Our study revealed that the small vertebrate community uses the gorge as a dry season refuge. The generality of this unreplicated finding could be tested by extending this type of survey to tropical savannahs worldwide. A better understanding of how small animals use the landscape is needed to reveal the size of buffer zones around wetlands required to protect both semi-aquatic and terrestrial fauna in gorges in tropical savannah woodland, and thus in ecosystems in general. PMID:26135472

  18. Competitive Inhibition Mechanism of Acetylcholinesterase without Catalytic Active Site Interaction: Study on Functionalized C60 Nanoparticles via in Vitro and in Silico Assays.

    PubMed

    Liu, Yanyan; Yan, Bing; Winkler, David A; Fu, Jianjie; Zhang, Aiqian

    2017-06-07

    Acetylcholinesterase (AChE) activity regulation by chemical agents or, potentially, nanomaterials is important for both toxicology and pharmacology. Competitive inhibition via direct catalytic active sites (CAS) binding or noncompetitive inhibition through interference with substrate and product entering and exiting has been recognized previously as an AChE-inhibition mechanism for bespoke nanomaterials. The competitive inhibition by peripheral anionic site (PAS) interaction without CAS binding remains unexplored. Here, we proposed and verified the occurrence of a presumed competitive inhibition of AChE without CAS binding for hydrophobically functionalized C 60 nanoparticles (NPs) by employing both experimental and computational methods. The kinetic inhibition analysis distinguished six competitive inhibitors, probably targeting the PAS, from the pristine and hydrophilically modified C 60 NPs. A simple quantitative nanostructure-activity relationship (QNAR) model relating the pocket accessible length of substituent to inhibition capacity was then established to reveal how the geometry of the surface group decides the NP difference in AChE inhibition. Molecular docking identified the PAS as the potential binding site interacting with the NPs via a T-shaped plug-in mode. Specifically, the fullerene core covered the enzyme gorge as a lid through π-π stacking with Tyr72 and Trp286 in the PAS, while the hydrophobic ligands on the fullerene surface inserted into the AChE active site to provide further stability for the complexes. The modeling predicted that inhibition would be severely compromised by Tyr72 and Trp286 deletions, and the subsequent site-directed mutagenesis experiments proved this prediction. Our results demonstrate AChE competitive inhibition of NPs without CAS participation to gain further understanding of both the neurotoxicity and the curative effect of NPs.

  19. Molecular Dynamics of Mouse Acetylcholinesterase Complexed with Huperzine A

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tara, Sylvia; Helms, Volkhard H.; Straatsma, TP

    1999-03-16

    Two molecular dynamics simulations were performed for a modeled complex of mouse acetylcholinesterase liganded with huperzine A (HupA). Analysis of these simulations shows that HupA shifts in the active site toward Tyr 337 and Phe 338, and that several residues in the active site area reach out to make hydrogen bonds with the inhibitor. Rapid fluctuations of the gorge width are observed, ranging from widths that allow substrate access to the active site, to pinched structures that do not allow access of molecules as small as water. Additional openings or channels to the active site are found. One opening ismore » formed in the side wall of the active site gorge by residues Val 73, Asp 74, Thr 83, Glu 84, and Asn 87. Another opening is formed at the base of the gorge by residues Trp 86, Val 132, Glu 202, Gly 448, and Ile 451. Both of these openings have been observed separately in the Torpedo californica form of the enzyme. These channels could allow transport of waters and ions to and from the bulk solution.« less

  20. The Three Gorges Dam of China: Technology to Bridge Two Centuries

    ERIC Educational Resources Information Center

    Wahby, Wafeek S.

    2003-01-01

    Some of the most sophisticated 20th-century technologies have been applied to build the largest hydroelectric dam in the world, the Three Gorges Dam Project (TGDP) of China. The author administered a study abroad course in China from May 27 to June 10, 2000, to study the massive project as it approached the halfway mark of its second and most…

  1. Impact of the Three Gorges Dam on the spawning stock and natural reproduction of Chinese sturgeon in Changjiang River, China

    NASA Astrophysics Data System (ADS)

    Gao, Xin; Lin, Pengcheng; Li, Mingzheng; Duan, Zhonghua; Liu, Huanzhang

    2016-09-01

    Chinese sturgeon ( Acipenser sinensis) is the flagship species of the Changjiang River. The migration route of this species is blocked by the first dam, the Gezhou Dam, and its reproduction is affected by the Three Gorges Dam (TGD), one of the largest dams in the world. We studied the impact of the impoundment of the Three Gorges Reservoir (TGR) since 2003 on the spawning stock and the natural reproduction of the Chinese sturgeon by using our monitoring data from 1997 to 2013. Results indicate that TGR impoundment has delayed the first spawning dates of the fish from middle-late October to late November, decreased the amount of spawning activities from twice to only once each year, and significantly reduced egg production. In particular, the fish did not demonstrate any spawning activities in 2013. Therefore, TGR impoundment significantly affects the natural reproduction of the fish downstream of the TGD. The spawning stock size of the fish is also predicted to further decrease in the future, which will lead to a risk of population extinction. Ecological regulations must be imposed on decreasing the water temperature to 20°C before mid-October and increasing water discharge downstream of the TGD in October to induce spawning of the Chinese sturgeon.

  2. [Research progress on remote sensing of ecological and environmental changes in the Three Gorges Reservoir area, China].

    PubMed

    Teng, Ming-jun; Zeng, Li-xiong; Xiao, Wen-fa; Zhou, Zhi-xiang; Huang, Zhi-lin; Wang, Peng-cheng; Dian, Yuan-yong

    2014-12-01

    The Three Gorges Reservoir area (TGR area) , one of the most sensitive ecological zones in China, has dramatically changes in ecosystem configurations and services driven by the Three Gorges Engineering Project and its related human activities. Thus, understanding the dynamics of ecosystem configurations, ecological processes and ecosystem services is an attractive and critical issue to promote regional ecological security of the TGR area. The remote sensing of environment is a promising approach to the target and is thus increasingly applied to and ecosystem dynamics of the TGR area on mid- and macro-scales. However, current researches often showed controversial results in ecological and environmental changes in the TGR area due to the differences in remote sensing data, scale, and land-use/cover classification. Due to the complexity of ecological configurations and human activities, challenges still exist in the remote-sensing based research of ecological and environmental changes in the TGR area. The purpose of this review was to summarize the research advances in remote sensing of ecological and environmental changes in the TGR area. The status, challenges and trends of ecological and environmental remote-sensing in the TGR area were further discussed and concluded in the aspect of land-use/land-cover, vegetation dynamics, soil and water security, ecosystem services, ecosystem health and its management. The further researches on the remote sensing of ecological and environmental changes were proposed to improve the ecosystem management of the TGR area.

  3. Geological constraints on cave development in the plateau-gorge karst of South China (Wulong, Chongqing)

    NASA Astrophysics Data System (ADS)

    Szczygieł, Jacek; Golicz, Mateusz; Hercman, Helena; Lynch, Erin

    2018-03-01

    The Houping Tiankeng cluster is a part of the South China Karst UNESCO World Natural Heritage Site. Within the distinctive Wulong plateau-gorge karst, > 200 km of cave passages have been documented to date. This paper focuses on detailed tectonic and morphological research on the Luo Shui Kong cave, enriched with U-series dating of speleothems and complemented by morphometric analysis of the San Wang Dong and Er Wang Dong caves. All of these caves exhibit three regional levels of cave development: 1) 1040-1020 m a.s.l.; 2) 900-840 m a.s.l.; and 3) 740-660 m a.s.l. The Houping Tiankeng area is a carbonate rock sequence several hundred meters thick, overlain by the less soluble Lower Ordovician strata, limiting recharge points to faults exposing underlying easily soluble formations. This leads to the domination of concentrated, high-volume inflow and thus results in caves of large volume in the plateau-gorge karst. Shafts connecting the surface with cave passages located underneath formed along faults, changing the hydrogeological pattern through karst water capture and remodeling of existing conduits, albeit mainly by increasing their overall dimensions rather than by deepening them. The most favorable structures for cave-level development are two sets of joints conjugated with gently inclined bedding. Since these joints are characterized by a small vertical extent, downward development is limited. Hence, most of the passages are wide but not deep canyons and typical of a water-table cave pattern. Places where the fault plane is eroded from the surface and where, at the same time, an underneath cave chamber ceiling expands upwards are particularly predisposed to the formation of a tiankeng.

  4. The Evolving Landscape of the Columbia River Gorge: Lewis and Clark and Cataclysms on the Columbia

    USGS Publications Warehouse

    O'Connor, James E.

    2004-01-01

    TAVELERS RETRACING LEWIS AND CLARKE JOURNEY to the Pacific over the past two hundred years have witnessed tre mendous change to the Columbia River Gorge and its pri mary feature, the Columbia River. Dams, reservoirs, timber harvest, altered fisheries, transportation infrastructure, and growth and shrinkage of communities have transformed the river and valley.1 This radically different geography of human use and habitation is commonly contrasted with the sometimes romantic view of a prior time provided both by early nineteenth-century chroniclers and present-day critics of the modern condition ? an ecotopia of plentiful and perpetual resources sustaining a stable culture from time immemorial. Reality is more com plicated. Certainly the human-caused changes to the Columbia River and the gorge since Lewis and Clark have been profound; but the geologic his tory of immense floods, landslides, and volcanic eruptions that occurred before their journey had equally, if not more, acute effects on landscapes and societies of the gorge. In many ways, the Lewis and Clark Expedi tion can be viewed as a hinge point for the Columbia River, the changes engineered to the river and its valley in the two hundred years since their visit mirrored by tremendous changes geologically engendered in the thousands of years before. 

  5. The Evolving Landscape of the Columbia River Gorge: Lewis and Clark and Cataclysms on the Columbia

    USGS Publications Warehouse

    O'Connor, James E.

    2004-01-01

    Travelers reacting Lewis and Clark's journey to the Pacific over the past two hundred years have witnessed tremendous change to the Columbia River Gorge and its primary feature, the Columbia River. Dams, reservoirs, timer harvest, altered fisheries, transportation infrastructure, and growth and shrinkage of communities have transformed the river and valley. This radically different geography of human use and habitation is  commonly contrasted with the sometimes romantic view of a prior time provided both by early nineteenth-century chronicle and present day critics of the modern condition - an ectopia of plentiful and perpetual resources sustaining a stable culture from time immemorial. Reality is more complicated. Certainly the human-caused changes to the Columbia River and the gorge since Lewis and Clark have been profound; by the geologic history of immense floods, landslides, and volcanic eruptions that occurred before the journey had equally, if not more, acute effects on landscapes and societies of the gorge. In many ways, the Lewis and Clark Expidition can be viewed as a hinge point for the Columbia River, the changes engineered to the river and its valley in the two hundred years since their visit mirrored by tremendous cchanges geologically engendered in the thousands of years before. 

  6. [Temporal-spatial analysis of bacillary dysentery in the Three Gorges Area of China, 2005-2016].

    PubMed

    Zhang, P; Zhang, J; Chang, Z R; Li, Z J

    2018-01-10

    Objective: To analyze the spatial and temporal distributions of bacillary dysentery in Chongqing, Yichang and Enshi (the Three Gorges Area) from 2005 to 2016, and provide evidence for the disease prevention and control. Methods: The incidence data of bacillary dysentery in the Three Gorges Area during this period were collected from National Notifiable Infectious Disease Reporting System. The spatial-temporal scan statistic was conducted with software SaTScan 9.4 and bacillary dysentery clusters were visualized with software ArcGIS 10.3. Results: A total of 126 196 cases were reported in the Three Gorges Area during 2005-2016, with an average incidence rate of 29.67/100 000. The overall incidence was in a downward trend, with an average annual decline rate of 4.74%. Cases occurred all the year round but with an obvious seasonal increase between May and October. Among the reported cases, 44.71% (56 421/126 196) were children under 5-year-old, the cases in children outside child care settings accounted for 41.93% (52 918/126 196) of the total. The incidence rates in districts of Yuzhong, Dadukou, Jiangbei, Shapingba, Jiulongpo, Nanan, Yubei, Chengkou of Chongqing and districts of Xiling and Wujiagang of Yichang city of Hubei province were high, ranging from 60.20/100 000 to 114.81/100 000. Spatial-temporal scan statistic for the spatial and temporal distributions of bacillary dysentery during this period revealed that the temporal distribution was during May-October, and there were 12 class Ⅰ clusters, 35 class Ⅱ clusters, and 9 clusters without statistical significance in counties with high incidence. All the class Ⅰ clusters were in urban area of Chongqing (Yuzhong, Dadukou, Jiangbei, Shapingba, Jiulongpo, Nanan, Beibei, Yubei, Banan) and surrounding counties, and the class Ⅱ clusters transformed from concentrated distribution to scattered distribution. Conclusions: Temporal and spatial cluster of bacillary dysentery incidence existed in the three gorges

  7. Wet deposition and sources of inorganic nitrogen in the Three Gorges Reservoir Region, China.

    PubMed

    Wang, Huanbo; Shi, Guangming; Tian, Mi; Chen, Yang; Qiao, Baoqing; Zhang, Liuyi; Yang, Fumo; Zhang, Leiming; Luo, Qiong

    2018-02-01

    Precipitation samples were collected at five rural and one urban sites in the Three Gorges Reservoir Region (TGR), China from March 2014 to February 2016. The inorganic reactive nitrogen (Nr) contents were analysed to investigate their wet deposition flux, budget, and sources in the area. Annual Nr wet deposition varied from 7.1 to 23.4 kg N ha -1 yr -1 over the six sites during the two-year study campaign. The six-site average Nr wet deposition flux was 17.1 and 11.7 kg N ha -1 yr -1 in 2014 and 2015, respectively, with 71% from NH 4 + and 29% from NO 3 - . Dry deposition flux was estimated using the inferential method, which combined the measured ambient concentrations and modelled dry deposition velocities. The total (dry + wet) Nr deposition fluxes were estimated to be 21.4 kg N ha -1 yr -1 in 2014 and 16.0 kg N ha -1 yr -1 in 2015 at rural sites, and 31.4 and 25.3 kg N ha -1 yr -1 at the urban site. Annual average volume weighted mean (VWM) concentrations in precipitation at all the six sites differed little for NO 3 - but up to a factor of 2.0 for NH 4 + with the highest value at the urban site. Industrial emissions, agricultural emissions, soil dust, and biomass burning were identified as potential sources of the major inorganic ions in precipitation using factor analysis and correlation analysis. Conditional probability function (CPF) analysis indicated that the urban site was predominantly affected by industrial emissions from a power plant, cement manufactory, and salt chemical facility located ∼13 km southeast of the sampling site. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  8. 3D Coupled Thermal-Hydraulic Model of the Lower Yarmouk Gorge, Jordan Rift Valley

    NASA Astrophysics Data System (ADS)

    Walther, M.; Magri, F.; Inbar, N.; Möller, P.; Raggad, M.; Rödiger, T.; Rosenthal, E.; Shentsis, I.; Siebert, C.; Volpi, G.

    2017-12-01

    It is supposed that the Lower Yarmouk Gorge (LYG), in the Jordan Rift Valley acts as the mixing zone of two crossing flow pathways: N-S from the Hermon Mountains and from the Ajlun Dome, and E-W from Jebel al Arab Mountain in Syria (also known as Huran Plateau or Yarmouk drainage basin). As a result, several springs can be found within the gorge. These are characterized by widespread temperatures (20 - 60 °C) which indicate that, beside the complex regional flow, also ascending thermal waters control the hydrologic behavior of the LYG. Previous simulations based on a conceptual simplified 3D model (Magri et al., 2016) showed that crossing flow paths result from the coexistence of convection, that can develop for example along NE-SW oriented faults within the gorge or in permeable aquifers below Maastrichtian aquiclude, and additional flow fields that are induced by the N-S topographic gradients. Here we present the first 3D hydrogeological model of the entire LYG that includes structural features based on actual logs and interpreted seismic lines from both Israeli and Jordanian territories. The model distinguishes seven units from upper Eocene to the Lower Triassic, accounting for major aquifers, aquicludes and deep-cutting faults. Recharges are implemented based on the numerical representation developed by Shentsis (1990) that considers relationships between mean annual rain and topographic elevation. The model reveals that topography-driven N-S and E-W flows strongly control the location of discharge areas while the anomalous spring temperature is not necessarily linked to the presence of fault convection. Local permeability anisotropy due to aquifers folding or facies changes are features sufficient for the rising of hot fluids. Shentsis, I., 1990. Mathematical models for long-term prediction of mountainous river runoff: methods, information and results, Hydrological Sciences Journal, 35:5, 487-500 Magri, F., Möller, S., Inbar, N., Möller, P., Raggad, M., R

  9. Game-Theory Based Research on Oil-Spill Prevention and Control Modes in Three Gorges Reservoir Area

    NASA Astrophysics Data System (ADS)

    Yin, Jie; Xiong, Ting

    2018-01-01

    Aiming at solving the existing oil pollution in the Three Gorges reservoir, this paper makes research on oil-spill prevention and control mode based on game theory. Regarding the built modes and comparative indicator system, overall efficiency indicator functions are used to compare general effect, overall cost, and overall efficiency, which concludes that the mode combining government and enterprise has the highest overall efficiency in preventing and controlling ship oil spills. The suggested mode together its correspondingly designed management system, has been applied to practice for a year in Three Gorges Reservoir Area and has made evident improvements to the existing oil pollution, meanwhile proved to be quite helpful to the pollution prevention and control in the lower reaches of Yangtze River.

  10. Mouse Acetylcholinesterase Unliganded and in Complex with Huperzine A: A Comparison of Molecular Dynamics Simulations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tara, Sylvia; Straatsma, TP; Mccammon, Andy

    1999-06-01

    A 1 ns molecular dynamics simulation of unliganded mouse acetylcholinesterase (AChE) is compared to a previous simulation of mouse AChE complexed with Huperzine A (HupA). Several common features are observed. In both simulations, the active site gorge fluctuates in size during the 1 ns trajectory, and is completely pinched off several times. Many of the residues in the gorge that formed hydrogen bonds with HupA in the simulation of the complex, now form hydrogen bonds with other protein residues and water molecules in the gorge. The opening of a "backdoor" entrance to the active site that was found in themore » simulation of the complex is also observed in the unliganded simulation. Differences between the two simulations include overall lower structural RMS deviations for residues in the gorge in the unliganded simulation, a smaller diameter of the gorge in the absence of HupA, and the disappearance of a side channel that was frequently present in the liganded simulation. The differences between the two simulations can be attributed, in part, to the interaction of AChE with HupA.« less

  11. Molecular Characterization of Monoclonal Antibodies that Inhibit Acetylcholinesterase by Targeting the Peripheral Site and Backdoor Region

    PubMed Central

    Essono, Sosthène; Mondielli, Grégoire; Lamourette, Patricia; Boquet, Didier; Grassi, Jacques; Marchot, Pascale

    2013-01-01

    The inhibition properties and target sites of monoclonal antibodies (mAbs) Elec403, Elec408 and Elec410, generated against Electrophorus electricus acetylcholinesterase (AChE), have been defined previously using biochemical and mutagenesis approaches. Elec403 and Elec410, which bind competitively with each other and with the peptidic toxin inhibitor fasciculin, are directed toward distinctive albeit overlapping epitopes located at the AChE peripheral anionic site, which surrounds the entrance of the active site gorge. Elec408, which is not competitive with the other two mAbs nor fasciculin, targets a second epitope located in the backdoor region, distant from the gorge entrance. To characterize the molecular determinants dictating their binding site specificity, we cloned and sequenced the mAbs; generated antigen-binding fragments (Fab) retaining the parental inhibition properties; and explored their structure-function relationships using complementary x-ray crystallography, homology modeling and flexible docking approaches. Hypermutation of one Elec403 complementarity-determining region suggests occurrence of antigen-driven selection towards recognition of the AChE peripheral site. Comparative analysis of the 1.9Å-resolution structure of Fab408 and of theoretical models of its Fab403 and Fab410 congeners evidences distinctive surface topographies and anisotropic repartitions of charges, consistent with their respective target sites and inhibition properties. Finally, a validated, data-driven docking model of the Fab403-AChE complex suggests a mode of binding at the PAS that fully correlates with the functional data. This comprehensive study documents the molecular peculiarities of Fab403 and Fab410, as the largest peptidic inhibitors directed towards the peripheral site, and those of Fab408, as the first inhibitor directed toward the backdoor region of an AChE and a unique template for the design of new, specific modulators of AChE catalysis. PMID:24146971

  12. [Impact on prevalence of schitosomiasis after runs of Three Gorges Reservoir Project in the section of Anhui province].

    PubMed

    Zhang, Shiqing; Wang, Tianping; He, Jiachang; Li, Huazhong; Tian, Xuegen; Gao, Fenghua

    2015-07-01

    To investigate the changes of water level and the distribution of snails in Anhui province before and after runs of the Three Gorges Reservoir Project, and to determine the relationship between the two factors and schistosomiasis transmission. The hydrologic data of Datong hydrologic station and the data of snail status and schistosomiasis morbidity in Anhui Province were collected. The data from 1991 to 2002 and 2003 to 2012 were considered as before and after the impoundment of the Three Gorges Reservoir Project. Based on the prevalence of schistosomiasis, the cases of people and cattle were speculated, and the average infection rate of people and cattle were calculated. The t-test was used to compare the difference of snail area and the density of living snails before and after the impoundment of the Three Gorges Project. The pearson method was used to analyze the relationship between water level and snail area. The spearman method was used to analyze the relationship between the water level and the distribution of snails. From 1991 to 2012, the range of the highest water level, the lowest water level, the difference between the highest and lowest water level, the mean in the abundant water seasons, the mean in the dry water seasons, and the difference between the abundant water seasons and the dry water seasons was 11.40-16.30, 3.68-5.20, 6.70-12.12, 9.92-14.40, 4.77-7.64 and 4.13-8.93 m, respectively. The snail areas was (28 613 ± 362) hm² and (29 477 ± 918) hm² (t = -3.00, P = 0.007), the density of living snails was 1.51 (1.15-2.43) and 0.43 (0.29-1.10) snails/0.11 m² (H = 4.28, P < 0.001) before and after the impoundment of the Three Gorges Project, respectively. The average infection rate of people and cattle was 1.68% (99 482/5 935 147) and 4.62% (13 923/3 011 33), and the average number of acute schistosomiasis cases was 328, before the impoundment of the Three Gorges Project; 0.60% (39 747/6 649 380), 1.65% (1 291/783 224) and 71 after the

  13. Lions as Bone Accumulators? Paleontological and Ecological Implications of a Modern Bone Assemblage from Olduvai Gorge.

    PubMed

    Arriaza, Mari Carmen; Domínguez-Rodrigo, Manuel; Yravedra, José; Baquedano, Enrique

    2016-01-01

    Analytic models have been developed to reconstruct early hominin behaviour, especially their subsistence patterns, revealed mainly through taphonomic analyses of archaeofaunal assemblages. Taphonomic research is used to discern which agents (carnivores, humans or both) generate the bone assemblages recovered at archaeological sites. Taphonomic frameworks developed during the last decades show that the only large-sized carnivores in African biomes able to create bone assemblages are leopards and hyenas. A carnivore-made bone assemblage located in the short-grassland ecological unit of the Serengeti (within Olduvai Gorge) was studied. Taphonomic analyses of this assemblage including skeletal part representation, bone density, breakage patterns and anatomical distribution of tooth marks, along with an ecological approach to the prey selection made by large carnivores of the Serengeti, were carried out. The results show that this bone assemblage may be the first lion-accumulated assemblage documented, although other carnivores (namely spotted hyenas) may have also intervened through postdepositional ravaging. This first faunal assemblage potentially created by lions constitutes a new framework for neotaphonomic studies. Since lions may accumulate carcasses under exceptional circumstances, such as those documented at the site reported here, this finding may have important consequences for interpretations of early archaeological and paleontological sites, which provide key information about human evolution.

  14. Lions as Bone Accumulators? Paleontological and Ecological Implications of a Modern Bone Assemblage from Olduvai Gorge

    PubMed Central

    Arriaza, Mari Carmen; Domínguez-Rodrigo, Manuel; Yravedra, José; Baquedano, Enrique

    2016-01-01

    Analytic models have been developed to reconstruct early hominin behaviour, especially their subsistence patterns, revealed mainly through taphonomic analyses of archaeofaunal assemblages. Taphonomic research is used to discern which agents (carnivores, humans or both) generate the bone assemblages recovered at archaeological sites. Taphonomic frameworks developed during the last decades show that the only large-sized carnivores in African biomes able to create bone assemblages are leopards and hyenas. A carnivore-made bone assemblage located in the short-grassland ecological unit of the Serengeti (within Olduvai Gorge) was studied. Taphonomic analyses of this assemblage including skeletal part representation, bone density, breakage patterns and anatomical distribution of tooth marks, along with an ecological approach to the prey selection made by large carnivores of the Serengeti, were carried out. The results show that this bone assemblage may be the first lion-accumulated assemblage documented, although other carnivores (namely spotted hyenas) may have also intervened through postdepositional ravaging. This first faunal assemblage potentially created by lions constitutes a new framework for neotaphonomic studies. Since lions may accumulate carcasses under exceptional circumstances, such as those documented at the site reported here, this finding may have important consequences for interpretations of early archaeological and paleontological sites, which provide key information about human evolution. PMID:27144649

  15. 76 FR 62089 - General Management Plan/Abbreviated Final Environmental Impact Statement, New River Gorge...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-06

    ... Impact Statement for the General Management Plan (GMP/EIS) for New River Gorge National River, West..., and interpretation of park resources over the next 20 years. The Abbreviated Final GMP/EIS includes an analysis of agency and public comments received on the Draft GMP/EIS with NPS responses, errata sheets...

  16. Acetylcholinesterase complexed with bivalent ligands related to huperzine a: experimental evidence for species-dependent protein-ligand complementarity.

    PubMed

    Wong, Dawn M; Greenblatt, Harry M; Dvir, Hay; Carlier, Paul R; Han, Yi-Fan; Pang, Yuan-Ping; Silman, Israel; Sussman, Joel L

    2003-01-15

    Acetylcholinesterase (AChE) inhibitors improve the cognitive abilities of Alzheimer patients. (-)-Huperzine A [(-)-HupA], an alkaloid isolated from the club moss, Huperzia serrata, is one such inhibitor, but the search for more potent and selective drugs continues. Recently, alkylene-linked dimers of 5-amino-5,6,7,8-tetrahydroquinolinone (hupyridone, 1a), a fragment of HupA, were shown to serve as more potent inhibitors of AChE than (-)-HupA and monomeric 1a. We soaked two such dimers, (S,S)-(-)-bis(10)-hupyridone [(S,S)-(-)-2a] and (S,S)-(-)-bis(12)-hupyridone [(S,S)-(-)-2b] containing, respectively, 10 and 12 methylenes in the spacer, into trigonal TcAChE crystals, and solved the X-ray structures of the resulting complexes using the difference Fourier technique, both to 2.15 A resolution. The structures revealed one HupA-like 1a unit bound to the "anionic" subsite of the active-site, near the bottom of the active-site gorge, adjacent to Trp84, as seen for the TcAChE/(-)-HupA complex, and the second 1a unit near Trp279 in the "peripheral" anionic site at the top of the gorge, both bivalent molecules thus spanning the active-site gorge. The results confirm that the increased affinity of the dimeric HupA analogues for AChE is conferred by binding to the two "anionic" sites of the enzyme. Inhibition data show that (-)-2a binds to TcAChE approximately 6-7- and > 170-fold more tightly than (-)-2b and (-)-HupA, respectively. In contrast, previous data for rat AChE show that (-)-2b binds approximately 3- and approximately 2-fold more tightly than (-)-2a and (-)-HupA, respectively. Structural comparison of TcAChE with rat AChE, as represented by the closely related mouse AChE structure (1maa.pdb), reveals a narrower gorge for rat AChE, a perpendicular alignment of the Tyr337 ring to the gorge axis, and its conformational rigidity, as a result of hydrogen bonding between its hydroxyl group and that of Tyr341, relative to TcAChE Phe330. These structural differences in the

  17. Eruption History and Geochemical Evolution of Servilleta Basalt Along the Rio Grande Gorge, Colorado and New Mexico

    NASA Astrophysics Data System (ADS)

    Cosca, M. A.; Thompson, R. A.; Turner, K. J.; Morgan, L. E.

    2016-12-01

    Subalkaline basalt to basaltic andesite lava flows formally known as Servilleta Basalt (SB) are the most voluminous rock type forming the Pliocene Taos Plateau volcanic field. Pleistocene incision by the Rio Grande into the bedrock-floored plateau has resulted in spectacular exposures of occasionally thick ( 240 m) accumulations of SB within the Rio Grande gorge. Incremental CO2 laser heating of individual rock fragments, the SB within and along the length of the Rio Grande gorge has been precisely dated by 40Ar/39Ar geochronology to between 5.3 Ma and 3.3 Ma. SB older than 4 Ma is restricted to some lava flows exposed between La Junta point, at the confluence of the Red River and Rio Grande, and the Gorge Bridge crossing northwest of Taos, NM. Vertical sampling through thick SB flow sequences within the gorge yields precise emplacement histories and also reveals small but systematic major and minor element concentration variations (including Si, Rb, Sr, Cu and Zn). 40Ar/39Ar data show that these trends developed over short (0-250 ka) timescales, and probably relate to partial assimilation of crust, possibly at multiple depths. Combined field, geochemical, and 40Ar/39Ar data consequently record short-lived changes in tholeiitic melt compositions in response to regional extension and development of the Rio Grande rift. The age, lateral extent, and thickness of exposed SB partially reflect the paleotopographic surface of the southern San Luis Basin prior to onset of Pliocene Taos Plateau volcanic field magmatism; paleotopographic highs diverted some flows while topographic lows were areas of infilling and accumulation. Heterogeneous basin paleotopography developed during contemporaneous or precursory andesitic to dacitic volcanism, extensional faulting and subsidence of sub-basins within the San Luis Basin, and deposition of prograding alluvial fans that originated in the Sangre de Cristo and Picuris Mountains. SB flowed into the southern San Luis Valley beginning 5

  18. Red tree voles in the Columbia River Gorge and Hood River basin, Oregon

    Treesearch

    Eric D. Forsman; James K. Swingle; Michael A. McDonald; Scott A. Graham; Nicholas R. Hatch

    2009-01-01

    In 2003 to 2008, we conducted surveys to document the eastern and northern range limits of red tree voles (Arborimus longicaudus) in the Columbia River Gorge and Hood River basin, Oregon. Our survey indicated the current range of the vole includes the area from Wahkeena Creek, 20 km east of Troutdale to Seneca Fouts State Park, 6 km west of Hood...

  19. Statistical evaluation of the effects of fall and winter flows on the spring condition of rainbow and brown trout in the green river downstream of Flaming Gorge Dam.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Magnusson, A. K.; LaGory, K. E.; Hayse, J. W.

    2009-01-09

    Flaming Gorge Dam, a hydroelectric facility operated by the Bureau of Reclamation (Reclamation), is located on the Green River in Daggett County, northeastern Utah. In recent years, single peak releases each day or steady flows have been the operational pattern during the winter period. A double-peak pattern (two flow peaks each day) was implemented during the winter of 2006-2007 by Reclamation. Because there is no recent history of double-peaking at Flaming Gorge Dam, the potential effects of double-peaking operations on the body condition of trout in the dam's tailwater are not known. A study plan was developed that identified researchmore » activities to evaluate potential effects from double-peaking operations during winter months. Along with other tasks, the study plan identified the need to conduct a statistical analysis of existing data on trout condition and macroinvertebrate abundance to evaluate potential effects of hydropower operations. This report presents the results of this analysis. We analyzed historical data to (1) describe temporal patterns and relationships among flows, benthic macroinvertebrate abundance, and condition of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) in the tailwaters of Flaming Gorge Dam and (2) to evaluate the degree to which flow characteristics (i.e., flow volumes and flow variability) and benthic macroinvertebrate abundance affect the condition of trout in this area. This information, together with further analyses of size-stratified trout data, may also serve as baseline data to which the effects of potential future double-peaking flows can be compared. The condition (length, weight and/or relative weight) of rainbow trout (Oncorhynchus mykiss) at two sites in the Green River downstream of Flaming Gorge Dam (Tailrace and Little Hole) and weight of brown trout (Salmo trutta) at the Little Hole site has been decreasing since 1990 while the abundance of brown trout has been increasing at the two sites

  20. A Homo habilis maxilla and other newly-discovered hominid fossils from Olduvai Gorge, Tanzania.

    PubMed

    Clarke, R J

    2012-08-01

    In 1995, a 1.8 million year old hominid maxilla with complete dentition (OH 65) was excavated from Bed I in the western part of Olduvai Gorge. The molar crowns are small relative to the long flaring roots, and the root of the canine is very long and straight. The broad maxilla with wide U-shaped palate and the form of the tooth roots closely match those of KNM-ER 1470 which, in its parietal size and morphology, matches the type specimen of Homo habilis, OH 7. Thus, OH 65 and KNM-ER 1470 group with OH 7 as representatives of H. habilis while some other Olduvai specimens, such as OH 13 and OH 24, have more in common in terms of morphology and brain size with Australopithecus africanus. Between 1995 and 2007, the OLAPP team has recovered teeth of eight other hominid individuals from various parts of Olduvai Gorge. These have been identified as belonging to H. habilis, Paranthropus boisei, and Australopithecus cf. africanus. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. The Short-Term Impact of Involuntary Migration in China's Three Gorges: A Prospective Study

    ERIC Educational Resources Information Center

    Hwang, Sean-Shong; Cao, Yue; Xi, Juan

    2011-01-01

    The aim of this study is to measure the short-term impact of involuntary migration resulting from China's Three Gorges Dam project on the 1.3 million persons being displaced. We focus on the social, economic, and mental and physical health impact using three sets of indicators. Using a prospective research design, we gathered information about…

  2. 75 FR 18877 - Notice of Final Supplementary Rules for Public Lands in Colorado: Gunnison Gorge National...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-13

    ... DEPARTMENT OF THE INTERIOR Bureau of Land Management [LLCOS05400-L17110000 PA000 LXSIGGCA0000] Notice of Final Supplementary Rules for Public Lands in Colorado: Gunnison Gorge National Conservation Area (GGNCA) and Adjacent Public Lands Administered by the Bureau of Land Management Uncompahgre Field...

  3. [Impact of people's social behavior on schistosomiasis transmission in the Three Gorges Reservoir area].

    PubMed

    Zhang, Hui-Juan; Zhu, Rong; Wu, Cheng-Guo; Guo, Jia-Gang

    2007-06-01

    To investigate the changes in natural ecology and social environment after the construction of the dam in the Three Gorges Reservoir area, and to understand the social behavior status and education backgrounds of the local residents, and to analyze the potential impact of these factors on schistosomiasis spreading. Data of nature and social economy after the construction of the dam in the Three Gorge area were collected, and a cross-sectional study with questionnaire survey on information including social demographic characteristics, people's production and life style, knowledge about schistosomiasis was conducted among natives, emigrations. After the construction of the dam, as the stream became slower, the water-level fluctuating zone on the bank of the reservoir was formed. The main source of drinking water and daily life activity are mainly depends on the river supplying (accounting about 68.7% and 75.8%, respectively); due to the needs of farming, washing cloths and vegetables, 45.6% of resident had a close contact with river water. People with away-from-home employment among emigrations were increased from 5.7% to 18.7%. The proportion of using hygienic lavatory was 88.6%. The main source of water for drinking and daily life among emigrations was tap water (85.4% and 87.0%). The residents had no awareness about the health hazard for schistosomiasis, with less health information The natural environment of reservoir areas and the life style are risk factors for the spread of schistosomiasis. The improved sanitation condition and the economy construction after the construction of the dam is propitious to keeping away from the introduction of schistosomiasis. However, people with away-from-home employment among emigrations are still at risk of schistosomiasis.

  4. The Columbia River Basalt Group: from the gorge to the sea

    USGS Publications Warehouse

    Wells, Ray E.; Niem, Alan R.; Evarts, Russell C.; Hagstrum, Jonathan T.

    2009-01-01

    Miocene flood basalts of the Columbia River Basalt Group inundated eastern Washington, Oregon, and adjacent Idaho between 17 and 6 Ma. Some of the more voluminous flows followed the ancestral Columbia River across the Cascade arc, Puget-Willamette trough, and the Coast Range to the Pacific Ocean. We have used field mapping, chemistry, and paleomagnetic directions to trace individual flows and flow packages from the Columbia River Gorge westward into the Astoria Basin, where they form pillow palagonite complexes and mega-invasive bodies into older marine sedimentary rocks. Flows of the Grande Ronde, Wanapum, and Saddle Mountains Basalts all made it to the ocean; at least 33 flows are recognized in the western Columbia River Gorge, 50 in the Willamette Valley, 16 in the lower Columbia River Valley, and at least 12 on the Oregon side of the Astoria Basin. In the Astoria Basin, the basalt flows loaded and invaded the wet marine sediments, producing peperite breccias, soft sediment deformation, and complex invasive relations. Mega-invasive sills up to 500 m thick were emplaced into strata as old as Eocene, and invasive dikes up to 90 m thick can be traced continuously for 25 km near the basin margin. Mega-pillow complexes up to a kilometer thick are interpreted as the remains of lava deltas that prograded onto the shelf and a filled submarine canyon southeast of Astoria, possibly providing the hydraulic head for injection of invasive sills and dikes at depth.

  5. Occurrence and potential health risk of Cryptosporidium and Giardia in the Three Gorges Reservoir, China.

    PubMed

    Xiao, Guosheng; Qiu, Zhiqun; Qi, Junsheng; Chen, Ji-an; Liu, Fengdan; Liu, Wenyi; Luo, Jiaohua; Shu, Weiqun

    2013-05-01

    The Three Gorges Reservoir (TGR) is the biggest lake in the world and a major water source in China. There is no information about occurrence and impact of Cryptosporidium and Giardia on the aquatic ecosystem. 61 surface water samples from 23 monitoring sites and 5 treated effluent samples were collected and analyzed. Cryptosporidium oocysts and Giardia cysts were found, respectively, in 86.4% and 65.2% of a total of 66 water samples, with high concentrations in treated effluent. The mean percent recovery was 29.14% for oocysts and 34.86% for cysts. A seasonal pattern was observed, with positive samples for Cryptosporidium more frequent in flood period and positive samples for Giardia more frequent in impounding period. Counts of enterococci, fecal coliforms and total coliforms, and turbidity were significantly associated with Cryptosporidium concentration in backwater (water in a main river which is backed up by the Three Gorges Dam) areas of tributaries but not Giardia. High associations were also found between oocyst and cyst in backwater areas of tributaries and cities. The risks of infection and illness due to water consumption in four different exposure routes were estimated. The results showed that swimming in the TGR has the highest infection risk with 1.39 × 10(-3) per time (95% confidence interval (CI): 0.05-600.3 × 10(-5)) for Cryptosporidium and 2.08 × 10(-4) per time (95% CI: 0.05-878.87 × 10(-6)) for Giardia, while directly drinking unboiled tap water treated with the conventional process has the highest morbidity with 524.98 per 100,000 population per year (95% CI: 10.35-2040.26) for Cryptosporidium and 5.89 per 100,000 population per year (95% CI: 0.08-22.67) for Giardia. This study provides new useful information for drinking water plants, health care workers and managers to improve the safety of tap water and deduce the risk of surface water contamination in China. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Fire history reflects human history in the Pine Creek Gorge of north-central Pennsylvania

    Treesearch

    Patrick H. Brose; Richard P. Guyette; Joseph M. Marschall; Michael C. Stambaugh

    2015-01-01

    Fire history studies are important tools for understanding past fire regimes and the roles humans played in those regimes. Beginning in 2010, we conducted a fire history study in the Pine Creek Gorge area of north-central Pennsylvania to ascertain the number of fires and fire-free intervals, their variability through time, and the role of human influences. We collected...

  7. Prediction of the binding sites of huperzine A in acetylcholinesterase by docking studies

    NASA Astrophysics Data System (ADS)

    Pang, Yuan-Ping; Kozikowski, Alan P.

    1994-12-01

    We have performed docking studies with the SYSDOC program on acetylcholinesterase (AChE) to predict the binding sites in AChE of huperzine A (HA), which is a potent and selective, reversible inhibitor of AChE. The unique aspects of our docking studies include the following: (i) Molecular flexibility of the guest and the host is taken into account, which permits both to change their conformations upon binding. (ii) The binding energy is evaluated by a sum of energies of steric, electrostatic and hydrogen bonding interactions. In the energy calculation no grid approximation is used, and all hydrogen atoms of the system are treated explicitly. (iii) The energy of cation-π interactions between the guest and the host, which is important in the binding of AChE, is included in the calculated binding energy. (iv) Docking is performed in all regions of the host's binding cavity. Based on our docking studies and the pharmacological results reported for HA and its analogs, we predict that HA binds to the bottom of the binding cavity of AChE (the gorge) with its ammonium group interacting with Trp84, Phe330, Glu199 and Asp72 (catalytic site). At the the opening of the gorge with its ammonium group partially interacting with Trp279 (peripheral site). At the catalytic site, three partially overlapping subsites of HA were identified which might provide a dynamic view of binding of HA to the catalytic site.

  8. Geographical Overview of the Three Gorges Dam and Reservoir, China - Geologic Hazards and Environmental Impacts

    USGS Publications Warehouse

    Highland, Lynn M.

    2008-01-01

    The Three Gorges Dam and Reservoir on the Yangtze River, China, has been an ambitious and controversial project. The dam, the largest in the world as of 2008, will provide hydropower, help to manage flood conditions, and increase the navigability of the Yangtze River. However, this massive project has displaced human and animal populations and altered the stability of the banks of the Yangtze, and it may intensify the seismic hazard of the area. It has also hindered archeological investigations in the reservoir and dam area. This report, originally in the form of a Microsoft PowerPoint presentation, gives a short history and overview of the dam construction and subsequent consequences, especially geologic hazards already noted or possible in the future. The report provides photographs, diagrams, and references for the reader's further research - a necessity, because this great undertaking is dynamic, and both its problems and successes continue to evolve. The challenges and consequences of Three Gorges Dam will be closely watched and documented as lessons learned and applied to future projects in China and elsewhere.

  9. The effects of overwinter flowson the spring condition of rainbow and brown trout size classes in the Green River downstream of Flaming Gorge Dam, Utah.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Magnusson, A. K.; LaGory, K. E.; Hayse, J. W.

    2010-06-25

    Flaming Gorge Dam, a hydroelectric facility operated by the Bureau of Reclamation (Reclamation), is located on the Green River in Daggett County, northeastern Utah. Until recently, and since the early 1990s, single daily peak releases or steady flows have been the operational pattern of the dam during the winter period. However, releases from Flaming Gorge Reservoir followed a double-peak pattern (two daily flow peaks) during the winters of 2006-2007 and 2008-2009. Because there is little recent long-term history of double-peaking at Flaming Gorge Dam, the potential effects of double-peaking operations on trout body condition in the dam's tailwater are notmore » known. A study plan was developed that identified research activities to evaluate potential effects from winter double-peaking operations (Hayse et al. 2009). Along with other tasks, the study plan identified the need to conduct a statistical analysis of historical trout condition and macroinvertebrate abundance to evaluate the potential effects of hydropower operations. The results from analyses based on the combined size classes of trout (85-630 mm) were presented in Magnusson et al. (2008). The results of this earlier analysis suggested possible relationships between trout condition and flow, but concern that some of the relationships resulted from size-based effects (e.g., apparent changes in condition may have been related to concomitant changes in size distribution, because small trout may have responded differently to flow than large trout) prompted additional analysis of within-size class relationships. This report presents the results of analyses of three different size classes of trout (small: 200-299 mm, medium: 300-399 mm, and large: {ge}400 mm body length). We analyzed historical data to (1) describe temporal patterns and relationships among flows, benthic macroinvertebrate abundance, and condition of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) in the tailwaters of

  10. Evaluation of Land Use, Land Management and Soil Conservation Strategies to Reduce Non-Point Source Pollution Loads in the Three Gorges Region, China

    NASA Astrophysics Data System (ADS)

    Strehmel, Alexander; Schmalz, Britta; Fohrer, Nicola

    2016-11-01

    The construction of the Three Gorges Dam in China and the subsequent impoundment of the Yangtze River have induced a major land use change in the Three Gorges Reservoir Region, which fosters increased inputs of sediment and nutrients from diffuse sources into the water bodies. Several government programs have been implemented to mitigate high sediment and nutrient loads to the reservoir. However, institutional weaknesses and a focus on economic development have so far widely counteracted the effectiveness of these programs. In this study, the eco-hydrological model soil and water assessment tool is used to assess the effects of changes in fertilizer amounts and the conditions of bench terraces in the Xiangxi catchment in the Three Gorges Reservoir Region on diffuse matter releases. With this, the study aims at identifying efficient management measures, which should have priority. The results show that a reduction of fertilizer amounts cannot reduce phosphorus loads considerably without inhibiting crop productivity. The condition of terraces in the catchment has a strong impact on soil erosion and phosphorus releases from agricultural areas. Hence, if economically feasible, programmes focusing on the construction and maintenance of terraces in the region should be implemented. Additionally, intercropping on corn fields as well as more efficient fertilization schemes for agricultural land were identified as potential instruments to reduce diffuse matter loads further. While the study was carried out in the Three Gorges Region, its findings may also beneficial for the reduction of water pollution in other mountainous areas with strong agricultural use.

  11. Evaluation of Land Use, Land Management and Soil Conservation Strategies to Reduce Non-Point Source Pollution Loads in the Three Gorges Region, China.

    PubMed

    Strehmel, Alexander; Schmalz, Britta; Fohrer, Nicola

    2016-11-01

    The construction of the Three Gorges Dam in China and the subsequent impoundment of the Yangtze River have induced a major land use change in the Three Gorges Reservoir Region, which fosters increased inputs of sediment and nutrients from diffuse sources into the water bodies. Several government programs have been implemented to mitigate high sediment and nutrient loads to the reservoir. However, institutional weaknesses and a focus on economic development have so far widely counteracted the effectiveness of these programs. In this study, the eco-hydrological model soil and water assessment tool is used to assess the effects of changes in fertilizer amounts and the conditions of bench terraces in the Xiangxi catchment in the Three Gorges Reservoir Region on diffuse matter releases. With this, the study aims at identifying efficient management measures, which should have priority. The results show that a reduction of fertilizer amounts cannot reduce phosphorus loads considerably without inhibiting crop productivity. The condition of terraces in the catchment has a strong impact on soil erosion and phosphorus releases from agricultural areas. Hence, if economically feasible, programmes focusing on the construction and maintenance of terraces in the region should be implemented. Additionally, intercropping on corn fields as well as more efficient fertilization schemes for agricultural land were identified as potential instruments to reduce diffuse matter loads further. While the study was carried out in the Three Gorges Region, its findings may also beneficial for the reduction of water pollution in other mountainous areas with strong agricultural use.

  12. Flood characteristics for the New River in the New River Gorge National River, West Virginia

    USGS Publications Warehouse

    Wiley, J.B.; Cunningham, M.K.

    1994-01-01

    The frequency and magnitude of flooding of the New River in the New River Gorge National River was studied. A steady-state, one-dimensional flow model was applied to the study reach. Rating curves, cross sections, and Manning's roughness coefficients that were used are presented in this report. Manning's roughness coefficients were evaluated by comparing computed elevations (from application of the steady-state, one-dimensional flow model) to rated elevations at U.S. Geological Survey (USGS) streamflow-gaging stations and miscellaneous-rating sites. Manning's roughness coefficients ranged from 0.030 to 0.075 and varied with hydraulic depth. The 2-, 25-, and 100-year flood discharges were esti- mated on the basis of information from flood- insurance studies of Summers County, Fayette County, and the city of Hinton, and flood-frequency analysis of discharge records for the USGS streamflow-gaging stations at Hinton and Thurmond. The 100-year discharge ranged from 107,000 cubic feet per second at Hinton to 150,000 cubic feet per second at Fayette.

  13. Based on records of Three Gorge Telemetric Seismic Network to analyze Vibration process of micro fracture of rock landslide

    NASA Astrophysics Data System (ADS)

    WANG, Q.

    2017-12-01

    Used the finite element analysis software GeoStudio to establish vibration analysis model of Qianjiangping landslide, which locates at the Three Gorges Reservoir area. In QUAKE/W module, we chosen proper Dynamic elasticity modulus and Poisson's ratio of soil layer and rock stratum. When loading, we selected the waveform data record of Three Gorge Telemetric Seismic Network as input ground motion, which includes five rupture events recorded of Lujiashan seismic station. In dynamic simulating, we mainly focused on sliding process when the earthquake date record was applied. The simulation result shows that Qianjiangping landslide wasn't not only affected by its own static force, but also experienced the dynamic process of micro fracture-creep-slip rupture-creep-slip.it provides a new approach for the early warning feasibility of rock landslide in future research.

  14. Passive seismic experiment in the Olduvai Gorge and Laetoli region (Ngorongoro Conservation Area), Northern Tanzania.

    NASA Astrophysics Data System (ADS)

    Parisi, Laura; Lombardo, Luigi; Tang, Zheng; Mai, P. Martin

    2017-04-01

    The Olduvai Gorge and Laetoli basins, located within the Ngorogoro Conservation Area (NCA), are a cornerstone for understanding the evolution of early humans and are two paleo-antropological excavation sites of global importance. NCA is located at the boundary between the Tanzanian Craton and East African Rift (EAR), in the vicinity of Ngorongoro Crater and other major volcanic edifices. Thus, understanding the geology and tectonics of the NCA may shed light onto the question why early Hominins settled in this region. Environmental and geological conditions in the Olduvai and Laetoli region that promoted human settlement and development are still debated by geologists and paleo-anthropologists. Paleo-geographical reconstructions of the study area of the last 2 million years may take advantage of modern passive seismology. Therefore, we installed a dense seismic network covering a surface of approximately 30 x 40 km within the NCA to map the depth extent of known faults, and to identify seismically active faults that have no surface expression. Our ten seismic stations, equipped with Trillium Compact 120 s sensors, started to operate in June 2016 and will continue for a total of 2 years. At the end of the first year, other 5 stations will densify our network. Here we analyse data quality of the first four months of continuous recordings. Our network provides good quality 3-C waveforms in the frequency range of 0.7-50 Hz. Vertical component seismograms record frequencies reliably down to 8 mHz. Preliminary results of the seismicity obtained with standard location procedures show that NCA is characterised by frequent tectonic seismicity (not volcano-related) with Ml between 0.5 and 2.0. Seismic activity is more frequent in the South (Laetoli region) where major fault systems have not been recognised at the surface yet.

  15. Effects of future sulfate and nitrate deposition scenarios on Linville Gorge and Shining Rock Wildernesses

    Treesearch

    Katherine J. Elliott; James M. Vose; William A. Jackson

    2013-01-01

    We used the Nutrient Cycling Model (NuCM) to simulate the effects of various sulfur (S) and nitrogen (N) deposition scenarios on wilderness areas in Western North Carolina. Linville Gorge Wilderness (LGW) and Shining Rock Wilderness (SRW) were chosen because they are high elevation acidic cove forests and are located on geologic parent material known to be low in base...

  16. Changing landscape in the Three Gorges Reservoir Area of Yangtze River from 1977 to 2005: Land use/land cover, vegetation cover changes estimated using multi-source satellite data

    NASA Astrophysics Data System (ADS)

    Zhang, Jixian; Zhengjun, Liu; Xiaoxia, Sun

    2009-12-01

    The eco-environment in the Three Gorges Reservoir Area (TGRA) in China has received much attention due to the construction of the Three Gorges Hydropower Station. Land use/land cover changes (LUCC) are a major cause of ecological environmental changes. In this paper, the spatial landscape dynamics from 1978 to 2005 in this area are monitored and recent changes are analyzed, using the Landsat TM (MSS) images of 1978, 1988, 1995, 2000 and 2005. Vegetation cover fractions for a vegetation cover analysis are retrieved from MODIS/Terra imagery from 2000 to 2006, being the period before and after the rising water level of the reservoir. Several analytical indices have been used to analyze spatial and temporal changes. Results indicate that cropland, woodland, and grassland areas reduced continuously over the past 30 years, while river and built-up area increased by 2.79% and 4.45% from 2000 to 2005, respectively. The built-up area increased at the cost of decreased cropland, woodland and grassland. The vegetation cover fraction increased slightly. We conclude that significant changes in land use/land cover have occurred in the Three Gorges Reservoir Area. The main cause is a continuous economic and urban/rural development, followed by environmental management policies after construction of the Three Gorges Dam.

  17. [Distribution and risk assessment of mercury species in soil of the water-level-fluctuating zone in the Three Gorges Reservoir].

    PubMed

    Zhang, Cheng; Chen, Hong; Wang, Ding-Yong; Sun, Rong-Guo; Zhang, Jin-Yang

    2014-03-01

    To investigate pollution level and ecological risk of mercury in soils of the water-level-fluctuating zone in the Three Gorges Reservoir Region, 192 surface soil samples from 14 counties (districts) in Chongqing were obtained. Concentrations of THg and Hg species, bioavailable Hg were analyzed and discussed. Geoaccumulation index (I(geo)) and Håkanson potential ecological risk index (E(r)) were applied to assess the pollution status and potential ecological risk of THg and Hg species, respectively. The results showed that significant differences in the concentration of THg were found in soils of water-level-fluctuating zone in the Three Gorges Reservoir. The THg concentration ranged from 22.4 to 393.5 microg x kg(-1), with an average of (84.2 +/- 54.3) microg x kg(-1). 76.6% of the samples' THg content was higher than the soil background value in the Three Gorges Reservoir Region. The percentage of five mercury species (water-soluble Hg, HCl-soluble Hg, KOH-soluble Hg, H2O2-soluble Hg, residue Hg) in soils were 4.1%, 15.5%, 18.3%, 10.9%, 51.3%, respectively. The average concentrations of bioavailable mercury varied between 19.7-36.6 microg x kg(-1), and the percentage of bioavailable Hg was 22.1%-51.6% of THg. According to the geoaccumulation index, the soils were lightly polluted by Hg. Håkanson single potential ecological risk index evaluation showed that Hg species had a low potential ecological risk, moreover, soils of water-level-fluctuating zone in the Three Gorges Reservoir were at low ecological risk levels as evaluated by bioavailable Hg. While, the assessment results based on THg of soils was much higher than that based on the Hg species. Two methods of evaluation showed that the I(geo) and E(r) values calculated based on the Hg species better reflected the actual pollution levels of soils and its hazard to aquatic organisms.

  18. [Nitrogen Losses Under the Action of Different Land Use Types of Small Catchment in Three Gorges Region].

    PubMed

    Chen, Cheng-long; Gao, Ming; Ni, Jiu-pai; Xie, De-ti; Deng, Hua

    2016-05-15

    As an independent water-collecting area, small catchment is the source of non-point source pollution in Three Gorges Region. Choosing 3 kinds of the most representative land-use types and using them to lay monitoring points of overland runoff within the small catchment of Wangjiagou in Fuling of Three Gorges Region, the author used the samples of surface runoff collected through the twelve natural rainfalls from May to December to analyze the feature of spatial-temporal change of Nitrogen's losses concentrations under the influence of different land use types and the hillslopes and small catchments composed by those land use types, revealing the relation between different land-use types and Nitrogen's losses of small catchments in Three Gorges Region. The result showed: the average losses concentration of TN showed the biggest difference for different land use types during the period of spring crops, and the average value of dry land was 1. 61 times and 6.73 times of the values of interplanting field of mulberry and paddy field, respectively; the change of the losses concentration of TN was most conspicuous in the 3 periods of paddy field. The main element was NO₃⁻-N, and the relation between TN and NO₃⁻-N showed a significant linear correlation. TN's and NO₃⁻-N's losses concentrations were significantly and positively correlated with the area ratio of corn and mustard, but got a significant negative correlation with the area ratio of paddy and mulberry; NH₄⁺-N's losses concentrations got a significant positive correlation with the area ratio of mustard. Among all the hillslopes composed by different land use types, TN's average losses concentration of surface runoff of the hillslope composed by interplantating field of mulberry and paddy land during the three periods was the lowest, and the values were 2.55, 11.52, 8.58 mg · L⁻¹, respectively; the hillslope of rotation plough land of corn and mustard had the maximum value, and the values were

  19. Tephrochronology of Bed II, Olduvai Gorge, Tanzania, and placement of the Oldowan-Acheulean transition.

    PubMed

    McHenry, Lindsay J; Stanistreet, Ian G

    2018-04-12

    Tuffaceous marker beds, derived from volcanic products from the Ngorongoro Volcanic Highlands, help define a stratigraphic framework for the world-renowned fossil and stone tool record exposed at Olduvai Gorge, Tanzania. However, previous efforts to constrain this tuff record, especially for Olduvai Bed II, have been limited because of erosion, contamination, reworking, and the alteration of volcanic glass under saline-alkaline conditions. This paper applies previously defined geochemical and mineralogical "fingerprints" for several major Bed II marker tuffs, based on glass (where available) and phenocrysts more resistant to alteration (feldspar, hornblende, augite, and titanomagnetite), to tuffs from stratigraphic sections in the Olduvai Junction Area, including previously and recently excavated Acheulean and Oldowan sites (HWK EE (Locality (Loc) 42), EF-HR (Loc 12a), FLK (Loc 45), and MNK (Loc 88)). The Middle Bed II Bird Print Tuff (BPT) is found to be more compositionally variable than previously reported but is still valuable as a stratigraphic marker over short distances. The confirmation of blocks of Tuff IID in conglomerate helps constrain Upper Bed II stratigraphy at sites where in-situ tuffs are absent. This paper also compiles the results of published geochronological research, providing stratigraphic context and updating previously reported dates using a consistent 40 Ar/ 39 Ar reference standard age. The results of this work support the following paleoanthropologically relevant conclusions: 1) the early Acheulean site EF-HR (Loc 12a) is situated above the level of Hay's Tuff IIC, and thus sits in Upper rather than Middle Bed II, (2) the HWK EE (Loc 42) Oldowan site is constrained between Tuff IIA and Tuff IIB, just above the boundary between Lower and Middle Bed II, and 3) the Acheulean site at FLK W most likely lies within the Middle Augitic Sandstone, above Tuff IIB, similar to the placements by Leakey and Hay for the earliest Acheulean at Olduvai

  20. Design and application analysis of prediction system of geo-hazards based on GIS in the Three Gorges Reservoir

    NASA Astrophysics Data System (ADS)

    Li, Deying; Yin, Kunlong; Gao, Huaxi; Liu, Changchun

    2009-10-01

    Although the project of the Three Gorges Dam across the Yangtze River in China can utilize this huge potential source of hydroelectric power, and eliminate the loss of life and damage by flood, it also causes environmental problems due to the big rise and fluctuation of the water, such as geo-hazards. In order to prevent and predict geo-hazards, the establishment of prediction system of geo-hazards is very necessary. In order to implement functions of hazard prediction of regional and urban geo-hazard, single geo-hazard prediction, prediction of landslide surge and risk evaluation, logical layers of the system consist of data capturing layer, data manipulation and processing layer, analysis and application layer, and information publication layer. Due to the existence of multi-source spatial data, the research on the multi-source transformation and fusion data should be carried on in the paper. Its applicability of the system was testified on the spatial prediction of landslide hazard through spatial analysis of GIS in which information value method have been applied aims to identify susceptible areas that are possible to future landslide, on the basis of historical record of past landslide, terrain parameter, geology, rainfall and anthropogenic activity. Detailed discussion was carried out on spatial distribution characteristics of landslide hazard in the new town of Badong. These results can be used for risk evaluation. The system can be implemented as an early-warning and emergency management tool by the relevant authorities of the Three Gorges Reservoir in the future.

  1. Assessing potential release tendency of As, Mo and W in the tributary sediments of the Three Gorges Reservoir, China.

    PubMed

    Gao, Li; Gao, Bo; Peng, Wenqi; Xu, Dongyu; Yin, Shuhua

    2018-01-01

    As the largest man-made reservoir in China, the Three Gorges Reservoir (TGR) has significant influence on national drinking water safety. The geochemical behavior of trace elements at the sediment-water interface (SWI) is still unknown. The mobilization characteristics of trace elements (As, Mo and W)-determined by diffusive gradients in thin films (DGT)-were studied to quantitatively calculate the release trends in the SWI in three typical tributaries and the mainstream of the TGR in the summer. The results showed that concentrations of DGT-labile As, Mo and W in the overlying water and sediment cores showed significant variations in the ranges of 0.05-50.90, 0.30-1.63 and 0.01-0.42μgL -1 , respectively. The apparent net diffusive fluxes were significantly positive in most sampling sites (77.8% for As, 88.8% for Mo and 66.6% for W), suggesting that the sediment was the source of these three elements. It was noteworthy that the maximum net diffusive fluxes of As and W were found in the upstream of Meixi tributary, which may be attributed to anthropogenic activities. In addition, As, Mo and W may be incorporated in Fe and Mn oxyhydroxides and these three elements simultaneously remobilized with Fe and Mn. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Crustal structure across the Three Gorges area of the Yangtze platform, central China, from seismic refraction/wide-angle reflection data

    USGS Publications Warehouse

    Zhang, Z.; Bai, Z.; Mooney, W.; Wang, C.; Chen, X.; Wang, E.; Teng, J.; Okaya, N.

    2009-01-01

    We present active-source seismic data recorded along a 300??km-long profile across the Three Gorges area of the western Yangtze platform, central China. From west to east, the profile crosses the Zigui basin, Huangling dome and Jianghan basin. The derived crustal P-wave velocity structure changes significantly across the Tongchenghe fault that lies at the transition from the Huangling dome to the Jianghan basin. West of the Tongchenghe fault, beneath the Zigui basin and the Huangling dome, we observe a ~ 42??km thick crust of relatively low average velocity (6.3-6.4??km/s). In contrast, east of the Tongchenghe fault, beneath the Jianghan basin, the crust is only 30??km thick and has a high average velocity (6.6-6.7??km/s). A west-east variation in crustal composition along the Tongchenghe fault is also inferred. West of the fault, P-wave velocities suggest a felsic composition with an intermediate layer at the base of the crust, whilst, east of the fault, felsic, intermediate, and mafic crustal layers are apparent. Our results suggest that the crust beneath the Jianghan basin has been thinned by rifting, accompanied by intrusion of the lower crust by mafic dikes and sills. The west-to-east division of the crust in the Three Gorges area coincides with first-order geophysical contrasts in gravity, topography, crustal and lithospheric thickness. ?? 2009 Elsevier B.V.

  3. [Spatio-temporal changes of nitrogen balance in 1980-2005 for agricultural land in Three Gorges Reservoir Area].

    PubMed

    Xu, Xi-bao; Yang, Gui-shan; Li, Heng-peng

    2009-08-15

    Based on the long-term agricultural statistics data at the county scale, the estimation of nitrogen balance from 1980 to 2005 for agricultural land in Three Gorges Reservoir Area was made by the OECD soil surface nitrogen balance model with some suitable modification. The spatio-temporal changes of nitrogen balance and its drivers were analyzed. The results showed that the total inputs and total surplus of nitrogen from 1980 to 2005 presented increasing trends continuously, from 23.4 x 10(4) t and 14.4 x 104 t to 45.6 x 10(4) t and 30 x 10(4) t respectively. The total output of nitrogen in 1980-1995 was at the increasing trend, from 9.0 x 10(4) t to 16.7 x 10(4) t, while that of 1996-2005 was keeping steady. The average unit surplus of nitrogen in 1980-1998 was also at the increasing trend, from 133.4 kg/hm2 to 310.3 kg/hm(2); and the trend inclined to be steady after 1998, while the spatial differential pattern toned up. The great spatial changes for nitrogen surplus from 1980 to 2005, mainly centralized at the head and the middle of the Three Gorges Reservoir Area, similar to the spatial distribution of the resettlement. Fertilizer, manure and biological fixation were the main contributors of nitrogen input sources, accumulatively totaled for above 90%. Nitrogen balance changes were mainly influenced by the macro-environment of fertilizer utilization before 1995, while which were influenced by the large amounts of the resettlement for Three Gorges Project after 1995. However, how much the effects of the resettlement on nitrogen balance need to be further explored. Developing sideline, agricultural structure transition or ecological resettlement should be considered to control nitrogen emission.

  4. The German-Chinese research collaboration YANGTZE-GEO: Assessing the geo-risks in the Three Gorges Reservoir area

    NASA Astrophysics Data System (ADS)

    Schönbrodt, S.; Behrens, T.; Bieger, K.; Ehret, D.; Frei, M.; Hörmann, G.; Seeber, C.; Schleier, M.; Schmalz, B.; Fohrer, N.; Kaufmann, H.; King, L.; Rohn, J.; Subklew, G.; Xiang, W.

    2012-04-01

    The river impoundment by The Three Gorges Dam leads to resettlement and land reclamation on steep slopes. As a consequence, ecosystem changes such as soil erosion, mass movements, and diffuse sediment and matter fluxes are widely expected to increase rapidly. In order to assess and analyse those ecosystem changes, the German-Chinese joint research project YANGTZE-GEO was set up in 2008. Within the framework of YANGTZE-GEO five German universities (Tuebingen, Erlangen, Giessen, Kiel, Potsdam) conducted studies on soil erosion, mass movements, diffuse matter inputs, and land use change and vulnerability in close collaboration with Chinese scientists. The Chinese partners and institutions are according to their alphabetic order of hometown the Chinese Research Academy of Environmental Sciences (CRAES; Beijing), the Standing Office of the State Council Three Gorges Project Construction Committee (Beijing), the National Climate Centre (NCC) of the China Meteorological Administration (CMA; Beijing), the Aero Geophysical Survey and Remote Sensing for Land and Resources (AES; Beijing), the Nanjing University, the CAS Institute of Soil Science (Nanjing), the Nanjing Institute of Geography and Limnology at CAS (NIGLAS; Nanjing), the China University of Geosciences (CUG; Wuhan), the CAS Institute of Hydrobiology (Wuhan), and the China Three Gorges University (Yichang). The overall aim of YANGTZE-GEO is the development of a risk assessment and forecasting system to locate high risk areas using GIS-based erosion modelling, data mining tools for terrace condition analysis and landslide recognition, eco-hydrological modelling for diffuse matter inputs, and state-of-the-art remote sensing to assess the landscape's vulnerability. Furthermore, the project aims at the recommendation of sustainable land management systems. YANGTZE-GEO showed the relevance of such research and crucially contributes to the understanding of the dimension and dynamics of the ecological consequences of

  5. Human activity and landscape change at Adjiyska Vodenitsa, central Bulgaria

    NASA Astrophysics Data System (ADS)

    Chiverrell, R. C.; Archibald, Z.

    2009-04-01

    The Classical and early Hellenistic settlement located at Adjiyska Vodenitsa, near Vetren, in the centre of ancient Thrace, is somewhat unusual in displaying well-preserved evidence for the commercial and cultural interactions associated with a river port. The settlement evolved through a series of phases, with traces of activity emerging around the beginning of the fifth, advancing to a progressive close in the second century BCE. The settlement at Adjiyska Vodenitsa owed its raison d'être to river traffic, with the river as the primary means for importing commodities (non-local amphorae and roof tiles). The Pistiros inscription indicates that overland traffic was also important, presumably over the Rhodopes Mountains to the south. The relationship between the site and the Maritsa River is thus important. Located towards the head of the Maritsa basin downstream of the Momina Klissoura gorge near Belovo, the settlement is perched on a steeply-edged toe of the Vetren tributary alluvial fan, with the bluff trimmed by the migrating River Maritsa. The site is c.12-14 metres above the current bed of the River Maritsa, high and dry away from contemporary flooding, intriguing given evidence for flooding during occupation. The fluvial system has clearly been dynamic and changed much during the last 3000 years. Changes in the Maritsa have been constrained with radiocarbon ages obtained for two fan terrace levels. Active channel and overbank environments of the higher B1 terrace are dated to c. 520-400 cal. BC and provide a terminus post quem for the incision to the lower terrace (B2). The radiocarbon dating of basal contexts from the lower terrace palaeo-channels provides a terminus ante quem for abandonment of the higher terrace of cal. AD 1010-1150. Thus basal lowering and a shift to greater lateral channel activity appear to coincide with the abandonment of Adjiyska Vodenitsa. Upstream of the Momina Klissoura Gorge evidence for heightened geomorphic activity probably

  6. The Empirical Study on the Labor Export of Three Gorges Reservoir Area in China

    NASA Astrophysics Data System (ADS)

    Hu, Bangyong

    There are many large surplus labor force in the three gorges reservoir area, export of labor services is one way to tackle the problem of employment of the surplus-labor and increase farmers income, export of labor is also a effective way to solve three rural issues. This paper analyzes the need for the development of service economy, study the problems of export of labor services, at last the author give some suggestion to develop labor economy.

  7. [Distribution of Mercury in Plants at Water-Level-Fluctuating Zone in the Three Gorges Reservoir].

    PubMed

    Liang, Li; Wang, Yong-min; Li, Xian-yuan; Tang, Zhen-ya; Zhang, Xiang; Zhang, Cheng; WANG, Ding-yong

    2015-11-01

    The mercury (Hg) distribution and storage in plants at water-level-fluctuating zone (WLFZ) in the Three Gorges Reservoir were investigated by analyzing the total mercury(THg) and methylmercury ( MeHg) levels in different parts of plants collected from three typical sites including Shibaozhai, Zhenxi and Hanfeng Lake in WLFZ. The results indicated that THg and MeHg concentrations in plants ranged from (1.62 ± 0.57) to (49.42 ± 3.93) μg x kg(-1) and from (15.27 ± 7.09) to (1 974.67 ± 946.10) ng x kg(-1), respectively. In addition, THg levels in different plant parts followed the trend: root > leaf > stem, and similar trend for MeHg was observed with the highest level in root. An obvious spatial distribution was also found with the THg and MeHg levels in plants in Hanfeng higher than those in the same plants in the other two sampling sites (Shibaozhai and Zhenxi), and there was a difference of THg and MeHg storage in plants in various attitudes. The corresponding THg and MeHg storages were 145.3, 166.4, 124.3 and 88.2 mg x hm(-2), and 1.9, 2.7, 3.6 and 3.2 mg x hm(-2) in 145-150, 150-160, 160-170 and 170-175 m attitudes. The accumulation ability of dominant plants in WLFZ for THg (bioaccumulation factor, BAF < 1) was weaker than that for MeHg (BAF > 1).

  8. Mineral resource potential maps of the Fiddler Butte Wilderness Study Area and the Fremont Gorge Study Area, Garfield and Wayne counties, Utah

    USGS Publications Warehouse

    Larson, M.J.; Dubiel, R.F.; Peterson, Fred; Willson, W.R.; Briggs, J.P.

    1985-01-01

    Field and laboratory investigations of the Fiddler Butte WSA (Wilderness Study Area) in Garfield County, Utah, and of the Fremont Gorge study area in Wayne County, Utah, were made to determine the mineral resource potential of these lands. The investigations indicate that two areas in the northeastern and southwestern parts of the Fiddler Butte WSA have a moderate potential for uranium resources. The entire Fiddler Butte WSA has a moderate potential for petroleum resources, and the northeastern part of the WSA has a high potential for tar sand resources. The studies indicate a low potential for metallic and nonmetallic resources in the Fiddler Butte WSA. The Fremont Gorge study area has a low potential for metallic, nonmetallic, and petroleum resources.

  9. Normal Modes Expose Active Sites in Enzymes.

    PubMed

    Glantz-Gashai, Yitav; Meirson, Tomer; Samson, Abraham O

    2016-12-01

    Accurate prediction of active sites is an important tool in bioinformatics. Here we present an improved structure based technique to expose active sites that is based on large changes of solvent accessibility accompanying normal mode dynamics. The technique which detects EXPOsure of active SITes through normal modEs is named EXPOSITE. The technique is trained using a small 133 enzyme dataset and tested using a large 845 enzyme dataset, both with known active site residues. EXPOSITE is also tested in a benchmark protein ligand dataset (PLD) comprising 48 proteins with and without bound ligands. EXPOSITE is shown to successfully locate the active site in most instances, and is found to be more accurate than other structure-based techniques. Interestingly, in several instances, the active site does not correspond to the largest pocket. EXPOSITE is advantageous due to its high precision and paves the way for structure based prediction of active site in enzymes.

  10. Normal Modes Expose Active Sites in Enzymes

    PubMed Central

    Glantz-Gashai, Yitav; Samson, Abraham O.

    2016-01-01

    Accurate prediction of active sites is an important tool in bioinformatics. Here we present an improved structure based technique to expose active sites that is based on large changes of solvent accessibility accompanying normal mode dynamics. The technique which detects EXPOsure of active SITes through normal modEs is named EXPOSITE. The technique is trained using a small 133 enzyme dataset and tested using a large 845 enzyme dataset, both with known active site residues. EXPOSITE is also tested in a benchmark protein ligand dataset (PLD) comprising 48 proteins with and without bound ligands. EXPOSITE is shown to successfully locate the active site in most instances, and is found to be more accurate than other structure-based techniques. Interestingly, in several instances, the active site does not correspond to the largest pocket. EXPOSITE is advantageous due to its high precision and paves the way for structure based prediction of active site in enzymes. PMID:28002427

  11. Effects of land use patterns on stream water quality: a case study of a small-scale watershed in the Three Gorges Reservoir Area, China.

    PubMed

    Huang, Zhilin; Han, Liyang; Zeng, Lixiong; Xiao, Wenfa; Tian, Yaowu

    2016-02-01

    In this study, we have considered the relationship between the spatial configuration of land use and water quality in the Three Gorges Reservoir Area. Using land use types, landscape metrics, and long-term water quality data, as well as statistical and spatial analysis, we determined that most water quality parameters were negatively correlated with non-wood forest and urban areas but were strongly positively correlated with the proportion of forest area. Landscape indices such as patch density, contagion, and the Shannon diversity index were able to predict some water quality indicators, but the mean shape index was not significantly related to the proportions of farmland and water in the study area. Regression relationships were stronger in spring and fall than in summer, and relationships with nitrogen were stronger than those of the other water quality parameters (R(2) > 0.80) in all three seasons. Redundancy analysis showed that declining stream water quality was closely associated with configurations of urban, agricultural, and forest areas and with landscape fragmentation (PD) caused by urbanization and agricultural activities. Thus, a rational land use plan of adjusting the land use type, controlling landscape fragmentation, and increasing the proportion of forest area would help to achieve a healthier river ecosystem in the Three Gorges Reservoir Area (TGRA).

  12. The Hydrological Regimes Brought by the Three Gorges Project Affected Riparian Vegetation Distribution and Diversity in 2009 and 2010

    NASA Astrophysics Data System (ADS)

    Miao, Ling-Feng; Liu, Wei-Wei; Yang, Fan

    2017-01-01

    Post-dam riparian vegetations affected by the new hydrological regimes in the Three Gorges Reservoir (TGR) were investigated in 2009 and 2010, respectively. The investigation in 2009 showed that about 231 vascular plant species belonging to 169 genera of 61 families were distributed in the water-level-fluctuation zone (WLFZ) of the (TGR). Three vegetation types, including Chuanjiang, Gorge, and other vegetation types, were classified efficiently via cluster analysis. Alpha diversity analysis indicated that species richness gradually decreased with decreasing elevation. Beta diversity analysis indicated that high environment heterogeneity was existed between the lower section and the other two sections, and environment homogeneity was also existed between middle section and upper section. Using the analysis of the field growth in the 2009 and 2010 field surveys as bases, we proposed a list of perennial herb species and woody species that may potentially occurred in the WLFZ of the TGR. In addition, we predicted plant community structural changes in the different altitude sections of WLFZ in the future.

  13. Study on Seismogenesis of 2013 Ms5.1 Badong Earthquake in the Three Gorges Reservoir Region

    NASA Astrophysics Data System (ADS)

    Li, X.; Zeng, Z.; Xu, S.; He, C.

    2015-12-01

    On 16 December, 2013, an earthquake of Ms5.1 occurred in Badong County, the Three Gorges Reservoir area, China. We collected all the 150 published focal mechanism solutions (FMS) and inversed the tectonic stress field in Badong, the Three Gorges Dam and Huangling anticline area using the software SATSI (Hardebeck and Michael, 2006). Inversion results show that the orientations of maximum principle stress axis (σ1) in Badong plunge to NNE or SSW. Detailed characteristics of the stress field indicate that the σ1 axis is almost vertical in the center of Huangling anticline and turns horizontal to the west. As to deep structures, we studied the satellite gravity anomalies of 8-638 order in this area using the EIGEN-6C2 model provided by ICGRM. Combining the seismic sounding profile through the epicenter of Badong earthquake and the petrology data, we reinterpreted the deep structure in the study area. The results show that the deep crust in Badong is unstable and the deep material's upwelling leads to Huangling anticline continued uplifting, which is consistent with the result indicated from the stress filed. Both of them provide energy for the preparation of earthquake. The FMS shows that Gaoqiao Fault is the causative fault of this Ms5.1 earthquake. Field investigations indicated that the lithology and fracture characteristic in Badong is beneficial to reservoir water infiltration. Before the earthquake, reservoir water level raised to 175m, the highest storage level, which increased the loading. Based on above researches, we believe that the Ms5.1 Badong earthquake is controlled by deep tectonic environment and stress field in shallow crust. The reservoir water infiltration and uploading increase generated by water storage of the Three Gorges area reduced the strength of Gaoqiao Fault and changed its stress state. These factors jointly promoted an abrupt movement of the fault in the critical stress state, and triggered the Ms5.1 Badong earthquake.

  14. [Effects of Citric Acid on Activation and Methylation of Mercury in the Soils of Water-Level-Fluctuating Zone of the Three Gorges.Reservoir].

    PubMed

    Qin, Cai-qing; Liang, Li; You, Rui; Deng, Han; Wang, Ding-yong

    2015-12-01

    To investigate effects of the main component of vegetation root exudates-citric acid on activation and methylation of mercury in the soil of water-level-fluctuating zone (WLFZ) of the Three Gorges Reservoir area, simulation experiments were conducted by extracting and cultivating soil with different concentrations of citric acid. The results showed that after adding citric acid, the total mercury content in leaching solution before reaching peak were higher than that of the control, and increased with the increase of citric acid concentrations. The maximum amount of mercury complexes increased initially and then reached plateaus with the percentage against the total mercury in soil of 1.03%, 1.67%, 1.99%, 2.47%, 2.68%, 2.73% and 2.73% for different citric acid concentrations (0, 1, 2, 4, 5, 6 and 8 mmol · L⁻¹). In addition, concentrations of methylmercury ( MeHg) in soil remained stable in the first 3 hours, and then increased accompanying with the increasing rate rising with the concentration of citric acid ( besides the control group) . This result indicated that citric acid probably could promote the transformation process from inorganic mercury to MeHg in soil. which increased with the concentration of citric acid.

  15. Biostable insect kinin analogs reduce blood meal and disrupt ecdysis in the blood-gorging Chagas’ disease vector, Rhodnius prolixus

    USDA-ARS?s Scientific Manuscript database

    Rhodnius prolixus is a blood-gorging hemipteran that takes blood meals that are approximately 10 times its body weight. This blood meal is crucial for growth and development and is needed to ensure a successful molt into the next instar. Kinins are a multifunctional family of neuropeptides which hav...

  16. Quantifying Oldowan Stone Tool Production at Olduvai Gorge, Tanzania

    PubMed Central

    Reti, Jay S.

    2016-01-01

    Recent research suggests that variation exists among and between Oldowan stone tool assemblages. Oldowan variation might represent differential constraints on raw materials used to produce these stone implements. Alternatively, variation among Oldowan assemblages could represent different methods that Oldowan producing hominins utilized to produce these lithic implements. Identifying differential patterns of stone tool production within the Oldowan has implications for assessing how stone tool technology evolved, how traditions of lithic production might have been culturally transmitted, and for defining the timing and scope of these evolutionary events. At present there is no null model to predict what morphological variation in the Oldowan should look like. Without such a model, quantifying whether Oldowan assemblages vary due to raw material constraints or whether they vary due to differences in production technique is not possible. This research establishes a null model for Oldowan lithic artifact morphological variation. To establish these expectations this research 1) models the expected range of variation through large scale reduction experiments, 2) develops an algorithm to categorize archaeological flakes based on how they are produced, and 3) statistically assesses the methods of production behavior used by Oldowan producing hominins at the site of DK from Olduvai Gorge, Tanzania via the experimental model. Results indicate that a subset of quartzite flakes deviate from the null expectations in a manner that demonstrates efficiency in flake manufacture, while some basalt flakes deviate from null expectations in a manner that demonstrates inefficiency in flake manufacture. The simultaneous presence of efficiency in stone tool production for one raw material (quartzite) and inefficiency in stone tool production for another raw material (basalt) suggests that Oldowan producing hominins at DK were able to mediate the economic costs associated with stone tool

  17. Amphitheater-headed canyons formed by megaflooding at Malad Gorge, Idaho

    PubMed Central

    Lamb, Michael P.; Mackey, Benjamin H.; Farley, Kenneth A.

    2014-01-01

    Many bedrock canyons on Earth and Mars were eroded by upstream propagating headwalls, and a prominent goal in geomorphology and planetary science is to determine formation processes from canyon morphology. A diagnostic link between process and form remains highly controversial, however, and field investigations that isolate controls on canyon morphology are needed. Here we investigate the origin of Malad Gorge, Idaho, a canyon system cut into basalt with three remarkably distinct heads: two with amphitheater headwalls and the third housing the active Wood River and ending in a 7% grade knickzone. Scoured rims of the headwalls, relict plunge pools, sediment-transport constraints, and cosmogenic (3He) exposure ages indicate formation of the amphitheater-headed canyons by large-scale flooding ∼46 ka, coeval with formation of Box Canyon 18 km to the south as well as the eruption of McKinney Butte Basalt, suggesting widespread canyon formation following lava-flow diversion of the paleo-Wood River. Exposure ages within the knickzone-headed canyon indicate progressive upstream younging of strath terraces and a knickzone propagation rate of 2.5 cm/y over at least the past 33 ka. Results point to a potential diagnostic link between vertical amphitheater headwalls in basalt and rapid erosion during megaflooding due to the onset of block toppling, rather than previous interpretations of seepage erosion, with implications for quantifying the early hydrosphere of Mars. PMID:24344293

  18. Will river erosion below the Three Gorges Dam stop in the middle Yangtze?

    NASA Astrophysics Data System (ADS)

    Lai, X.; Yin, D.; Finlayson, B. L.; Wei, T.; Li, M.; Yuan, W.; Yang, S.; Dai, Z.; Gao, S.; Chen, Z.

    2017-11-01

    The environmental impact of the Three Gorges Dam has been a subject of vigorous academic, political and social debate since its inception. This includes the key issue of post-dam river channel erosion, which was predicted by the feasibility study to extend to the river mouth. In this paper we examine the geomorphic response of the channel of the middle Yangtze for 660 km downstream of the dam. Using data on channel characteristics, bed material and sediment transport, we show that in the decade following the dam closure, pre-dam seasonal erosion has been replaced by year-round erosion, a pattern most marked at the upstream end of the study area. The sediment carrying capacity of the river channel has been largely reduced below the dam. The locus of bed scour has moved progressively downstream, ceasing as the bed material became too coarse to be transported (e.g. D50: 0.29 mm pre-dam coarsened to 20 mm below the dam by 2008). About 400 km below the dam there is a reduction in channel slope that changes the sediment carrying capacity from 0.25 kg m-3 to only about 0.05 kg m-3, which is insufficient to move bed sediment. The new long-term hydro-morphological equilibrium that will be established in this section of the middle Yangtze will prevent the further incision downstream initiated by the Three Gorges Dam. The results suggest that the full extent of adverse environmental impact predicted by the pre-dam studies will not eventuate.

  19. Channel evolution under changing hydrological regimes in anabranching reaches downstream of the Three Gorges Dam

    NASA Astrophysics Data System (ADS)

    Han, Jianqiao; Zhang, Wei; Yuan, Jing; Fan, Yongyang

    2018-03-01

    Elucidating the influence of dams on fluvial processes can benefit river protection and basin management. Based on hydrological and topographical data, we analyzed channel evolution in anabranching reaches under changing hydrological regimes influenced by the Three Gorges Dam. The main conclusions are as follows: 1) the channels of specific anabranching reaches were defined as flood trend channels or low-flow trend channels according to the distribution of their flow characteristics. The anabranching reaches were classified as T1 or T2. The former is characterized by the correspondence between the flood trend and branch channels, and the latter is characterized by the correspondence between the flood trend and main channels; 2) on the basis of the new classification, the discrepant patterns of channel evolution seen in anabranching reaches were unified into a pattern that showed flood trend channels shrinking and low-flow trend channels expanding; 3) flood abatement and the increased duration of moderate flow discharges are the main factors that affect channel adjustments in anabranching reaches after dam construction; and 4) in the next few decades, the pattern of channel evolution will remain the same as that of the Three Gorges Dam operation. That is, the morphology will fully adapt to a flow with a low coefficient of variation. Our results are of interest in the management of the Yangtze River and other rivers influenced by dams.

  20. Assessment of online public opinions on large infrastructure projects: A case study of the Three Gorges Project in China

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jiang, Hanchen, E-mail: jhc13@mails.tsinghua.edu.cn; Qiang, Maoshan, E-mail: qiangms@tsinghua.edu.cn; Lin, Peng, E-mail: celinpe@mail.tsinghua.edu.cn

    Public opinion becomes increasingly salient in the ex post evaluation stage of large infrastructure projects which have significant impacts to the environment and the society. However, traditional survey methods are inefficient in collection and assessment of the public opinion due to its large quantity and diversity. Recently, Social media platforms provide a rich data source for monitoring and assessing the public opinion on controversial infrastructure projects. This paper proposes an assessment framework to transform unstructured online public opinions on large infrastructure projects into sentimental and topical indicators for enhancing practices of ex post evaluation and public participation. The framework usesmore » web crawlers to collect online comments related to a large infrastructure project and employs two natural language processing technologies, including sentiment analysis and topic modeling, with spatio-temporal analysis, to transform these comments into indicators for assessing online public opinion on the project. Based on the framework, we investigate the online public opinion of the Three Gorges Project on China's largest microblogging site, namely, Weibo. Assessment results present spatial-temporal distributions of post intensity and sentiment polarity, reveals major topics with different sentiments and summarizes managerial implications, for ex post evaluation of the world's largest hydropower project. The proposed assessment framework is expected to be widely applied as a methodological strategy to assess public opinion in the ex post evaluation stage of large infrastructure projects. - Highlights: • We developed a framework to assess online public opinion on large infrastructure projects with environmental impacts. • Indicators were built to assess post intensity, sentiment polarity and major topics of the public opinion. • We took the Three Gorges Project (TGP) as an example to demonstrate the effectiveness proposed framework.

  1. Effects of wildfires and liming of pine-oak-heath communities in the Linville Gorge Wilderness, western North Carolina

    Treesearch

    Katherine J. Elliott; James M. Vose; Jennifer D. Knoepp; William Jackson

    2012-01-01

    Linville Gorge Wilderness (LGW) is a Class I area in the southern Appalachian Mountains, western North Carolina. Over the last 150 years, LGW has been subject to several wildfires, varying in intensity and extent (Newell and Peet 1995). In November 2000, a wildfire burned 4000 ha in the wilderness; the fire ranged in severity across the northern portion of the...

  2. Ten years of the Three Gorges Dam: a call for policy overhaul

    NASA Astrophysics Data System (ADS)

    Yang, Xiankun; Lu, X. X.

    2013-12-01

    The Three Gorges Dam (TGD), the world’s largest source of ‘clean’ hydroelectric power (Shen and Xie 2004), has entered its tenth year after the first turbine went into operation in June 2003. The dam, with a generating capacity 20 times that of the United States’ Hoover Dam, has been hailed as a crucial part of a solution to China’s energy crisis. Despite great benefits, however, major concerns have been voiced over the disastrous environmental and social consequences of this massive engineering project (Stone 2011). In this paper, we review the benefits and impacts learned from the controversial megadam over the past decade and discuss perspective quests on policy overhaul for future environmental protection.

  3. Polycyclic aromatic hydrocarbons in bottom sediment and bioavailability in streams in the New River Gorge National River and Gauley River National Recreation Area, West Virginia, 2002

    USGS Publications Warehouse

    Messinger, Terrence

    2004-01-01

    Polycyclic aromatic hydrocarbons (PAHs), including some on the U.S. Environmental Protection Agency's priority pollutant list, were found in bottom sediment in streams in the coal-producing region of the Kanawha River Basin in 1996-1998, and in and near the New River Gorge National River in 2002, in concentrations exceeding those thought likely to cause adverse effects to wildlife. Very low concentrations of bioavailable PAHs were measured in streams in and near the New River Gorge National River by the use of semipermeable membrane devices. The apparent contradiction between the high concentrations of total PAHs and the low concentrations of bioavailable PAHs may result from the presence of a substantial amount of particulate coal in bottom sediment.

  4. Simulation of land use change in the three gorges reservoir area based on CART-CA

    NASA Astrophysics Data System (ADS)

    Yuan, Min

    2018-05-01

    This study proposes a new method to simulate spatiotemporal complex multiple land uses by using classification and regression tree algorithm (CART) based CA model. In this model, we use classification and regression tree algorithm to calculate land class conversion probability, and combine neighborhood factor, random factor to extract cellular transformation rules. The overall Kappa coefficient is 0.8014 and the overall accuracy is 0.8821 in the land dynamic simulation results of the three gorges reservoir area from 2000 to 2010, and the simulation results are satisfactory.

  5. Distribution and health risk assessment of dissolved heavy metals in the Three Gorges Reservoir, China (section in the main urban area of Chongqing).

    PubMed

    Zhao, Xin; Li, Ting-Yong; Zhang, Tao-Tao; Luo, Wei-Jun; Li, Jun-Yun

    2017-01-01

    The Three Gorges Project (TGP) is the largest hydropower station ever built in the world. A better understanding of the concentrations of heavy metals in the aquatic environment of the Three Gorges Reservoir (TGR) is crucial for national drinking water security and sustainable ecosystem development. To thoroughly investigate the impact of heavy metals on water quality after the impoundment to the maximum level of 175 m in the TGR, the concentrations of the dissolved heavy metals (Cr, Cu, Zn, Cd, Pb, As) were measured in April and August 2015, by inductively coupled plasma mass spectrometry (ICP-MS). (1) Except Zn and Pb, most of the heavy metal concentrations in the water of the TGR reached the level of the National Surface Water Environmental Quality Standards (GB3838-2002) I of China, revealing that the water quality of the TGR was good overall. (2) There were significant positive correlations among the concentrations of Cu, As, and Cd, revealing that they may exhibit similar geochemical behaviors. (3) The spatial distribution of the heavy metal concentrations was diverse and complex. The Zn concentration obviously increased in the rainy season from upstream to downstream in the Yangtze River, while the other heavy metals exhibited no significant changes in their concentrations. The distribution characteristics of the heavy metal concentrations on both sides and the middle of the river were different at different sites. (4) The health risk of the six elements was assessed through a human health risk assessment (HHRA), and the assessment results were lower than the maximum acceptable risk level designed by the US EPA and International Commission on Radiological Protection (ICRP). The HHRA model in the aquatic environment revealed that the risk of non-carcinogenic heavy metals (Cu, Zn, and Pb) was at a negligible risk level of 10 -11 ∼10 -9  a -1 . At all the study sites, the risk of carcinogenic heavy metals (Cr, Cd, and As) was higher than the risk of non

  6. Dissecting the active site of a photoreceptor protein

    NASA Astrophysics Data System (ADS)

    Hoff, Wouter; Hara, Miwa; Ren, Jie; Moghadam, Farzaneh; Xie, Aihua; Kumauchi, Masato

    While enzymes are quite large molecules, functionally important chemical events are often limited to a small region of the protein: the active site. The physical and chemical properties of residues at such active sites are often strongly altered compared to the same groups dissolved in water. Understanding such effects is important for unraveling the mechanisms underlying protein function and for protein engineering, but has proven challenging. Here we report on our ongoing efforts on using photoactive yellow protein (PYP), a bacterial photoreceptor, as a model system for such effects. We will report on the following questions: How many residues affect active site properties? Are these residues in direct physical contact with the active site? Can functionally important residues be recognized in the crystal structure of a protein? What structural resolution is needed to understand active sites? What spectroscopic techniques are most informative? Which weak interactions dominate active site properties?

  7. Active site dynamics of ribonuclease.

    PubMed Central

    Brünger, A T; Brooks, C L; Karplus, M

    1985-01-01

    The stochastic boundary molecular dynamics method is used to study the structure, dynamics, and energetics of the solvated active site of bovine pancreatic ribonuclease A. Simulations of the native enzyme and of the enzyme complexed with the dinucleotide substrate CpA and the transition-state analog uridine vanadate are compared. Structural features and dynamical couplings for ribonuclease residues found in the simulation are consistent with experimental data. Water molecules, most of which are not observed in crystallographic studies, are shown to play an important role in the active site. Hydrogen bonding of residues with water molecules in the free enzyme is found to mimic the substrate-enzyme interactions of residues involved in binding. Networks of water stabilize the cluster of positively charged active site residues. Correlated fluctuations between the uridine vanadate complex and the distant lysine residues are mediated through water and may indicate a possible role for these residues in stabilizing the transition state. Images PMID:3866234

  8. Heavy metal pollution in sediments of the largest reservoir (Three Gorges Reservoir) in China: a review.

    PubMed

    Zhao, Xingjuan; Gao, Bo; Xu, Dongyu; Gao, Li; Yin, Shuhua

    2017-09-01

    The Three Gorges Dam in China is the world's largest dam. Upon its completion in 2003, the Three Gorges Reservoir (TGR) became the largest reservoir in China and plays an important role in economic development and national drinking water safety. However, as a sink and source of heavy metals, there is a lack of continuous and comparative data on heavy metal pollution in sediments. This study reviewed all available literatures published on heavy metals in TGR sediments and further provided a comprehensive assessment of the pollution tendency of these heavy metals. The results showed that heavy metal concentrations in TGR sediments varied spatially and temporally. Temporal variations indicated that Hg in tributaries, as well as As, Cd, Cr, Cu, Ni, Pb, and Zn in the mainstream, exhibited a higher probability to exceed background values after the impoundment of TGR. Pollution assessments by contamination factor, geoaccumulation index, and potential ecological risk were similar. High Cd and Hg concentrations in both the mainstream and tributaries are a cause for much concern. However, sediment quality guidelines produced different results, as most previous studies adopted different sampling and measurement strategies. The data inconsistencies and lack of continuity regarding the reservoir confirm the need for a continuous monitoring network and the development of quality criteria relevant to the sediments of the TGR in the future.

  9. In situ ∼2.0 Ma trees discovered as fossil rooted stumps, lowermost Bed I, Olduvai Gorge, Tanzania.

    PubMed

    Habermann, Jörg M; Stanistreet, Ian G; Stollhofen, Harald; Albert, Rosa M; Bamford, Marion K; Pante, Michael C; Njau, Jackson K; Masao, Fidelis T

    2016-01-01

    The discovery of fossil rooted tree stumps in lowermost Lower Bed I from the western Olduvai Basin, Tanzania, age-bracketed by the Naabi Ignimbrite (2.038 ± 0.005 Ma) and Tuff IA (1.88 ± 0.05 Ma), provides the first direct, in situ, and to date oldest evidence of living trees at Olduvai Gorge. The tree relicts occur in an interval dominated by low-viscosity mass flow and braided fluvial sediments, deposited at the toe of a largely Ngorongoro Volcano-sourced volcaniclastic fan apron that comprised a widely spaced network of ephemeral braided streams draining northward into the Olduvai Basin. Preservation of the trees occurred through their engulfment by mass flows, post-mortem mold formation resulting from differential decay of woody tissues, and subsequent fluvially-related sediment infill, calcite precipitation, and cast formation. Rhizolith preservation was triggered by the interaction of root-induced organic and inorganic processes to form rhizocretionary calcareous root casts. Phytolith analyses were carried out to complete the paleoenvironmental reconstruction. They imply a pronounced seasonality and indicate a wooded landscape with grasses, shrubs, and sedges growing nearby, comparable to the low, open riverine woodland (unit 4c) along the Garusi River and tributaries in the Laetoli area. Among the tree stump cluster were found outsized lithic clasts and those consisting of quartzite were identified as Oldowan stone tool artifacts. In the context of hominin activity, the identification of wooded grassland in association with nearby freshwater drainages and Oldowan artifacts significantly extends our paleoenvironmental purview on the basal parts of Lower Bed I, and highlights the hitherto underrated role of the yet poorly explored western Olduvai Gorge area as a potential ecologically attractive setting and habitat for early hominins. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Low dielectric response in enzyme active site

    PubMed Central

    Mertz, Edward L.; Krishtalik, Lev I.

    2000-01-01

    The kinetics of charge transfer depend crucially on the dielectric reorganization of the medium. In enzymatic reactions that involve charge transfer, atomic dielectric response of the active site and of its surroundings determines the efficiency of the protein as a catalyst. We report direct spectroscopic measurements of the reorganization energy associated with the dielectric response in the active site of α-chymotrypsin. A chromophoric inhibitor of the enzyme is used as a spectroscopic probe. We find that water strongly affects the dielectric reorganization in the active site of the enzyme in solution. The reorganization energy of the protein matrix in the vicinity of the active site is similar to that of low-polarity solvents. Surprisingly, water exhibits an anomalously high dielectric response that cannot be described in terms of the dielectric continuum theory. As a result, sequestering the active site from the aqueous environment inside low-dielectric enzyme body dramatically reduces the dielectric reorganization. This reduction is particularly important for controlling the rate of enzymatic reactions. PMID:10681440

  11. Results of chemical analyses of soil, shale, and soil/shale extract from the Mancos Shale formation in the Gunnison Gorge National Conservation Area, southwestern Colorado, and at Hanksville, Utah

    USGS Publications Warehouse

    Tuttle, Michele L.W.; Fahy, Juli; Grauch, Richard I.; Ball, Bridget A.; Chong, Geneva W.; Elliott, John G.; Kosovich, John J.; Livo, Keith E.; Stillings, Lisa L.

    2007-01-01

    Results of chemical and some isotopic analyses of soil, shale, and water extracts collected from the surface, trenches, and pits in the Mancos Shale are presented in this report. Most data are for sites on the Gunnison Gorge National Conservation Area (GGNCA) in southwestern Colorado. For comparison, data from a few sites from the Mancos landscape near Hanksville, Utah, are included. Twelve trenches were dug on the GGNCA from which 258 samples for whole-rock (total) analyses and 187 samples for saturation paste extracts were collected. Sixteen of the extract samples were duplicated and subjected to a 1:5 water extraction for comparison. A regional soil survey across the Mancos landscape on the GGNCA generated 253 samples for whole-rock analyses and saturation paste extractions. Seventeen gypsum samples were collected on the GGNCA for sulfur and oxygen isotopic analysis. Sixteen samples were collected from shallow pits in the Mancos Shale near Hanksville, Utah.

  12. Methanopyrus kandleri topoisomerase V contains three distinct AP lyase active sites in addition to the topoisomerase active site

    PubMed Central

    Rajan, Rakhi; Osterman, Amy; Mondragón, Alfonso

    2016-01-01

    Topoisomerase V (Topo-V) is the only topoisomerase with both topoisomerase and DNA repair activities. The topoisomerase activity is conferred by a small alpha-helical domain, whereas the AP lyase activity is found in a region formed by 12 tandem helix-hairpin-helix ((HhH)2) domains. Although it was known that Topo-V has multiple repair sites, only one had been mapped. Here, we show that Topo-V has three AP lyase sites. The atomic structure and Small Angle X-ray Scattering studies of a 97 kDa fragment spanning the topoisomerase and 10 (HhH)2 domains reveal that the (HhH)2 domains extend away from the topoisomerase domain. A combination of biochemical and structural observations allow the mapping of the second repair site to the junction of the 9th and 10th (HhH)2 domains. The second site is structurally similar to the first one and to the sites found in other AP lyases. The 3rd AP lyase site is located in the 12th (HhH)2 domain. The results show that Topo-V is an unusual protein: it is the only known protein with more than one (HhH)2 domain, the only known topoisomerase with dual activities and is also unique by having three AP lyase repair sites in the same polypeptide. PMID:26908655

  13. Total Nitrogen Sources of the Three Gorges Reservoir — A Spatio-Temporal Approach

    PubMed Central

    Ren, Chunping; Wang, Lijing; Zheng, Binghui; Holbach, Andreas

    2015-01-01

    Understanding the spatial and temporal variation of nutrient concentrations, loads, and their distribution from upstream tributaries is important for the management of large lakes and reservoirs. The Three Gorges Dam was built on the Yangtze River in China, the world’s third longest river, and impounded the famous Three Gorges Reservoir (TGR). In this study, we analyzed total nitrogen (TN) concentrations and inflow data from 2003 till 2010 for the main upstream tributaries of the TGR that contribute about 82% of the TGR’s total inflow. We used time series analysis for seasonal decomposition of TN concentrations and used non-parametric statistical tests (Kruskal-Walli H, Mann-Whitney U) as well as base flow segmentation to analyze significant spatial and temporal patterns of TN pollution input into the TGR. Our results show that TN concentrations had significant spatial heterogeneity across the study area (Tuo River> Yangtze River> Wu River> Min River> Jialing River>Jinsha River). Furthermore, we derived apparent seasonal changes in three out of five upstream tributaries of the TGR rivers (Kruskal-Walli H ρ = 0.009, 0.030 and 0.029 for Tuo River, Jinsha River and Min River in sequence). TN pollution from non-point sources in the upstream tributaries accounted for 68.9% of the total TN input into the TGR. Non-point source pollution of TN revealed increasing trends for 4 out of five upstream tributaries of the TGR. Land use/cover and soil type were identified as the dominant driving factors for the spatial distribution of TN. Intensifying agriculture and increasing urbanization in the upstream catchments of the TGR were the main driving factors for non-point source pollution of TN increase from 2003 till 2010. Land use and land cover management as well as chemical fertilizer use restriction were needed to overcome the threats of increasing TN pollution. PMID:26510158

  14. Spatial bedrock erosion distribution in a natural gorge

    NASA Astrophysics Data System (ADS)

    Beer, A. R.; Turowski, J. M.; Kirchner, J. W.

    2015-12-01

    Quantitative analysis of morphological evolution both in terrestrial and planetary landscapes is of increasing interest in the geosciences. In mountainous regions, bedrock channel formation as a consequence of the interaction of uplift and erosion processes is fundamental for the entire surface evolution. Hence, the accurate description of bedrock channel development is important for landscape modelling. To verify existing concepts developed in the lab and to analyse how in situ channel erosion rates depend on the interrelations of discharge, sediment transport and topography, there is a need of highly resolved topographic field data. We analyse bedrock erosion over two years in a bedrock gorge downstream of the Gorner glacier above the town of Zermatt, Switzerland. At the study site, the Gornera stream cuts through a roche moutonnée in serpentine rock of 25m length, 5m width and 8m depth. We surveyed bedrock erosion rates using repeat terrestrial laser scanning (TLS) with an average point spacing of 5mm. Bedrock erosion rates in direction of the individual surface normals were studied directly on the scanned point clouds applying the M3C2 algorithm (Lague et al., 2013, ISPRS). The surveyed erosion patterns were compared to a simple stream erosivity visualisation obtained from painted bedrock sections at the study location. Spatially distributed erosion rates on bedrock surfaces based on millions of scan points allow deduction of millimeter-scale mean annual values of lateral erosion, incision and downstream erosion on protruding streambed surfaces. The erosion rate on a specific surface point is shown to depend on the position of this surface point in the channel's cross section, its height above the streambed and its spatial orientation to the streamflow. Abrasion by impacting bedload was likely the spatially dominant erosion process, as confirmed by the observed patterns along the painted bedrock sections. However, a single plucking event accounted for the half

  15. Nut Cracking Tools Used by Captive Chimpanzees (Pan troglodytes) and Their Comparison with Early Stone Age Percussive Artefacts from Olduvai Gorge.

    PubMed

    Arroyo, Adrián; Hirata, Satoshi; Matsuzawa, Tetsuro; de la Torre, Ignacio

    2016-01-01

    We present the results of a series of experiments at the Kumamoto Sanctuary in Japan, in which captive chimpanzees (Pan troglodytes) performed several nut cracking sessions using raw materials from Olduvai Gorge, Tanzania. We examined captive chimpanzee pounding tools using a combination of technological analysis, use-wear distribution, and micro-wear analysis. Our results show specific patterns of use-wear distribution across the active surfaces of pounding tools, which reveal some similarities with traces on archaeological percussive objects from the Early Stone Age, and are consistent with traces on other experimental pounding tools used by modern humans. The approach used in this study may help to stablish a framework with which to interpret archaeological assemblages and improve understanding of use-wear formation processes on pounding tools used by chimpanzees. This study represents the first direct comparison of chimpanzee pounding tools and archaeological material, and thus may contribute to a better understanding of hominin percussive activities.

  16. Nut Cracking Tools Used by Captive Chimpanzees (Pan troglodytes) and Their Comparison with Early Stone Age Percussive Artefacts from Olduvai Gorge

    PubMed Central

    Arroyo, Adrián; Hirata, Satoshi; Matsuzawa, Tetsuro; de la Torre, Ignacio

    2016-01-01

    We present the results of a series of experiments at the Kumamoto Sanctuary in Japan, in which captive chimpanzees (Pan troglodytes) performed several nut cracking sessions using raw materials from Olduvai Gorge, Tanzania. We examined captive chimpanzee pounding tools using a combination of technological analysis, use-wear distribution, and micro-wear analysis. Our results show specific patterns of use-wear distribution across the active surfaces of pounding tools, which reveal some similarities with traces on archaeological percussive objects from the Early Stone Age, and are consistent with traces on other experimental pounding tools used by modern humans. The approach used in this study may help to stablish a framework with which to interpret archaeological assemblages and improve understanding of use-wear formation processes on pounding tools used by chimpanzees. This study represents the first direct comparison of chimpanzee pounding tools and archaeological material, and thus may contribute to a better understanding of hominin percussive activities. PMID:27870877

  17. Use of Cusp Catastrophe for Risk Analysis of Navigational Environment: A Case Study of Three Gorges Reservoir Area

    PubMed Central

    Hao, Guozhu

    2016-01-01

    A water traffic system is a huge, nonlinear, complex system, and its stability is affected by various factors. Water traffic accidents can be considered to be a kind of mutation of a water traffic system caused by the coupling of multiple navigational environment factors. In this study, the catastrophe theory, principal component analysis (PCA), and multivariate statistics are integrated to establish a situation recognition model for a navigational environment with the aim of performing a quantitative analysis of the situation of this environment via the extraction and classification of its key influencing factors; in this model, the natural environment and traffic environment are considered to be two control variables. The Three Gorges Reservoir area of the Yangtze River is considered as an example, and six critical factors, i.e., the visibility, wind, current velocity, route intersection, channel dimension, and traffic flow, are classified into two principal components: the natural environment and traffic environment. These two components are assumed to have the greatest influence on the navigation risk. Then, the cusp catastrophe model is employed to identify the safety situation of the regional navigational environment in the Three Gorges Reservoir area. The simulation results indicate that the situation of the navigational environment of this area is gradually worsening from downstream to upstream. PMID:27391057

  18. Use of Cusp Catastrophe for Risk Analysis of Navigational Environment: A Case Study of Three Gorges Reservoir Area.

    PubMed

    Jiang, Dan; Hao, Guozhu; Huang, Liwen; Zhang, Dan

    2016-01-01

    A water traffic system is a huge, nonlinear, complex system, and its stability is affected by various factors. Water traffic accidents can be considered to be a kind of mutation of a water traffic system caused by the coupling of multiple navigational environment factors. In this study, the catastrophe theory, principal component analysis (PCA), and multivariate statistics are integrated to establish a situation recognition model for a navigational environment with the aim of performing a quantitative analysis of the situation of this environment via the extraction and classification of its key influencing factors; in this model, the natural environment and traffic environment are considered to be two control variables. The Three Gorges Reservoir area of the Yangtze River is considered as an example, and six critical factors, i.e., the visibility, wind, current velocity, route intersection, channel dimension, and traffic flow, are classified into two principal components: the natural environment and traffic environment. These two components are assumed to have the greatest influence on the navigation risk. Then, the cusp catastrophe model is employed to identify the safety situation of the regional navigational environment in the Three Gorges Reservoir area. The simulation results indicate that the situation of the navigational environment of this area is gradually worsening from downstream to upstream.

  19. Bed II Sequence Stratigraphic context of EF-HR and HWK EE archaeological sites, and the Oldowan/Acheulean succession at Olduvai Gorge, Tanzania.

    PubMed

    Stanistreet, Ian G; McHenry, Lindsay J; Stollhofen, Harald; de la Torre, Ignacio

    2018-04-20

    Archaeological excavations at EF-HR and HWK EE allow reassessment of Bed II stratigraphy within the Junction Area and eastern Olduvai Gorge. Application of Sequence Stratigraphic methods provides a time-stratigraphic framework enabling correlation of sedimentary units across facies boundaries, applicable even in those areas where conventional timelines, such as tephrostratigraphic markers, are absent, eroded, or reworked. Sequence Stratigraphically, Bed II subdivides into five major Sequences 1 to 5, all floored by major disconformities that incise deeply into the underlying succession, proving that simple "layer cake" stratigraphy is inappropriate. Previous establishment of the Lemuta Member has invalidated the use of Tuff IIA as the boundary between Lower and Middle Bed II, now redefined at the disconformity between Sequences 2 and 3, a lithostratigraphic contact underlying the succession containing the Lower, Middle, and Upper Augitic Sandstones. HWK EE site records Oldowan technology in the Lower Augitic Sandstone at the base of Sequence 3, within Middle Bed II. We suggest placement of recently reported Acheulean levels at FLK W within the Middle Augitic Sandstone, thus emphasizing that handaxes are yet to be found in earlier stratigraphic units of the Olduvai sequence. This would place a boundary between the Oldowan and Acheulean technologies at Olduvai in the Tuff IIB zone or earliest Middle Augitic Sandstone. A major disconformity between Sequences 3 and 4 at and near EF-HR cuts through the level of Tuff IIC, placing the main Acheulean EF-HR assemblage at the base of Sequence 4, within Upper rather than Middle Bed II. Sequence stratigraphic methods also yield a more highly resolved Bed II stratigraphic framework. Backwall and sidewall surveying of archaeological trenches at EF-HR and HWK EE permits definition of "Lake-parasequences" nested within the major Sequences that record downcutting of disconformities associated with lake regression, then

  20. The Three Gorges Dam Affects Regional Precipitation

    NASA Technical Reports Server (NTRS)

    Wu, Liguang; Zhang, Qiang; Jiang, Zhihong

    2006-01-01

    Issues regarding building large-scale dams as a solution to power generation and flood control problems have been widely discussed by both natural and social scientists from various disciplines, as well as the policy-makers and public. Since the Chinese government officially approved the Three Gorges Dam (TGD) projects, this largest hydroelectric project in the world has drawn a lot of debates ranging from its social and economic to climatic impacts. The TGD has been partially in use since June 2003. The impact of the TGD is examined through analysis of the National Aeronautics and Space Administration (NASA) Tropical Rainfall Measuring Mission (TRMM) rainfall rate and Moderate Resolution Imaging Spectroradiometer (MODIS) land surface temperature and high-resolution simulation using the Pennsylvania State University-National Center for Atmospheric Research (PSU-NCAR) fifth-generation Mesoscale Model (MM5). The independent satellite data sets and numerical simulation clearly indicate that the land use change associated with the TGD construction has increased the precipitation in the region between Daba and Qinling mountains and reduced the precipitation in the vicinity of the TGD after the TGD water level abruptly rose from 66 to 135 m in June 2003. This study suggests that the climatic effect of the TGD is on the regional scale (approx.100 km) rather than on the local scale (approx.10 km) as projected in previous studies.

  1. Sedimentary record and luminescence chronology of palaeoflood events along the Gold Gorge of the upper Hanjiang River, middle Yangtze River basin, China

    NASA Astrophysics Data System (ADS)

    Guo, Yongqiang; Huang, Chun Chang; Zhou, Yali; Pang, Jiangli; Zha, Xiaochun; Fan, Longjiang; Mao, Peini

    2018-05-01

    Palaeoflood slackwater deposits (SWDs) along the river banks have important implications for the reconstruction of the past hydro-climatic events. Two palaeoflood SWD beds were identified in the Holocene loess-soil sequences on the cliff river banks along the Gold Gorge of the upper Hanjiang River by field investigation and laboratory analysis. They have recorded two palaeoflood events which were dated by optically stimulated luminescence to 3.2-2.8 ka and 2.1-1.8 ka, respectively. The reliability of the ages obtained for the two events are further confirmed by the presence of archaeological remains and good regional pedostratigraphic correlation. The peak discharges of two palaeoflood events at the studied sites were estimated to be 16,560-17,930 m3/s. A correlation with the palaeoflood events identified in the other reaches shows that great floods occurred frequently during the episodes of 3200-2800 and 2000-1700 a BP along the upper Hanjiang River valley during the last 4000 years. These phases of palaeoflood events in central China are well correlated with the climatic variability identified by δ18O record in the stalagmites from the middle Yangtze River Basin and show apparent global linkages. Palaeoflood studies in a watershed scale also imply that strengthened human activities during the Shang dynasty (BCE 1600-1100) and Han dynasty (BCE206-CE265) may have caused accelerated soil erosion along the upper Hanjiang River valley.

  2. Activation of phenylalanine hydroxylase by phenylalanine does not require binding in the active site.

    PubMed

    Roberts, Kenneth M; Khan, Crystal A; Hinck, Cynthia S; Fitzpatrick, Paul F

    2014-12-16

    Phenylalanine hydroxylase (PheH), a liver enzyme that catalyzes the hydroxylation of excess phenylalanine in the diet to tyrosine, is activated by phenylalanine. The lack of activity at low levels of phenylalanine has been attributed to the N-terminus of the protein's regulatory domain acting as an inhibitory peptide by blocking substrate access to the active site. The location of the site at which phenylalanine binds to activate the enzyme is unknown, and both the active site in the catalytic domain and a separate site in the N-terminal regulatory domain have been proposed. Binding of catecholamines to the active-site iron was used to probe the accessibility of the active site. Removal of the regulatory domain increases the rate constants for association of several catecholamines with the wild-type enzyme by ∼2-fold. Binding of phenylalanine in the active site is effectively abolished by mutating the active-site residue Arg270 to lysine. The k(cat)/K(phe) value is down 10⁴ for the mutant enzyme, and the K(m) value for phenylalanine for the mutant enzyme is >0.5 M. Incubation of the R270K enzyme with phenylalanine also results in a 2-fold increase in the rate constants for catecholamine binding. The change in the tryptophan fluorescence emission spectrum seen in the wild-type enzyme upon activation by phenylalanine is also seen with the R270K mutant enzyme in the presence of phenylalanine. Both results establish that activation of PheH by phenylalanine does not require binding of the amino acid in the active site. This is consistent with a separate allosteric site, likely in the regulatory domain.

  3. Activation of Phenylalanine Hydroxylase by Phenylalanine Does Not Require Binding in the Active Site

    PubMed Central

    2015-01-01

    Phenylalanine hydroxylase (PheH), a liver enzyme that catalyzes the hydroxylation of excess phenylalanine in the diet to tyrosine, is activated by phenylalanine. The lack of activity at low levels of phenylalanine has been attributed to the N-terminus of the protein’s regulatory domain acting as an inhibitory peptide by blocking substrate access to the active site. The location of the site at which phenylalanine binds to activate the enzyme is unknown, and both the active site in the catalytic domain and a separate site in the N-terminal regulatory domain have been proposed. Binding of catecholamines to the active-site iron was used to probe the accessibility of the active site. Removal of the regulatory domain increases the rate constants for association of several catecholamines with the wild-type enzyme by ∼2-fold. Binding of phenylalanine in the active site is effectively abolished by mutating the active-site residue Arg270 to lysine. The kcat/Kphe value is down 104 for the mutant enzyme, and the Km value for phenylalanine for the mutant enzyme is >0.5 M. Incubation of the R270K enzyme with phenylalanine also results in a 2-fold increase in the rate constants for catecholamine binding. The change in the tryptophan fluorescence emission spectrum seen in the wild-type enzyme upon activation by phenylalanine is also seen with the R270K mutant enzyme in the presence of phenylalanine. Both results establish that activation of PheH by phenylalanine does not require binding of the amino acid in the active site. This is consistent with a separate allosteric site, likely in the regulatory domain. PMID:25453233

  4. Non-competitive inhibition by active site binders.

    PubMed

    Blat, Yuval

    2010-06-01

    Classical enzymology has been used for generations to understand the interactions of inhibitors with their enzyme targets. Enzymology tools enabled prediction of the biological impact of inhibitors as well as the development of novel, more potent, ones. Experiments designed to examine the competition between the tested inhibitor and the enzyme substrate(s) are the tool of choice to identify inhibitors that bind in the active site. Competition between an inhibitor and a substrate is considered a strong evidence for binding of the inhibitor in the active site, while the lack of competition suggests binding to an alternative site. Nevertheless, exceptions to this notion do exist. Active site-binding inhibitors can display non-competitive inhibition patterns. This unusual behavior has been observed with enzymes utilizing an exosite for substrate binding, isomechanism enzymes, enzymes with multiple substrates and/or products and two-step binding inhibitors. In many of these cases, the mechanisms underlying the lack of competition between the substrate and the inhibitor are well understood. Tools like alternative substrates, testing the enzyme reaction in the reverse direction and monitoring inhibition time dependence can be applied to enable distinction between 'badly behaving' active site binders and true exosite inhibitors.

  5. Tectonic evolution of the central Brooks Range mountain front: Evidence from the Atigun Gorge region

    USGS Publications Warehouse

    Mull, C.G.; Glenn, R.K.; Adams, K.E.

    1997-01-01

    Atigun Gorge, at the northern front of the eastern Endicott Mountains, contains well-exposed rocks of the upper part of the Endicott Mountains allochthon and rocks of the structurally higher Picnic Creek or Ipnavik River allochthon. These allochthons contain rocks as young as Early Cretaceous (Valanginian) and are separated by a nearly vertical fault zone that contains exotic blocks of Triassic and Jurassic chert and silicified mudstone. Siliceous rocks of this type are not present in the Endicott Mountains allochthon but are characteristic of the Picnic Creek, Ipnavik River, and some of the other allochthons that structurally overlie the Endicott Mountains allochthon in the central and western Brooks Range. These exotic blocks, therefore indicate that structurally higher rocks of either the Picnic Creek or Ipnavik River allochthon were emplaced during the Early Cretaceous and are preserved along the northern flank of the eastern Endicott Mountains. The deformed thickness of this higher allochthon in the subsurface north of the mountains is unknown but probably exceeds 2 kilometers. Similar relations are mapped east of Atigun Gorge in an area of structural transition from the eastern Endicott Mountains into the northern Philip Smith Mountains, which are formed by the parautochthonous North Slope stratigraphic assemblage. The allochthonous rocks at the mountain front are regionally unconformably overlain by proximal Lower Cretaceous (Albian) foredeep conglomerate at the southern flank of the Colville basin, but at Atigun Gorge, the base of these deposits is interpreted as a possible back thrust at a triangle zone. Conglomerate clasts in the foredeep deposits are dominantly chert, mafic igneous rock, and other lithologies characteristic of the Picnic Creek and Ipnavik River allochthons and scattered clasts from the Endicott Mountains allochthon. The conglomerates show that the chert-rich allochthonous rocks and the Endicott Mountains allochthon were emplaced in the

  6. Cataclysms and controversy -- aspects of the geomorphology of the Columbia River Gorge

    USGS Publications Warehouse

    O'Connor, Jim; Burns, Scott; Madin, Ian; Dorsey, Rebecca

    2009-01-01

    Landslides and floods of lava and water tremendously affected the Columbia River during its long history of transecting the Cascade Volcanic Arc. This field trip touches on aspects of the resulting geology of the scenic Columbia River Gorge, including the river-blocking Bonneville landslide of ~550 years ago and the great late- Pleistocene Missoula floods. Not only did these events create great landscapes, but they inspired great geologists. Mid-nineteenth century observations of the Columbia River and Pacific Northwest by James Dwight Dana and John Strong Newberry helped germinate the “school of fluvial” erosion later expanded upon by the southwestern United States topographic and geologic surveys. Later work on features related to the Missoula floods framed the career of J Harlen Bretz in one of the great geologic controversies of the twentieth century.

  7. Demonstration of three gorges archaeological relics based on 3D-visualization technology

    NASA Astrophysics Data System (ADS)

    Xu, Wenli

    2015-12-01

    This paper mainly focuses on the digital demonstration of three gorges archeological relics to exhibit the achievements of the protective measures. A novel and effective method based on 3D-visualization technology, which includes large-scaled landscape reconstruction, virtual studio, and virtual panoramic roaming, etc, is proposed to create a digitized interactive demonstration system. The method contains three stages: pre-processing, 3D modeling and integration. Firstly, abundant archaeological information is classified according to its history and geographical information. Secondly, build up a 3D-model library with the technology of digital images processing and 3D modeling. Thirdly, use virtual reality technology to display the archaeological scenes and cultural relics vividly and realistically. The present work promotes the application of virtual reality to digital projects and enriches the content of digital archaeology.

  8. Safety Oversight of Decommissioning Activities at DOE Nuclear Sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zull, Lawrence M.; Yeniscavich, William

    2008-01-15

    The Defense Nuclear Facilities Safety Board (Board) is an independent federal agency established by Congress in 1988 to provide nuclear safety oversight of activities at U.S. Department of Energy (DOE) defense nuclear facilities. The activities under the Board's jurisdiction include the design, construction, startup, operation, and decommissioning of defense nuclear facilities at DOE sites. This paper reviews the Board's safety oversight of decommissioning activities at DOE sites, identifies the safety problems observed, and discusses Board initiatives to improve the safety of decommissioning activities at DOE sites. The decommissioning of former defense nuclear facilities has reduced the risk of radioactive materialmore » contamination and exposure to the public and site workers. In general, efforts to perform decommissioning work at DOE defense nuclear sites have been successful, and contractors performing decommissioning work have a good safety record. Decommissioning activities have recently been completed at sites identified for closure, including the Rocky Flats Environmental Technology Site, the Fernald Closure Project, and the Miamisburg Closure Project (the Mound site). The Rocky Flats and Fernald sites, which produced plutonium parts and uranium materials for defense needs (respectively), have been turned into wildlife refuges. The Mound site, which performed R and D activities on nuclear materials, has been converted into an industrial and technology park called the Mound Advanced Technology Center. The DOE Office of Legacy Management is responsible for the long term stewardship of these former EM sites. The Board has reviewed many decommissioning activities, and noted that there are valuable lessons learned that can benefit both DOE and the contractor. As part of its ongoing safety oversight responsibilities, the Board and its staff will continue to review the safety of DOE and contractor decommissioning activities at DOE defense nuclear sites.« less

  9. Prevalence of pansteatitis in African sharptooth catfish, Clarias gariepinus (Burchell), in the Kruger National Park, South Africa.

    PubMed

    Huchzermeyer, K David A

    2012-11-09

    Pansteatitis was confirmed in sharptooth catfish, Clarias gariepinus (Burchell), from three main locations within the Kruger National Park (KNP); the Olifants River Gorge, Engelhard Dam on the Letaba River and from the Sabie River in the Sabiepoort. An increasing prevalence of pansteatitis was observed in catfish during repeated samplings from the Olifants Gorge from 2009 to 2011 and co-existence of old and recent lesions indicated on-going incitement of pansteatitis. Only a low prevalence of pansteatitis was observed in catfish sampled from the Olifants River upstream of the Gorge in the KNP and no pansteatitis was observed in catfish sampled from a rain-filled dam not connected to the Olifants River. Common to both the Olifants Gorge and the Sabiepoort is the damming of the rivers in Mozambique to form lakes Massingir and Corumana respectively. Anthropogenic activities resulting in potential pollution of the rivers differ greatly between these two catchments, providing argument against a primary pollution-related aetiology of the pansteatitis found at these two sites. Compared with other sites, analysis of stomach contents of catfish from the Olifants Gorge and the Sabiepoort strongly suggested that consumption of a predominantly fish diet was associated with the development of pansteatitis in these fish. In a farmed population of catfish used as positive control, development of pansteatitis could be ascribed to consumption of rancid fish waste from a trout slaughterhouse. In the Olifants Gorge, alien invasive silver carp, Hypophthalmychthys molitrix (Valenciennes), seasonally migrate upstream out of Lake Massingir to spawn. This schooling species is an obligate phytoplankton feeder with consequent high levels of adipose tissue n-3 polyunsaturated fatty acids. In the Olifants Gorge, at least, this may explain seasonal exposure to levels of polyunsaturated fats in the diets of catfish and crocodiles to which these animals are not adapted. The possible roles of

  10. Reconstructing the 1935 Columbia River Gorge: A Topographic and Orthophoto Experiment

    NASA Astrophysics Data System (ADS)

    Fonstad, M. A.; Major, J. H.; O'Connor, J. E.; Dietrich, J. T.

    2017-12-01

    The last decade has seen a revolution in the mapping of rivers and near-river environments. Much of this has been associated with a new type of photogrammetry: structure from motion (SfM) and multi-view stereo techniques. Through SfM, 3D surfaces are reconstructed from nonstructured image groups with poorly calibrated cameras whose locations need not be known. Modern SfM imaging is greatly improved by careful flight planning, well-planned ground control or high-precision direct georeferencing, and well-understood camera optics. The ease of SfM, however, begs the question: how well does it work on archival photos taken without the foreknowledge of SfM techniques? In 1935, the Army Corps of Engineers took over 800 vertical aerial photos for a 160-km-long stretch of the Columbia River Gorge and adjacent areas in Oregon and Washington. These photos pre-date completion of three hydroelectric dams and reservoirs in this reach, and thus provide rich information on the historic pre-dam riverine, geologic, and cultural environments. These photos have little to no metadata associated with them, such as camera calibration reports, so traditional photogrammetry techniques are exceeding difficult to apply. Instead, we apply SfM to these archival photos, and test the resulting digital elevation model (DEM) against lidar data for features inferred to be unchanged in the past 80 years. Few, if any, of the quality controls recommended for SfM are available for these 1935 photos; they are scanned paper positives with little overlap taken with an unknown optical system in high altitude flight paths. Nevertheless, in almost all areas, the SfM analysis produced a high quality orthophoto of the gorge with low horizontal errors - most better than a few meters. The DEM created looks highly realistic, and in many areas has a vertical error of a few meters. However, the vertical errors are spatially inconsistent, with some wildly large, likely because of the many poorly constrained links in

  11. Towards a better knowledge of flash flood forecasting at the Three Gorges Region: Progress over the past decade and challenges ahead

    NASA Astrophysics Data System (ADS)

    Li, Zhe; Yang, Dawen; Yang, Hanbo; Wu, Tianjiao; Xu, Jijun; Gao, Bing; Xu, Tao

    2015-04-01

    The study area, the Three Gorges Region (TGR), plays a critical role in predicting the floods drained into the Three Gorges Reservoir, as reported local floods often exceed 10000m3/s during rainstorm events and trigger fast as well as significant impacts on the Three Gorges Reservoir's regulation. Meanwhile, it is one of typical mountainous areas in China, which is located in the transition zone between two monsoon systems: the East Asian monsoon and the South Asian (Indian) monsoon. This climatic feature, combined with local irregular terrains, has shaped complicated rainfall-runoff regimes in this focal region. However, due to the lack of high-resolution hydrometeorological data and physically-based hydrologic modeling framework, there was little knowledge about rainfall variability and flood pattern in this historically ungauged region, which posed great uncertainties to flash flood forecasting in the past. The present study summarize latest progresses of regional flash floods monitoring and prediction, including installation of a ground-based Hydrometeorological Observation Network (TGR-HMON), application of a regional geomorphology-based hydrological model (TGR-GBHM), development of an integrated forecasting and modeling system (TGR-INFORMS), and evaluation of quantitative precipitation estimations (QPE) and quantitative precipitation forecasting (QPF) products in TGR flash flood forecasting. With these continuing efforts to improve the forecasting performance of flash floods in TGR, we have addressed several critical issues: (1) Current observation network is still insufficient to capture localized rainstorms, and weather radar provides valuable information to forecast flash floods induced by localized rainstorms, although current radar QPE products can be improved substantially in future; (2) Long-term evaluation shows that the geomorphology-based distributed hydrologic model (GBHM) is able to simulate flash flooding processes reasonably, while model

  12. Research on the Placement of the Ecological Shelter Zone in the Three Gorges Reservoir Area, China

    NASA Astrophysics Data System (ADS)

    Shan, N.; Ruan, X.

    2011-12-01

    The Three Gorges Dam is built on the middle reaches of Yangtze River (Changjiang) in south-central China, which is the world's third longest river. The Three Gorges Reservoir Region (TGRR), including the entire inundated area and 19 administrative units (counties and cities) on both sides of the river, is regarded as an environmentally sensitive area. The total area of the TGRR is approximately 58000 km2. As the Three Gorges Dam fully operated, for the flood control, the water level should be kept in the range between 145 m and 175 m and the reservoir surface water area(over 1080 km2)at a water level of 175 m, with a length of 600 km. Many of cities, villages and farms have been submerged. Moreover, as a result of reservoir operation, the water-level alternation of the reservoir is opposite to the nature, which is low water level (145m) in summer and high water level (175m) in winter. The Hydro-Fluctuation Belt, with a height of 30m, will become a new pollution source due to the riparian being flooded and the submerged areas may still contain trace amounts of toxic or radioactive materials. The environmental impacts associated with large scale reservoir area often have significant negative impacts on the environment. It affects forest cover, species in the area, some endangered, water quality, increase the likelihood of earthquakes and mudslides in the area. To solve these problems, it is necessarily to construct the Ecological Shelter Zone (ESZ) along with the edge of the reservoir area. The function of the ESZ is similar to the riparian zone in reducing flood damage, improving water quality, decreasing the levels of the nonpoint source pollution load and soil erosion and rebuilding the migration routes of plant and wildlife. However, the research of the ESZ is mainly focused on rivers at field scale by now, lack of research method on reservoir at the watershed scale. As the special nature of the Three Gorges Reservoir, the construction of the ESZ in the TGRA is

  13. Species-level phylogeographical history of Myricaria plants in the mountain ranges of western China and the origin of M. laxiflora in the Three Gorges mountain region.

    PubMed

    Liu, Yifei; Wang, Yong; Huang, Hongwen

    2009-06-01

    Myricaria species in China occur mostly in the major high-altitude mountain areas in and around the Qinghai-Tibetan Plateau. The one major exception to this is M. laxiflora which is restricted to the Three Gorges mountain region. In this study, we investigate species-level phylogeographical patterns of Myricaria species in western China and the origin of M. laxiflora. The results show that most chloroplast haplotypes are species-specific, except for one haplotype which is shared by three widespread species. Higher haplotype diversity within the Qinghai-Tibetan Plateau region supports the hypothesis that the Himalayas are the centre of origin for Myricaria. The phylogeny of Myricaria was geographically structured, and an estimated Bayesian chronology suggested the main divergence events occurred during the Late Pliocene and Early Pleistocene (approximately 1.46-2.30 million years ago). The overall phylogeographical pattern was characterized by vicariance events and regional demographical expansion, reflecting a major influence of geological and climatic events on the evolution of Myricaria species. Our data suggest that M. laxiflora has an ancient origin, but has experienced recent population expansion through the Three Gorges Valley. The origin of M. laxiflora was estimated to be during the Early Pleistocene but its demographical expansion was more recent at about 0.015 million years ago. This highlights the unique phylogeographical history of the Three Gorges mountain region, and the deep imprint of the watercourse connections of the Yangtze River Valley on the phylogeographical structure of the species in this region.

  14. Ionizable Side Chains at Catalytic Active Sites of Enzymes

    PubMed Central

    Jimenez-Morales, David; Liang, Jie

    2012-01-01

    Catalytic active sites of enzymes of known structure can be well defined by a modern program of computational geometry. The CASTp program was used to define and measure the volume of the catalytic active sites of 573 enzymes in the Catalytic Site Atlas database. The active sites are identified as catalytic because the amino acids they contain are known to participate in the chemical reaction catalyzed by the enzyme. Acid and base side chains are reliable markers of catalytic active sites. The catalytic active sites have 4 acid and 5 base side chains, in an average volume of 1072 Å3. The number density of acid side chains is 8.3 M (in chemical units); the number density of basic side chains is 10.6 M. The catalytic active site of these enzymes is an unusual electrostatic and steric environment in which side chains and reactants are crowded together in a mixture more like an ionic liquid than an ideal infinitely dilute solution. The electrostatics and crowding of reactants and side chains seems likely to be important for catalytic function. In three types of analogous ion channels, simulation of crowded charges accounts for the main properties of selectivity measured in a wide range of solutions and concentrations. It seems wise to use mathematics designed to study interacting complex fluids when making models of the catalytic active sites of enzymes. PMID:22484856

  15. Water exposure assessment of aryl hydrocarbon receptor agonists in Three Gorges Reservoir, China using SPMD-based virtual organisms.

    PubMed

    Wang, Jingxian; Bernhöft, Silke; Pfister, Gerd; Schramm, Karl-Werner

    2014-10-15

    SPMD-based virtual organisms (VOs) were deployed at five to eight sites in the Three Gorges Reservoir (TGR), China for five periods in 2008, 2009 and 2011. The water exposure of aryl hydrocarbon receptor (AhR) agonists was assessed by the VOs. The chosen bioassay response for the extracts of the VOs, the induction of 7-ethoxyresorufin-O-deethylase (EROD) was assayed using a rat hepatoma cell line (H4IIE). The results show that the extracts from the VOs could induce AhR activity significantly, whereas the chemically derived 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalent (TEQcal) accounted for <11% of the observed AhR responses (TEQbio). Unidentified AhR-active compounds represented a greater proportion of the TCDD equivalent in VOs from TGR. High TEQbio value in diluted extract and low TEQbio in concentrated extract of the same sample was observed suggesting potential non-additive effects in the mixture. The levels of AhR agonists in VOs from upstream TGR were in general higher than those from downstream reservoir, indicating urbanization effect on AhR agonist pollution. The temporal variation showed that levels of AhR agonists in 2009 and 2011 were higher than those in 2008, and the potential non-additive effects in the area close to the dam were also obviously higher in 2009 and 2011 than in 2008, indicating big changes in the composition of pollutants in the area after water level reached a maximum of 175 m. Although the aqueous concentration of AhR agonists of 0.8-4.8 pg TCDDL(-1) in TGR was not alarming, the tendency of accumulating high concentration of AhR agonists in VO lipid and existence of possible synergism or antagonism in the water may exhibit a potential hazard to local biota being exposed to AhR agonists. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Discriminative structural approaches for enzyme active-site prediction.

    PubMed

    Kato, Tsuyoshi; Nagano, Nozomi

    2011-02-15

    Predicting enzyme active-sites in proteins is an important issue not only for protein sciences but also for a variety of practical applications such as drug design. Because enzyme reaction mechanisms are based on the local structures of enzyme active-sites, various template-based methods that compare local structures in proteins have been developed to date. In comparing such local sites, a simple measurement, RMSD, has been used so far. This paper introduces new machine learning algorithms that refine the similarity/deviation for comparison of local structures. The similarity/deviation is applied to two types of applications, single template analysis and multiple template analysis. In the single template analysis, a single template is used as a query to search proteins for active sites, whereas a protein structure is examined as a query to discover the possible active-sites using a set of templates in the multiple template analysis. This paper experimentally illustrates that the machine learning algorithms effectively improve the similarity/deviation measurements for both the analyses.

  17. Spatiotemporal distribution and risk assessment of organotins in the surface water of the Three Gorges Reservoir Region, China.

    PubMed

    Gao, Jun-Min; Wu, Lei; Chen, You-Peng; Zhou, Bin; Guo, Jin-Song; Zhang, Ke; Ouyang, Wen-Juan

    2017-03-01

    The water quality security of the Three Gorges Reservoir during different operating periods has been a subject of recent concern. This study is the first to report the spatiotemporal variability of organotins (OTs) in surface water under dynamic water level conditions in the Three Gorges Reservoir Region (TGRR). TGRR surface water was collected during three monitoring campaigns to analyze butyltins (BTs) and phenyltins (PTs) using a gas chromatography-mass spectrometry system. Our results showed that TGRR surface water was polluted by BTs and PTs, with mono-OTs being the dominant species. A wide range of BTs and PTs concentrations was observed across the study area, but tributyltin (TBT) displayed extensive spatial distribution, and the highest concentrations consistently occurred in the downstream region of the TGRR study area, with a maximum of 393.35 ng Sn/L in Zigui (S27). The total OTs contamination level decreased over time. The diphenyltin concentration exhibited significant seasonal variation, while other OTs showed seasonal changes only during two monitoring campaigns, with the exception of dibutyltin. An ecological risk assessment indicated that both TBT and triphenyltin posed risks to aquatic organisms in TGRR surface water. We urgently recommend continuous monitoring and further measures to prevent and control OTs pollution in the TGRR. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. [Effects of Three Gorges Reservoir impoundment on the wetland ecosystem service value of Dongting Lake, South-central China].

    PubMed

    Li, Jing-Bao; Dai, Yong; Yin, Ri-Xin; Yang, Yan; Li, Yu-dan; Wang, Ke-ying

    2013-03-01

    Based on the field investigation and measurement, and by using the monetary method, this paper estimated the wetland ecosystem service value of Dongting Lake before and after the impoundment of Three Gorges Reservoir (in 1996 and 2010, respectively). After the impoundment, the total ecosystem service value increased from 156.69x10(8) yuan in 1996 to 177.11x10(8) yuan in 2010. The main services value in 1996 was in the order of flood storage and regulation > water storage and supply > air regulation > scientific research and education, while that in 2010 was leisure tourism > shipping transportation > air regulation > water storage and supply. In the total service value of the wetland ecosystem, the direct value associated with water decreased from 110. 85x10(8) in 1996 to 27.47x10(8) in 2010, with a decrement of 75.2%. Though the proportion of the direct value in the production and supply of material products had somewhat increase, the indirect value in ecological environment regulation and maintenance and in culture and society still maintained at about 80% of the total value. In addition to climate factors, the impoundment of Three Gorges Reservoir and the reduction of water and sediment from Yangtze River to the Lake were the crucial reasons leading to the changes of the wetland ecosystem service value of Dongting Lake.

  19. Three-Gorge Reservoir: A 'Controlled Experiment' for Calibration/Validation of Time-Variable Gravity Signals Detected from Space

    NASA Technical Reports Server (NTRS)

    Chao, Benjamin F.; Boy, J. P.

    2003-01-01

    With the advances of measurements, modern space geodesy has become a new type of remote sensing for the Earth dynamics, especially for mass transports in the geophysical fluids on large spatial scales. A case in point is the space gravity mission GRACE (Gravity Recovery And Climate Experiment) which has been in orbit collecting gravity data since early 2002. The data promise to be able to detect changes of water mass equivalent to sub-cm thickness on spatial scale of several hundred km every month or so. China s Three-Gorge Reservoir has already started the process of water impoundment in phases. By 2009,40 km3 of water will be stored behind one of the world s highest dams and spanning a section of middle Yangtze River about 600 km in length. For the GRACE observations, the Three-Gorge Reservoir would represent a geophysical controlled experiment , one that offers a unique opportunity to do detailed geophysical studies. -- Assuming a complete documentation of the water level and history of the water impoundment process and aided with a continual monitoring of the lithospheric loading response (such as in area gravity and deformation), one has at hand basically a classical forwardinverse modeling problem of surface loading, where the input and certain output are known. The invisible portion of the impounded water, i.e. underground storage, poses either added values as an observable or a complication as an unknown to be modeled. Wang (2000) has studied the possible loading effects on a local scale; we here aim for larger spatial scales upwards from several hundred km, with emphasis on the time-variable gravity signals that can be detected by GRACE and follow-on missions. Results using the Green s function approach on the PREM elastic Earth model indicate the geoid height variations reaching several millimeters on wavelengths of about a thousand kilometers. The corresponding vertical deformations have amplitude of a few centimeters. In terms of long

  20. Ionizable side chains at catalytic active sites of enzymes.

    PubMed

    Jimenez-Morales, David; Liang, Jie; Eisenberg, Bob

    2012-05-01

    Catalytic active sites of enzymes of known structure can be well defined by a modern program of computational geometry. The CASTp program was used to define and measure the volume of the catalytic active sites of 573 enzymes in the Catalytic Site Atlas database. The active sites are identified as catalytic because the amino acids they contain are known to participate in the chemical reaction catalyzed by the enzyme. Acid and base side chains are reliable markers of catalytic active sites. The catalytic active sites have 4 acid and 5 base side chains, in an average volume of 1,072 Å(3). The number density of acid side chains is 8.3 M (in chemical units); the number density of basic side chains is 10.6 M. The catalytic active site of these enzymes is an unusual electrostatic and steric environment in which side chains and reactants are crowded together in a mixture more like an ionic liquid than an ideal infinitely dilute solution. The electrostatics and crowding of reactants and side chains seems likely to be important for catalytic function. In three types of analogous ion channels, simulation of crowded charges accounts for the main properties of selectivity measured in a wide range of solutions and concentrations. It seems wise to use mathematics designed to study interacting complex fluids when making models of the catalytic active sites of enzymes.

  1. Building Connections among Lands, People and Communities: A Case Study of Benefits-Based Management Plan Development for the Gunnison Gorge National Conservation Area

    Treesearch

    Richard C. Knopf; Kathleen L. Andereck; Karen Tucker; Bill Bottomly; Randy J. Virden

    2004-01-01

    Purpose of Study This paper demonstrates how a Benefits-Based Management paradigm has been useful in guiding management plan development for an internationally significant natural resource – the Gunnison Gorge National Conservation Area (GGNCA) in Colorado. Through a program of survey research, a database on benefits desired by various stakeholder groups was created....

  2. NDVI indicated long-term interannual changes in vegetation activities and their responses to climatic and anthropogenic factors in the Three Gorges Reservoir Region, China.

    PubMed

    Wen, Zhaofei; Wu, Shengjun; Chen, Jilong; Lü, Mingquan

    2017-01-01

    Natural and social environmental changes in the China's Three Gorges Reservoir Region (TGRR) have received worldwide attention. Identifying interannual changes in vegetation activities in the TGRR is an important task for assessing the impact these changes have on the local ecosystem. We used long-term (1982-2011) satellite-derived Normalized Difference Vegetation Index (NDVI) datasets and climatic and anthropogenic factors to analyze the spatiotemporal patterns of vegetation activities in the TGRR, as well as their links to changes in temperature (TEM), precipitation (PRE), downward radiation (RAD), and anthropogenic activities. At the whole TGRR regional scale, a statistically significant overall uptrend in NDVI variations was observed in 1982-2011. More specifically, there were two distinct periods with different trends split by a breakpoint in 1991: NDVI first sharply increased prior to 1991, and then showed a relatively weak rate of increase after 1991. At the pixel scale, most parts of the TGRR experienced increasing NDVI before the 1990s but different trend change types after the 1990s: trends were positive in forests in the northeastern parts, but negative in farmland in southwest parts of the TGRR. The TEM warming trend was the main climate-related driver of uptrending NDVI variations pre-1990s, and decreasing PRE was the main climate factor (42%) influencing the mid-western farmland areas' NDVI variations post-1990s. We also found that anthropogenic factors such as population density, man-made ecological restoration, and urbanization have notable impacts on the TGRR's NDVI variations. For example, large overall trend slopes in NDVI were more likely to appear in TGRR regions with large fractions of ecological restoration within the last two decades. The findings of this study may help to build a better understanding of the mechanics of NDVI variations in the periods before and during TGDP construction for ongoing ecosystem monitoring and assessment in the

  3. An in-silico insight into the substrate binding characteristics of the active site of amorpha-4, 11-diene synthase, a key enzyme in artemisinin biosynthesis.

    PubMed

    Eslami, Habib; Mohtashami, Seyed Kaveh; Basmanj, Maryam Taghavi; Rahati, Maryam; Rahimi, Hamzeh

    2017-07-01

    The enzyme amorphadiene synthase (ADS) conducts the first committed step in the biosynthetic conversion of the substrate farnesyl pyrophosphate (FPP) to artemisinin, which is a highly effective natural product against multidrug-resistant strains of malaria. Due to the either low abundance or low turn-over rate of the enzyme, obtaining artemisinin from both natural and synthetic sources is costly and laborious. In this in silico study, we strived to elucidate the substrate binding site specificities of the ADS, with the rational that unraveling enzyme features paves the way for enzyme engineering to increase synthesis rate. A homology model of the ADS from Artemisia annua L. was constructed based on the available crystal structure of the 5-epiaristolochene synthase (TEAS) and further analyzed with molecular dynamic simulations to determine residues forming the substrate recognition pocket. We also investigated the structural aspects of Mg 2+ binding. Results revealed DDYTD and NDLMT as metal-binding motifs in the putative active site gorge, which is composed of the D and H helixes and one loop region (aa519-532). Moreover, several representative residues including Tyr519, Asp444, Trp271, Asn443, Thr399, Arg262, Val292, Gly400 and Leu405, determine the FPP binding mode and its fate in terms of stereochemistry as well as the enzyme fidelity for the specific end product. These findings lead to inferences concerning key components of the ADS catalytic cavity, and provide evidence for the spatial localization of the FPP and Mg 2+ . Such detailed understanding will probably help to design an improved enzyme.

  4. Metal active site elasticity linked to activation of homocysteine in methionine synthases

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Koutmos, Markos; Pejchal, Robert; Bomer, Theresa M.

    2008-04-02

    Enzymes possessing catalytic zinc centers perform a variety of fundamental processes in nature, including methyl transfer to thiols. Cobalamin-independent (MetE) and cobalamin-dependent (MetH) methionine synthases are two such enzyme families. Although they perform the same net reaction, transfer of a methyl group from methyltetrahydrofolate to homocysteine (Hcy) to form methionine, they display markedly different catalytic strategies, modular organization, and active site zinc centers. Here we report crystal structures of zinc-replete MetE and MetH, both in the presence and absence of Hcy. Structural investigation of the catalytic zinc sites of these two methyltransferases reveals an unexpected inversion of zinc geometry uponmore » binding of Hcy and displacement of an endogenous ligand in both enzymes. In both cases a significant movement of the zinc relative to the protein scaffold accompanies inversion. These structures provide new information on the activation of thiols by zinc-containing enzymes and have led us to propose a paradigm for the mechanism of action of the catalytic zinc sites in these and related methyltransferases. Specifically, zinc is mobile in the active sites of MetE and MetH, and its dynamic nature helps facilitate the active site conformational changes necessary for thiol activation and methyl transfer.« less

  5. Gunnison sage-grouse lek site suitability modeling

    USGS Publications Warehouse

    Ouren, Douglas S.; Ignizio, Drew A.; Siders, Melissa; Childers, Theresa; Tucker, Karen; Seward, Nathan

    2014-01-01

    In order to better understand and protect species with minimal or decreasing populations, it is imperative to determine their actual existing population size. The focal species for this project is the Gunnison sage-grouse (GUSG), which became a proposed endangered species under the Endangered Species Act, thus confirming the need for better population estimates. Lek site counting during mating season has historically been the primary method for estimating population size since the grouse are very difficult to count at other times of the year. The objective of this project was to use historical data and available technology to identify additional potential lekking sites. This was done by determining areas throughout the study area that have the same landscape characteristics as those where known lekking activities occur. More accurate population counts could be the outcome of locating more lek sites. One of the remaining seven GUSG populations, the Crawford population (estimated at 128 individuals) exists in an area that includes the Gunnison Gorge National Conservation Area and the northern portion of the Black Canyon of the Gunnison National Park (our study area). While the Crawford population is small, it is still considered a self-sustaining population; the persistence and growth of this population directly contribute to genetic diversity conservation of this declining species. To date, only observational and anecdotal information about the Crawford population’s range, movements, and seasonal habitat use exist. From 1978 to the present, GUSG population monitoring has been accomplished through annual lek counts conducted each spring during GUSG mating season. Although this method has provided information on GUSG population trends, it is somewhat limited because counts are based only on known lekking sites and historically minimal efforts have been made to identify additional lek sites. To meet the objective of locating more potential lekking sites, we used a

  6. Nicotinamide Cofactors Suppress Active-Site Labeling of Aldehyde Dehydrogenases.

    PubMed

    Stiti, Naim; Chandrasekar, Balakumaran; Strubl, Laura; Mohammed, Shabaz; Bartels, Dorothea; van der Hoorn, Renier A L

    2016-06-17

    Active site labeling by (re)activity-based probes is a powerful chemical proteomic tool to globally map active sites in native proteomes without using substrates. Active site labeling is usually taken as a readout for the active state of the enzyme because labeling reflects the availability and reactivity of active sites, which are hallmarks for enzyme activities. Here, we show that this relationship holds tightly, but we also reveal an important exception to this rule. Labeling of Arabidopsis ALDH3H1 with a chloroacetamide probe occurs at the catalytic Cys, and labeling is suppressed upon nitrosylation and oxidation, and upon treatment with other Cys modifiers. These experiments display a consistent and strong correlation between active site labeling and enzymatic activity. Surprisingly, however, labeling is suppressed by the cofactor NAD(+), and this property is shared with other members of the ALDH superfamily and also detected for unrelated GAPDH enzymes with an unrelated hydantoin-based probe in crude extracts of plant cell cultures. Suppression requires cofactor binding to its binding pocket. Labeling is also suppressed by ALDH modulators that bind at the substrate entrance tunnel, confirming that labeling occurs through the substrate-binding cavity. Our data indicate that cofactor binding adjusts the catalytic Cys into a conformation that reduces the reactivity toward chloroacetamide probes.

  7. Quantum mechanical design of enzyme active sites.

    PubMed

    Zhang, Xiyun; DeChancie, Jason; Gunaydin, Hakan; Chowdry, Arnab B; Clemente, Fernando R; Smith, Adam J T; Handel, T M; Houk, K N

    2008-02-01

    The design of active sites has been carried out using quantum mechanical calculations to predict the rate-determining transition state of a desired reaction in presence of the optimal arrangement of catalytic functional groups (theozyme). Eleven versatile reaction targets were chosen, including hydrolysis, dehydration, isomerization, aldol, and Diels-Alder reactions. For each of the targets, the predicted mechanism and the rate-determining transition state (TS) of the uncatalyzed reaction in water is presented. For the rate-determining TS, a catalytic site was designed using naturalistic catalytic units followed by an estimation of the rate acceleration provided by a reoptimization of the catalytic site. Finally, the geometries of the sites were compared to the X-ray structures of related natural enzymes. Recent advances in computational algorithms and power, coupled with successes in computational protein design, have provided a powerful context for undertaking such an endeavor. We propose that theozymes are excellent candidates to serve as the active site models for design processes.

  8. Assessing soil heavy metal pollution in the water-level-fluctuation zone of the Three Gorges Reservoir, China.

    PubMed

    Ye, Chen; Li, Siyue; Zhang, Yulong; Zhang, Quanfa

    2011-07-15

    The water-level-fluctuation zone (WLFZ) between the elevations of 145-175 m in China's Three Gorges Reservoir has experienced a novel hydrological regime with half a year (May-September) exposed in summer and another half (October-April) submerged in winter. In September 2008 (before submergence) and June 2009 (after submergence), soil samples were collected in 12 sites in the WLFZ and heavy metals (Hg, As, Cr, Cd, Pb, Cu, Zn, Fe, and Mn) were determined. Enrichment factor (EF), factor analysis (FA), and factor analysis-multiple linear regression (FA-MLR) were employed for heavy metal pollution assessment, source identification, and source apportionment, respectively. Results demonstrate spatial variability in heavy metals before and after submergence and elements of As, Cd, Pb, Cu, and Zn are higher in the upper and low reaches. FA and FA-MLR reveal that As and Cd are the primary pollutants before submergence, and over 45% of As originates from domestic sewage and 59% of Cd from industrial wastes. After submergence, the major contaminants are Hg, Cd, and Pb, and traffic exhaust contributes approximately 81% to Hg and industrial effluent accounts about 36% and 73% for Cd and Pb, respectively. Our results suggest that increased shipping and industrial wastes have deposited large amounts of heavy metals which have been accumulated in the WLFZ during submergence period. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Beyond the gorge: Palaeoflood reconstruction from slackwater deposits in a range of physiographic settings in subtropical Australia

    NASA Astrophysics Data System (ADS)

    Lam, Daryl; Croke, Jacky; Thompson, Chris; Sharma, Ashneel

    2017-09-01

    The application of palaeoflood hydrology in Australia has been limited since its initial introduction > 30 years ago. This study adopts a regional, field-based approach to sampling slackwater deposits in a subtropical setting in southeast Queensland beyond the traditional arid setting. We explore the potential and challenges of using sites outside the traditional physiographical setting of bedrock gorges. Over 30 flood units were identified across different physiographical settings using a range of criteria. Evidence of charcoal-rich layers and palaeosol development assisted in the identification and separation of distinct flood units. The OSL-dated flood units are relatively young with two-thirds of the samples being < 1000 years old. The elevation of all flood units have resulted in estimated minimum discharges greater than the 1% annual exceedance probability. Although these are in the same order of gauged flood magnitudes, > 80% of them classified as 'extreme event'. This study opens up the renewed possibility of applying palaeoflood hydrology to more populated parts of Australia where the need for improved estimation of flood frequency and magnitude is now urgent in light of several extreme flood events. Preliminary contributions to improve the understanding between high magnitude floods and regional climatic drivers are also discussed. Recognised regional extreme floods generally coincide with La Niña and negative IPO phases, while tropical cyclones appear to be a key weather system in generating such large floods.

  10. Distribution, sedimentation, and bioavailability of particulate phosphorus in the mainstream of the Three Gorges Reservoir.

    PubMed

    Tang, Xianqiang; Wu, Min; Li, Rui

    2018-09-01

    The transportation and sedimentation of particulate phosphorus (PP) in a huge reservoir such as the Three Gorges Reservoir (TGR) are closely related to the phosphorus distribution characteristics and nutritional status of the water body. In this study, the PP distribution, sedimentation, and bioavailability in the mainstream section of the TGR were investigated through a field survey, indoor simulated settlement experiment, and historical data analysis. The results indicated that PP was the major component of the total phosphorus (TP) and that the Three Gorges Dam (TGD) trapped nearly 76.25% of suspended sediment (SS) and 75.35% of PP in the TGR, even during the flood season. A decline in flow velocity promoted the deposition of PP; additionally, PP concentrations gradually dropped from 0.35 mg/L in Chongqing to 0.02 mg/L in Zigui. The static PP sedimentation process adequately fitted a pseudo-second-order kinetic equation with a maximum correlation coefficient of 0.97. Moreover, more than half of the PP sedimentation process was achieved in less than 60 min for samples collected from the upper river reaches within simulated sedimentation process. The median particle size of SS and absolute value of the water column's zeta potential were negatively and positively related to the t 12 values of PP sedimentation, respectively. Compared with the concentration and particle size of SS obtained in the pre-TGR period, the values in the mainstream section of the TGR were lower. However, the TP and Fe/Al-P contents in SS increased several times. Due to the combined effects of flow velocity reduction and SS trapping, the water transparency and bioavailability of water column phosphorus were enhanced. Thus, the risk of water bloom outburst significantly increased when the impounded water level of 175 m in the TGR became the normal state. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Linkage between Three Gorges Dam impacts and the dramatic recessions in China’s largest freshwater lake, Poyang Lake

    PubMed Central

    Mei, Xuefei; Dai, Zhijun; Du, Jinzhou; Chen, Jiyu

    2015-01-01

    Despite comprising a small portion of the earth’s surface, lakes are vitally important for global ecosystem cycling. However, lake systems worldwide are extremely fragile, and many are shrinking due to changing climate and anthropogenic activities. Here, we show that Poyang Lake, the largest freshwater lake in China, has experienced a dramatic and prolonged recession, which began in late September of 2003. We further demonstrate that abnormally low levels appear during October, 28 days ahead of the normal initiation of the dry season, which greatly imperiled the lake’s wetland areas and function as an ecosystem for wintering waterbirds. An increase in the river-lake water level gradient induced by the Three Gorges Dam (TGD) altered the lake balance by inducing greater discharge into the Changjiang River, which is probably responsible for the current lake shrinkage. Occasional episodes of arid climate, as well as local sand mining, will aggravate the lake recession crisis. Although impacts of TGD on the Poyang Lake recession can be overruled by episodic extreme droughts, we argue that the average contributions of precipitation variation, human activities in the Poyang Lake catchment and TGD regulation to the Poyang Lake recession can be quantified as 39.1%, 4.6% and 56.3%, respectively. PMID:26657816

  12. Linkage between Three Gorges Dam impacts and the dramatic recessions in China’s largest freshwater lake, Poyang Lake

    NASA Astrophysics Data System (ADS)

    Mei, Xuefei; Dai, Zhijun; Du, Jinzhou; Chen, Jiyu

    2015-12-01

    Despite comprising a small portion of the earth’s surface, lakes are vitally important for global ecosystem cycling. However, lake systems worldwide are extremely fragile, and many are shrinking due to changing climate and anthropogenic activities. Here, we show that Poyang Lake, the largest freshwater lake in China, has experienced a dramatic and prolonged recession, which began in late September of 2003. We further demonstrate that abnormally low levels appear during October, 28 days ahead of the normal initiation of the dry season, which greatly imperiled the lake’s wetland areas and function as an ecosystem for wintering waterbirds. An increase in the river-lake water level gradient induced by the Three Gorges Dam (TGD) altered the lake balance by inducing greater discharge into the Changjiang River, which is probably responsible for the current lake shrinkage. Occasional episodes of arid climate, as well as local sand mining, will aggravate the lake recession crisis. Although impacts of TGD on the Poyang Lake recession can be overruled by episodic extreme droughts, we argue that the average contributions of precipitation variation, human activities in the Poyang Lake catchment and TGD regulation to the Poyang Lake recession can be quantified as 39.1%, 4.6% and 56.3%, respectively.

  13. Racemization of amino acids in fossil bones and teeth from the Olduvai Gorge region, Tanzania, East Africa

    NASA Astrophysics Data System (ADS)

    Bada, Jeffrey L.

    1981-10-01

    Investigation of amino acid racemization in fossil bones and teeth from the Olduvai Gorge region, Tanzania, indicates that aspartic acid racemization can be used to date samples which are less than ˜80,000-100,000 years old in this area. In older samples, significant secondary aspartic acid is apparently present and thus these samples have lower than expectedD/L aspartic acid ratios. Isoleucine in older samples, however, is apparently syngenetic with the samples, so the epimerization of isoleucine to alloisoleucine can be used with certain limitations to date fossil bones and teeth from the older stratigraphic sections in the Olduvai region.

  14. Active and regulatory sites of cytosolic 5'-nucleotidase.

    PubMed

    Pesi, Rossana; Allegrini, Simone; Careddu, Maria Giovanna; Filoni, Daniela Nicole; Camici, Marcella; Tozzi, Maria Grazia

    2010-12-01

    Cytosolic 5'-nucleotidase (cN-II), which acts preferentially on 6-hydroxypurine nucleotides, is essential for the survival of several cell types. cN-II catalyses both the hydrolysis of nucleotides and transfer of their phosphate moiety to a nucleoside acceptor through formation of a covalent phospho-intermediate. Both activities are regulated by a number of phosphorylated compounds, such as diadenosine tetraphosphate (Ap₄A), ADP, ATP, 2,3-bisphosphoglycerate (BPG) and phosphate. On the basis of a partial crystal structure of cN-II, we mutated two residues located in the active site, Y55 and T56. We ascertained that the ability to catalyse the transfer of phosphate depends on the presence of a bulky residue in the active site very close to the aspartate residue that forms the covalent phospho-intermediate. The molecular model indicates two possible sites at which adenylic compounds may interact. We mutated three residues that mediate interaction in the first activation site (R144, N154, I152) and three in the second (F127, M436 and H428), and found that Ap₄A and ADP interact with the same site, but the sites for ATP and BPG remain uncertain. The structural model indicates that cN-II is a homotetrameric protein that results from interaction through a specific interface B of two identical dimers that have arisen from interaction of two identical subunits through interface A. Point mutations in the two interfaces and gel-filtration experiments indicated that the dimer is the smallest active oligomerization state. Finally, gel-filtration and light-scattering experiments demonstrated that the native enzyme exists as a tetramer, and no further oligomerization is required for enzyme activation. © 2010 The Authors Journal compilation © 2010 FEBS.

  15. Water in the Active Site of Ketosteroid Isomerase

    PubMed Central

    Hanoian, Philip; Hammes-Schiffer, Sharon

    2011-01-01

    Classical molecular dynamics simulations were utilized to investigate the structural and dynamical properties of water in the active site of ketosteroid isomerase (KSI) to provide insight into the role of these water molecules in the enzyme-catalyzed reaction. This reaction is thought to proceed via a dienolate intermediate that is stabilized by hydrogen bonding with residues Tyr16 and Asp103. A comparative study was performed for the wild-type (WT) KSI and the Y16F, Y16S, and Y16F/Y32F/Y57F (FFF) mutants. These systems were studied with three different bound ligands: equilenin, which is an intermediate analog, and the intermediate states of two steroid substrates. Several distinct water occupation sites were identified in the active site of KSI for the WT and mutant systems. Three additional sites were identified in the Y16S mutant that were not occupied in WT KSI or the other mutants studied. The number of water molecules directly hydrogen bonded to the ligand oxygen was approximately two waters in the Y16S mutant, one water in the Y16F and FFF mutants, and intermittent hydrogen bonding of one water molecule in WT KSI. The molecular dynamics trajectories of the Y16F and FFF mutants reproduced the small conformational changes of residue 16 observed in the crystal structures of these two mutants. Quantum mechanical/molecular mechanical calculations of 1H NMR chemical shifts of the protons in the active site hydrogen-bonding network suggest that the presence of water in the active site does not prevent the formation of short hydrogen bonds with far-downfield chemical shifts. The molecular dynamics simulations indicate that the active site water molecules exchange much more frequently for WT KSI and the FFF mutant than for the Y16F and Y16S mutants. This difference is most likely due to the hydrogen-bonding interaction between Tyr57 and an active site water molecule that is persistent in the Y16F and Y16S mutants but absent in the FFF mutant and significantly less

  16. Water in the active site of ketosteroid isomerase.

    PubMed

    Hanoian, Philip; Hammes-Schiffer, Sharon

    2011-08-09

    Classical molecular dynamics simulations were utilized to investigate the structural and dynamical properties of water in the active site of ketosteroid isomerase (KSI) to provide insight into the role of these water molecules in the enzyme-catalyzed reaction. This reaction is thought to proceed via a dienolate intermediate that is stabilized by hydrogen bonding with residues Tyr16 and Asp103. A comparative study was performed for the wild-type (WT) KSI and the Y16F, Y16S, and Y16F/Y32F/Y57F (FFF) mutants. These systems were studied with three different bound ligands: equilenin, which is an intermediate analog, and the intermediate states of two steroid substrates. Several distinct water occupation sites were identified in the active site of KSI for the WT and mutant systems. Three additional sites were identified in the Y16S mutant that were not occupied in WT KSI or the other mutants studied. The number of water molecules directly hydrogen bonded to the ligand oxygen was approximately two in the Y16S mutant and one in the Y16F and FFF mutants, with intermittent hydrogen bonding of one water molecule in WT KSI. The molecular dynamics trajectories of the Y16F and FFF mutants reproduced the small conformational changes of residue 16 observed in the crystal structures of these two mutants. Quantum mechanical/molecular mechanical calculations of (1)H NMR chemical shifts of the protons in the active site hydrogen-bonding network suggest that the presence of water in the active site does not prevent the formation of short hydrogen bonds with far-downfield chemical shifts. The molecular dynamics simulations indicate that the active site water molecules exchange much more frequently for WT KSI and the FFF mutant than for the Y16F and Y16S mutants. This difference is most likely due to the hydrogen-bonding interaction between Tyr57 and an active site water molecule that is persistent in the Y16F and Y16S mutants but absent in the FFF mutant and significantly less probable

  17. Active Site Characterization of Proteases Sequences from Different Species of Aspergillus.

    PubMed

    Morya, V K; Yadav, Virendra K; Yadav, Sangeeta; Yadav, Dinesh

    2016-09-01

    A total of 129 proteases sequences comprising 43 serine proteases, 36 aspartic proteases, 24 cysteine protease, 21 metalloproteases, and 05 neutral proteases from different Aspergillus species were analyzed for the catalytically active site residues using MEROPS database and various bioinformatics tools. Different proteases have predominance of variable active site residues. In case of 24 cysteine proteases of Aspergilli, the predominant active site residues observed were Gln193, Cys199, His364, Asn384 while for 43 serine proteases, the active site residues namely Asp164, His193, Asn284, Ser349 and Asp325, His357, Asn454, Ser519 were frequently observed. The analysis of 21 metalloproteases of Aspergilli revealed Glu298 and Glu388, Tyr476 as predominant active site residues. In general, Aspergilli species-specific active site residues were observed for different types of protease sequences analyzed. The phylogenetic analysis of these 129 proteases sequences revealed 14 different clans representing different types of proteases with diverse active site residues.

  18. Half-of-the-sites reactivity of outer-membrane phospholipase A against an active-site-directed inhibitor.

    PubMed

    Ubarretxena-Belandia, I; Cox, R C; Dijkman, R; Egmond, M R; Verheij, H M; Dekker, N

    1999-03-01

    The reaction of a novel active-site-directed phospholipase A1 inhibitor with the outer-membrane phospholipase A (OMPLA) was investigated. The inhibitor 1-p-nitrophenyl-octylphosphonate-2-tridecylcarbamoyl-3-et hanesulfonyl -amino-3-deoxy-sn-glycerol irreversibly inactivated OMPLA. The inhibition reaction did not require the cofactor calcium or an unprotonated active-site His142. The inhibition of the enzyme solubilized in hexadecylphosphocholine micelles was characterized by a rapid (t1/2 = 20 min) and complete loss of enzymatic activity, concurrent with the covalent modification of 50% of the active-site serines, as judged from the amount of p-nitrophenolate (PNP) released. Modification of the remaining 50% occurred at a much lower rate, indicative of half-of-the-sites reactivity against the inhibitor of this dimeric enzyme. Inhibition of monomeric OMPLA solubilized in hexadecyl-N,N-dimethyl-1-ammonio-3-propanesulfonate resulted in an equimolar monophasic release of PNP, concurrent with the loss of enzymatic activity (t1/2 = 14 min). The half-of-the-sites reactivity is discussed in view of the dimeric nature of this enzyme.

  19. Physical activity and cancer risk: dose-response and cancer, all sites and site-specific.

    PubMed

    Thune, I; Furberg, A S

    2001-06-01

    The association between physical activity and overall and site-specific cancer risk is elaborated in relation to whether any observed dose-response association between physical activity and cancer can be interpreted in terms of how much physical activity (type, intensity, duration, frequency) is needed to influence site- and gender-specific cancer risk. Observational studies were reviewed that have examined the independent effect of the volume of occupational physical activity (OPA) and/or leisure time physical activity (LPA) on overall and site-specific cancer risk. The evidence of cohort and case-control studies suggests that both leisure time and occupational physical activity protect against overall cancer risk, with a graded dose-response association suggested in both sexes. Confounding effects such as diet, body weight, and parity are often included as a covariate in the analyses, with little influence on the observed associations. A crude graded inverse dose-response association was observed between physical activity and colon cancer in 48 studies including 40,674 colon/colorectal cancer cases for both sexes. A dose-response effect of physical activity on colon cancer risk was especially observed, when participation in activities of at least moderate activity (>4.5 MET) and demonstrated by activities expressed as MET-hours per week. An observed inverse association with a dose-response relationship between physical activity and breast cancer was also identified in the majority of the 41 studies including 108,031 breast cancer cases. The dose-response relationship was in particular observed in case-control studies and supported by observations in cohort studies when participation in activities of at least moderate activity (>4.5 MET) and demonstrated by activities expressed by MET-hours per week. This association between physical activity and breast cancer risk is possibly dependent on age at exposure, age at diagnosis, menopausal status and other effect

  20. Active Site Metal Occupancy and Cyclic Di-GMP Phosphodiesterase Activity of Thermotoga maritima HD-GYP.

    PubMed

    Miner, Kyle D; Kurtz, Donald M

    2016-02-16

    HD-GYPs make up a subclass of the metal-dependent HD phosphohydrolase superfamily and catalyze conversion of cyclic di(3',5')-guanosine monophosphate (c-di-GMP) to 5'-phosphoguanylyl-(3'→5')-guanosine (pGpG) and GMP. Until now, the only reported crystal structure of an HD-GYP that also exhibits c-di-GMP phosphodiesterase activity contains a His/carboxylate ligated triiron active site. However, other structural and phylogenetic correlations indicate that some HD-GYPs contain dimetal active sites. Here we provide evidence that an HD-GYP c-di-GMP phosphodiesterase, TM0186, from Thermotoga maritima can accommodate both di- and trimetal active sites. We show that an as-isolated iron-containing TM0186 has an oxo/carboxylato-bridged diferric site, and that the reduced (diferrous) form is necessary and sufficient to catalyze conversion of c-di-GMP to pGpG, but that conversion of pGpG to GMP requires more than two metals per active site. Similar c-di-GMP phosphodiesterase activities were obtained with divalent iron or manganese. On the basis of activity correlations with several putative metal ligand residue variants and molecular dynamics simulations, we propose that TM0186 can accommodate both di- and trimetal active sites. Our results also suggest that a Glu residue conserved in a subset of HD-GYPs is required for formation of the trimetal site and can also serve as a labile ligand to the dimetal site. Given the anaerobic growth requirement of T. maritima, we suggest that this HD-GYP can function in vivo with either divalent iron or manganese occupying di- and trimetal sites.

  1. Vegetation during UMBI and deposition of Tuff IF at Olduvai Gorge, Tanzania (ca. 1.8 Ma) based on phytoliths and plant remains.

    PubMed

    Albert, Rosa Maria; Bamford, Marion K

    2012-08-01

    As part of ongoing research at Olduvai Gorge, Tanzania, to determine the detailed paleoenvironmental setting during Bed I and Bed II times and occupation of the basin by early hominins, we present the results of phytolith analyses of Tuff IF which is the uppermost unit of Bed I. Phytoliths were identified in most of the levels and localities on the eastern paleolake margin, but there are not always sufficient numbers of identifiable morphologies to infer the specific type of vegetation due to dissolution. Some surge surfaces and reworked tuff surfaces were vegetated between successive ash falls, as indicated by root-markings and the presence of a variety of phytolith morphotypes. Dicotyledonous wood/bark types were dominant except at the FLK N site just above Tuff IF when monocots are dominant and for the palm-dominated sample from the reworked channel cutting down into Tuff IF at FLK N. The area between the two fault scarps bounding the HWK Compartment, approximately 1 km wide, was vegetated at various time intervals between some of the surges and during the reworking of the Tuff. By lowermost Bed II times the eastern margin was fully vegetated again. Climate and tectonic activity probably controlled the fluctuating lake levels but locally the paleorelief and drainage were probably the controlling factors for the vegetation changes. These data support a scenario of small groups of hominins making brief visits to the paleolake during uppermost Bed I times, followed by a more desirable vegetative environment during lowermost Bed II times. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Acoustic Emission Parameters of Three Gorges Sandstone during Shear Failure

    NASA Astrophysics Data System (ADS)

    Xu, Jiang; Liu, Yixin; Peng, Shoujian

    2016-12-01

    In this paper, an experimental investigation of sandstone samples from the Three Gorges during shear failure was conducted using acoustic emission (AE) and direct shear tests. The AE count rate, cumulative AE count, AE energy, and amplitude of the sandstone samples were determined. Then, the relationships among the AE signals and shearing behaviors of the samples were analyzed in order to detect micro-crack initiation and propagation and reflect shear failure. The results indicated that both the shear strength and displacement exhibited a logarithmic relationship with the displacement rate at peak levels of stress. In addition, the various characteristics of the AE signals were apparent in various situations. The AE signals corresponded with the shear stress under different displacement rates. As the displacement rate increased, the amount of accumulative damage to each specimen decreased, while the AE energy peaked earlier and more significantly. The cumulative AE count primarily increased during the post-peak period. Furthermore, the AE count rate and amplitude exhibited two peaks during the peak shear stress period due to crack coalescence and rock bridge breakage. These isolated cracks later formed larger fractures and eventually caused ruptures.

  3. Diffusional correlations among multiple active sites in a single enzyme.

    PubMed

    Echeverria, Carlos; Kapral, Raymond

    2014-04-07

    Simulations of the enzymatic dynamics of a model enzyme containing multiple substrate binding sites indicate the existence of diffusional correlations in the chemical reactivity of the active sites. A coarse-grain, particle-based, mesoscopic description of the system, comprising the enzyme, the substrate, the product and solvent, is constructed to study these effects. The reactive and non-reactive dynamics is followed using a hybrid scheme that combines molecular dynamics for the enzyme, substrate and product molecules with multiparticle collision dynamics for the solvent. It is found that the reactivity of an individual active site in the multiple-active-site enzyme is reduced substantially, and this effect is analyzed and attributed to diffusive competition for the substrate among the different active sites in the enzyme.

  4. [Impact of periodical flooding-drying on nitrification and ammonia oxidizers in hydro-fluctuation belt of the Three Gorges Reservoir].

    PubMed

    Guo, Jia; Jiang, Xianjun; Zhou, Xue; Meng, Yao; Jia, Zhongjun

    2016-06-04

    This study was aimed to elucidate the effect of periodic flooding-drying to ecological processes of ammonia oxidizers in the hydro-fluctuation belt of the Three Gorges Reservoir. Soil samples were collected at thee altitudes in regions of Wanzhou, Fengdu and Changshou, representing 8, 5 and 0 times floodingdrying management, respectively. Soil physiochemical properties were analyzed and microcosms were constructed to monitor nitrification activity by fertilizing soils with ammonium substrate. Real-time PCR was used to quantify the population size of ammonia-oxidizing archaea (AOA) and bacteria (AOB). DGGE fingerprints and clone libraries were conducted to study the shift of AOA and AOB compositions in nitrifying soils. Among the physiochemical characteristics of the soils, soil organic matter and total phosphates increased along with cycle increasing. After incubation for 13 days, the net nitrification rates of the samples with 8 cycles exceeded those with 5 cycles. The quantities of both AOA and AOB have increased during the incubation. Phylogenetic analysis showed that AOA were placed within the soil group 1.1b and soil group 1.1a, while bacterial ammonia oxidizers were closely related to Nitrosospira and Cluster 0. Periodical flooding-drying increased soil organic matter, enhanced soil nitrification activity and likely played important roles in shaping community structures of soil ammonia oxidizers.

  5. Hydraulic characteristics of the New River in the New River Gorge National River, West Virginia

    USGS Publications Warehouse

    Wiley, J.B.; Appel, David H.

    1989-01-01

    Traveltime, dispersion, water-surface and streambed profiles, and cross-section data were collected for use in application of flow and solute-transport models to the New River in the New River Gorge National River, West Virginia. Dye clouds subjected to increasing and decreasing flow rates (unsteady flow) showed that increasing flows shorten the cloud and decreasing flows lengthen the cloud. After the flow rate was changed and the flow was again steady, traveltime and dispersion characteristics were determined by the new rate of flow. Seven stage/streamflow relations identified the general changes of stream geometry throughout the study reach. Channel cross sections were estimated for model input. Low water and streambed profiles were developed from surveyed water surface elevations and water depths. (USGS)

  6. Direct instrumental identification of catalytically active surface sites

    NASA Astrophysics Data System (ADS)

    Pfisterer, Jonas H. K.; Liang, Yunchang; Schneider, Oliver; Bandarenka, Aliaksandr S.

    2017-09-01

    The activity of heterogeneous catalysts—which are involved in some 80 per cent of processes in the chemical and energy industries—is determined by the electronic structure of specific surface sites that offer optimal binding of reaction intermediates. Directly identifying and monitoring these sites during a reaction should therefore provide insight that might aid the targeted development of heterogeneous catalysts and electrocatalysts (those that participate in electrochemical reactions) for practical applications. The invention of the scanning tunnelling microscope (STM) and the electrochemical STM promised to deliver such imaging capabilities, and both have indeed contributed greatly to our atomistic understanding of heterogeneous catalysis. But although the STM has been used to probe and initiate surface reactions, and has even enabled local measurements of reactivity in some systems, it is not generally thought to be suited to the direct identification of catalytically active surface sites under reaction conditions. Here we demonstrate, however, that common STMs can readily map the catalytic activity of surfaces with high spatial resolution: we show that by monitoring relative changes in the tunnelling current noise, active sites can be distinguished in an almost quantitative fashion according to their ability to catalyse the hydrogen-evolution reaction or the oxygen-reduction reaction. These data allow us to evaluate directly the importance and relative contribution to overall catalyst activity of different defects and sites at the boundaries between two materials. With its ability to deliver such information and its ready applicability to different systems, we anticipate that our method will aid the rational design of heterogeneous catalysts.

  7. Miocene Basaltic Lava Flows and Dikes of the Intervening Area Between Picture Gorge and Steens Basalt of the CRBG, Eastern Oregon

    NASA Astrophysics Data System (ADS)

    Cahoon, E. B.; Streck, M. J.

    2016-12-01

    Mid-Miocene basaltic lavas and dikes are exposed in the area between the southern extent of the Picture Gorge Basalt (PGB) and the northern extent of Steens Basalt in a wide corridor of the Malheur National Forest, eastern Oregon. An approximate mid-Miocene age of sampled basaltic units is indicated by stratigraphic relationships to the 16 Ma Dinner Creek Tuff. Lavas provide an opportunity to extend and/or revise distribution areas of either CRBG unit and explore the petrologic transition between them. The PGB and the Steens Basalt largely represent geochemically distinct tholeiitic units of the CRBG; although each unit displays internal complexity. Lavas of PGB are relatively primitive (MgO 5-9 wt.%) while Steens Basalt ranges in MgO from >9 to 3 wt.% but both units are commonly coarsely porphyritic. Conversely, Steens Basalt compositions are on average more enriched in highly incompatible elements (e.g. Rb, Th) and relatively enriched in the lesser incompatible elements (e.g. Y, Yb) compared to the Picture Gorge basalts. These compositional signatures produce inclined and flat patterns on mantle-normalized incompatible trace element plots but with similar troughs and spikes, respectively. New compositional data from our study area indicate basaltic lavas can be assigned as PGB lava flows and dikes, and also to a compositional group chemically distinct between Steens Basalt and PGB. Distribution of lava flows with PGB composition extend this CRBG unit significantly south/southeast closing the exposure gap between PGB and Steens Basalt. We await data that match Steens Basalt compositions but basaltic lavas with petrographic features akin to Steens Basalt have been identified in the study area. Lavas of the transitional unit share characteristics with Upper Steens and Picture Gorge basalt types, but identify a new seemingly unique composition. This composition is slightly more depleted in the lesser incompatible elements (i.e. steeper pattern) on mantle normalized

  8. The identity of the active site of oxalate decarboxylase and the importance of the stability of active-site lid conformations1

    PubMed Central

    Just, Victoria J.; Burrell, Matthew R.; Bowater, Laura; McRobbie, Iain; Stevenson, Clare E. M.; Lawson, David M.; Bornemann, Stephen

    2007-01-01

    Oxalate decarboxylase (EC 4.1.1.2) catalyses the conversion of oxalate into carbon dioxide and formate. It requires manganese and, uniquely, dioxygen for catalysis. It forms a homohexamer and each subunit contains two similar, but distinct, manganese sites termed sites 1 and 2. There is kinetic evidence that only site 1 is catalytically active and that site 2 is purely structural. However, the kinetics of enzymes with mutations in site 2 are often ambiguous and all mutant kinetics have been interpreted without structural information. Nine new site-directed mutants have been generated and four mutant crystal structures have now been solved. Most mutants targeted (i) the flexibility (T165P), (ii) favoured conformation (S161A, S164A, D297A or H299A) or (iii) presence (Δ162–163 or Δ162–164) of a lid associated with site 1. The kinetics of these mutants were consistent with only site 1 being catalytically active. This was particularly striking with D297A and H299A because they disrupted hydrogen bonds between the lid and a neighbouring subunit only when in the open conformation and were distant from site 2. These observations also provided the first evidence that the flexibility and stability of lid conformations are important in catalysis. The deletion of the lid to mimic the plant oxalate oxidase led to a loss of decarboxylase activity, but only a slight elevation in the oxalate oxidase side reaction, implying other changes are required to afford a reaction specificity switch. The four mutant crystal structures (R92A, E162A, Δ162–163 and S161A) strongly support the hypothesis that site 2 is purely structural. PMID:17680775

  9. Role of active site rigidity in activity: MD simulation and fluorescence study on a lipase mutant.

    PubMed

    Kamal, Md Zahid; Mohammad, Tabrez Anwar Shamim; Krishnamoorthy, G; Rao, Nalam Madhusudhana

    2012-01-01

    Relationship between stability and activity of enzymes is maintained by underlying conformational flexibility. In thermophilic enzymes, a decrease in flexibility causes low enzyme activity while in less stable proteins such as mesophiles and psychrophiles, an increase in flexibility is associated with enhanced enzyme activity. Recently, we identified a mutant of a lipase whose stability and activity were enhanced simultaneously. In this work, we probed the conformational dynamics of the mutant and the wild type lipase, particularly flexibility of their active site using molecular dynamic simulations and time-resolved fluorescence techniques. In contrast to the earlier observations, our data show that active site of the mutant is more rigid than wild type enzyme. Further investigation suggests that this lipase needs minimal reorganization/flexibility of active site residues during its catalytic cycle. Molecular dynamic simulations suggest that catalytically competent active site geometry of the mutant is relatively more preserved than wild type lipase, which might have led to its higher enzyme activity. Our study implies that widely accepted positive correlation between conformation flexibility and enzyme activity need not be stringent and draws attention to the possibility that high enzyme activity can still be accomplished in a rigid active site and stable protein structures. This finding has a significant implication towards better understanding of involvement of dynamic motions in enzyme catalysis and enzyme engineering through mutations in active site.

  10. Portland, Mount Hood, & Columbia River Gorge, Oregon, Perspective View

    NASA Technical Reports Server (NTRS)

    2004-01-01

    Portland, the largest city in Oregon, is located on the Columbia River at the northern end of the Willamette Valley. On clear days, Mount Hood highlights the Cascade Mountains backdrop to the east. The Columbia is the largest river in the American Northwest and is navigable up to and well beyond Portland. It is also the only river to fully cross the Cascade Range, and has carved the Columbia River Gorge, which is seen in the left-central part of this view. A series of dams along the river, at topographically favorable sites, provide substantial hydroelectric power to the region.

    This perspective view was generated using topographic data from the Shuttle Radar Topography Mission (SRTM), a Landsat satellite image, and a false sky. Topographic expression is vertically exaggerated two times.

    Landsat has been providing visible and infrared views of the Earth since 1972. SRTM elevation data substantially help in analyzing Landsat images by revealing the third dimension of Earth's surface, topographic height. The Landsat archive is managed by the U.S. Geological Survey's Eros Data Center (USGS EDC).

    Elevation data used in this image were acquired by the Shuttle Radar Topography Mission (SRTM) aboard the Space Shuttle Endeavour, launched on February 11, 2000. SRTM used the same radar instrument that comprised the Spaceborne Imaging Radar-C/X-Band Synthetic Aperture Radar (SIR-C/X-SAR) that flew twice on the Space Shuttle Endeavour in 1994. SRTM was designed to collect three-dimensional measurements of the Earth's surface. To collect the 3-D data, engineers added a 60-meter-long (200-foot) mast, installed additional C-band and X-band antennas, and improved tracking and navigation devices. The mission is a cooperative project between the National Aeronautics and Space Administration (NASA), the National Geospatial-Intelligence Agency (NGA) of the U.S. Department of Defense (DoD), and the German and Italian space agencies. It is managed by NASA's Jet

  11. Estimating the impacts of reservoir elevation changes on kokanee emergence in flaming Gorge Reservoir, Wyoming-Utah

    USGS Publications Warehouse

    Modde, T.; Jeric, R.J.; Hubert, W.A.; Gipson, R.D.

    1997-01-01

    Flaming Gorge Reservoir, like many western North American reservoirs, is managed to release water during the winter months to allow for water storage associated with melting snow and rain during spring. Decreases in reservoir elevation during winter can cause mortalities of kokanee Oncorhynchus nerka spawned along the shoreline the previous fall. This study compared data on depth distribution of embryos and depth-adjusted survival to estimate the relative survival of emergent kokanee at different depths and the effect of winter drawdown on the proportion of deposited eggs that survive to emergence. Estimates of decreases in kokanee survival to emergence were 8.3% and 38.1% for reservoir elevation reductions of 1.0 m and 5.0 m, respectively.

  12. 40 CFR 61.154 - Standard for active waste disposal sites.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 8 2011-07-01 2011-07-01 false Standard for active waste disposal... for Asbestos § 61.154 Standard for active waste disposal sites. Each owner or operator of an active... visible emissions to the outside air from any active waste disposal site where asbestos-containing waste...

  13. 40 CFR 61.154 - Standard for active waste disposal sites.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 9 2013-07-01 2013-07-01 false Standard for active waste disposal... for Asbestos § 61.154 Standard for active waste disposal sites. Each owner or operator of an active... visible emissions to the outside air from any active waste disposal site where asbestos-containing waste...

  14. 40 CFR 61.154 - Standard for active waste disposal sites.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 9 2012-07-01 2012-07-01 false Standard for active waste disposal... for Asbestos § 61.154 Standard for active waste disposal sites. Each owner or operator of an active... visible emissions to the outside air from any active waste disposal site where asbestos-containing waste...

  15. 40 CFR 61.154 - Standard for active waste disposal sites.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 8 2010-07-01 2010-07-01 false Standard for active waste disposal... for Asbestos § 61.154 Standard for active waste disposal sites. Each owner or operator of an active... visible emissions to the outside air from any active waste disposal site where asbestos-containing waste...

  16. 40 CFR 61.154 - Standard for active waste disposal sites.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 9 2014-07-01 2014-07-01 false Standard for active waste disposal... for Asbestos § 61.154 Standard for active waste disposal sites. Each owner or operator of an active... visible emissions to the outside air from any active waste disposal site where asbestos-containing waste...

  17. The littoral zone in the Three Gorges Reservoir, China: challenges and opportunities.

    PubMed

    Yuan, Xing-zhong; Zhang, Yue-wei; Liu, Hong; Xiong, Sen; Li, Bo; Deng, Wei

    2013-10-01

    For flood control purpose, the water level of the Three Gorges Reservoir (TGR) varies significantly. The annual reservoir surface elevation amplitude is about 30 m behind the dam. Filling of the reservoir has created about 349 km(2) of newly flooded riparian zone. The average flooding period lasts for more than 6 months, from mid-October to late April. The dam and its associated reservoir provide flood control, power generation, and navigation, but there are also many environmental challenges. The littoral zone is the important part of the TGR, once its eco-health and stability are damaged,which will directly endanger the ecological safety of the whole reservoir area and even the Yangtze River Basin. So, understanding the great ecological opportunities which are hidden in littoral zone of TGR (LZTGR) and putting forward approaches to solve the environmental problems are very important. LZTGR involves a wide field of problems, such as the landslides, potential water pollution, soil erosion, biodiversity loss, land cover changes, and other issues. The Three Gorges dam (TGD) is a major trigger of environmental change in the Yangtze River. The landslides, water quality, soil erosion, loss of biodiversity, dam operation, and challenge for land use are closely interrelated across spatial and temporal scales. Therefore, the ecological and environmental impacts caused by TGD are necessarily complex and uncertain. LZTGR is not only a great environmental challenge but also an ecological opportunity for us. In fact, LZTGR is an important structural unit of TGR ecosystem and has special ecosystem services function. Vegetation growing in LZTGR is therefore a valuable resource due to accumulation of carbon and nutrients. Everyone thinks that the ecological approach to the problem is needed. If properly designed, dike-pond systems, littoral woods systems, and re-created waterfowl habitats will have the capacity to capture nutrients from uplands and obstruct soil erosion

  18. 10 CFR 63.16 - Review of site characterization activities. 2

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... which such activities are carried out and to observe excavations, borings, and in situ tests, as they... IN A GEOLOGIC REPOSITORY AT YUCCA MOUNTAIN, NEVADA Licenses Preapplication Review § 63.16 Review of site characterization activities. 2 2 In addition to the review of site characterization activities...

  19. 10 CFR 63.16 - Review of site characterization activities. 2

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... which such activities are carried out and to observe excavations, borings, and in situ tests, as they... IN A GEOLOGIC REPOSITORY AT YUCCA MOUNTAIN, NEVADA Licenses Preapplication Review § 63.16 Review of site characterization activities. 2 2 In addition to the review of site characterization activities...

  20. 10 CFR 63.16 - Review of site characterization activities. 2

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... which such activities are carried out and to observe excavations, borings, and in situ tests, as they... IN A GEOLOGIC REPOSITORY AT YUCCA MOUNTAIN, NEVADA Licenses Preapplication Review § 63.16 Review of site characterization activities. 2 2 In addition to the review of site characterization activities...

  1. 10 CFR 63.16 - Review of site characterization activities. 2

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... which such activities are carried out and to observe excavations, borings, and in situ tests, as they... IN A GEOLOGIC REPOSITORY AT YUCCA MOUNTAIN, NEVADA Licenses Preapplication Review § 63.16 Review of site characterization activities. 2 2 In addition to the review of site characterization activities...

  2. 10 CFR 63.16 - Review of site characterization activities. 2

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... which such activities are carried out and to observe excavations, borings, and in situ tests, as they... IN A GEOLOGIC REPOSITORY AT YUCCA MOUNTAIN, NEVADA Licenses Preapplication Review § 63.16 Review of site characterization activities. 2 2 In addition to the review of site characterization activities...

  3. An evaluation of the inhibition of human butyrylcholinesterase and acetylcholinesterase by the organophosphate chlorpyrifos oxon

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shenouda, Josephine; Green, Paula; Sultatos, Lester, E-mail: sultatle@umdnj.ed

    2009-12-01

    Acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8) are enzymes that belong to the superfamily of alpha/beta-hydrolase fold proteins. While they share many characteristics, they also possess many important differences. For example, whereas they have about 54% amino acid sequence identity, the active site gorge of acetylcholinesterase is considerably smaller than that of butyrylcholinesterase. Moreover, both have been shown to display simple and complex kinetic mechanisms, depending on the particular substrate examined, the substrate concentration, and incubation conditions. In the current study, incubation of butyrylthiocholine in a concentration range of 0.005-3.0 mM, with 317 pM human butyrylcholinesterase in vitro, resulted inmore » rates of production of thiocholine that were accurately described by simple Michaelis-Menten kinetics, with a K{sub m} of 0.10 mM. Similarly, the inhibition of butyrylcholinesterase in vitro by the organophosphate chlorpyrifos oxon was described by simple Michaelis-Menten kinetics, with a k{sub i} of 3048 nM{sup -1} h{sup -1}, and a K{sub D} of 2.02 nM. In contrast to inhibition of butyrylcholinesterase, inhibition of human acetylcholinesterase by chlorpyrifos oxon in vitro followed concentration-dependent inhibition kinetics, with the k{sub i} increasing as the inhibitor concentration decreased. Chlorpyrifos oxon concentrations of 10 and 0.3 nM gave k{sub i}s of 1.2 and 19.3 nM{sup -1} h{sup -1}, respectively. Although the mechanism of concentration-dependent inhibition kinetics is not known, the much smaller, more restrictive active site gorge of acetylcholinesterase almost certainly plays a role. Similarly, the much larger active site gorge of butyrylcholinesterase likely contributes to its much greater reactivity towards chlorpyrifos oxon, compared to acetylcholinesterase.« less

  4. Analysis of the reaction of carbachol with acetylcholinesterase using thioflavin T as a coupled fluorescence reporter.

    PubMed

    Rosenberry, Terrone L; Sonoda, Leilani K; Dekat, Sarah E; Cusack, Bernadette; Johnson, Joseph L

    2008-12-09

    Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acylated enzyme intermediate is produced. Carbamates are very poor substrates that, like other AChE substrates, form an initial enzyme-substrate complex with free AChE (E) and proceed to an acylated enzyme intermediate (EC), which is then hydrolyzed. However, the hydrolysis of EC is slow enough to resolve the acylation and deacylation steps on the catalytic pathway. Here, we focus on the reaction of carbachol (carbamoylcholine) with AChE. The kinetics and thermodynamics of this reaction are of special interest because carbachol is an isosteric analogue of the physiological substrate acetylcholine. We show that the reaction can be monitored with thioflavin T as a fluorescent reporter group. The fluorescence of thioflavin T is strongly enhanced when it binds to the P-site of AChE, and this fluorescence is partially quenched when a second ligand binds to the A-site to form a ternary complex. Analysis of the fluorescence reaction profiles was challenging because four thermodynamic parameters and two fluorescence coefficients were fitted from the combined data both for E and for EC. Respective equilibrium dissociation constants of 6 and 26 mM were obtained for carbachol binding to the A- and P-sites in E and of 2 and 32 mM for carbachol binding to the A- and P-sites in EC. These constants for the binding of carbachol to the P-site are about an order of magnitude larger (i.e., indicating lower affinity) than previous estimates for the binding of acetylthiocholine to the P-site.

  5. Analysis of the reaction of carbachol with acetylcholinesterase with thioflavin T as a coupled fluorescence reporter†

    PubMed Central

    Rosenberry, Terrone L.; Sonoda, Leilani K.; Dekat, Sarah E.; Cusack, Bernadette; Johnson, Joseph L.

    2009-01-01

    Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acylated enzyme intermediate is produced. Carbamates are very poor substrates that, like other AChE substrates, form an initial enzyme-substrate complex with free AChE (E) and proceed to an acylated enzyme intermediate (EC) which is then hydrolyzed. However, the hydrolysis of EC is slow enough to resolve the acylation and deacylation steps on the catalytic pathway. Here we focus on the reaction of carbachol (carbamoylcholine) with AChE. The kinetics and thermodynamics of this reaction are of special interest because carbachol is an isosteric analog of the physiological substrate acetylcholine. We show that the reaction can be monitored with thioflavin T as a fluorescent reporter group. The fluorescence of thioflavin T is strongly enhanced when it binds to the P-site of AChE, and this fluorescence is partially quenched when a second ligand binds to the A-site to form a ternary complex. Analysis of the fluorescence reaction profiles was challenging, because four thermodynamic parameters and two fluorescence coefficients were fitted from the combined data both for E and for EC. Respective equilibrium dissociation constants of 6 and 26 mM were obtained for carbachol binding to the A- and P-sites in E and of 2 and 32 mM for carbachol binding to the A- and P-sites in EC. These constants for the binding of carbachol to the P-site are about an order of magnitude larger (i.e., indicating lower affinity) than previous estimates for the binding of acetylthiocholine to the P-site. PMID:19006330

  6. Dissolved-oxygen depletion and other effects of storing water in Flaming Gorge Reservoir, Wyoming and Utah

    USGS Publications Warehouse

    Bolke, E.L.

    1979-01-01

    The circulation of water in Flaming Gorge Reservoir is caused chiefly by insolation, inflow-outflow relationships, and wind, which is significant due to the geographical location of the reservoir. During 1970-75, there was little annual variation in the thickness, dissolved oxygen, and specific conductance of the hypolimnion near Flaming Gorge Dam. Depletion of dissolved oxygen occurred simultaneously in the bottom waters of both tributary arms in the upstream part of the reservoir and was due to reservoir stratification. Anaerobic conditions in the bottom water during summer stratification eventually results in a metalimnetic oxygen minimum in the reservoir.The depletion of flow in the river below Flaming Gorge Dam due to evaporation and bank storage in the reservoir for the 1963-75 period was 1,320 cubic hectometers, and the increase of dissolved-solids load in the river was 1,947,000 metric tons. The largest annual variations in dissolved-solids concentration in the river was about 600 milligrams per liter before closure of the dam and about 200 milligrams per liter after closure. The discharge weighted-average dissolved-solids concentration for the 5 years prior to closure was 386 milligrams per liter and 512 milligrams per liter after closure. The most significant changes in the individual dissolved-ion loads in the river during 1973-75 were the increase in sulfate (0.46 million metric tons), which was probably derived from the solution of gypsum, and the decrease in bicarbonate (0.39 million metric tons), which can be attributed to chemical precipitation.The maximum range in temperature in the Green River below the reservoir prior to closure of the dam in 1962 was from 0°C in winter to 21°C in summer. After closure until 1970 the temperature ranged from 2° to 12°C, but since 1970 the range has been from 4° to 9°C.The maximum range in temperature in the Green River below the reservoir prior to closure of the dam in 1962 was from 0°C in winter to 21

  7. Advancing Analysis of Spatio-Temporal Variations of Soil Nutrients in the Water Level Fluctuation Zone of China’s Three Gorges Reservoir Using Self-Organizing Map

    PubMed Central

    Ye, Chen; Li, Siyue; Yang, Yuyi; Shu, Xiao; Zhang, Jiaquan; Zhang, Quanfa

    2015-01-01

    The ~350 km2 water level fluctuation zone (WLFZ) in the Three Gorges Reservoir (TGR) of China, situated at the intersection of terrestrial and aquatic ecosystems, experiences a great hydrological change with prolonged winter inundation. Soil samples were collected in 12 sites pre- (September 2008) and post submergence (June 2009) in the WLFZ and analyzed for soil nutrients. Self-organizing map (SOM) and statistical analysis including multi-way ANOVA, paired-T test, and stepwise least squares multiple regression were employed to determine the spatio-temporal variations of soil nutrients in relation to submergence, and their correlations with soil physical characteristics. Results showed significant spatial variability in nutrients along ~600 km long shoreline of the TGR before and after submergence. There were higher contents of organic matter, total nitrogen (TN), and nitrate (NO3-) in the lower reach and total phosphorus (TP) in the upper reach that were primarily due to the spatial variations in soil particle size composition and anthropogenic activities. Submergence enhanced soil available potassium (K), while significantly decreased soil N, possibly due to the alterations of soil particle size composition and increase in soil pH. In addition, SOM analysis determined important roles of soil pH value, bulk density, soil particle size (i.e., silt and sand) and nutrients (TP, TK, and AK) on the spatial and temporal variations in soil quality. Our results suggest that urban sewage and agricultural runoffs are primary pollutants that affect soil nutrients in the WLFZ of TGR. PMID:25789612

  8. [Spatial distribution of mercury in soils of a typical small agricultural watershed in the Three Gorges Reservoir region].

    PubMed

    Wang, Ya; Zhao, Zheng; Mu, Zhi-jian; Wang, Dlng-yong; Yu, Ya-wei

    2015-01-01

    To understand the mercury (Hg) pollution level and the corresponding ecological risk in agricultural watershed of the Three Gorges Reservoir region, a typical watershed, Wangjiagou, located in Fuling, where is in interior zones of the Three Gorges Reservoir region, was selected as the study object. Meanwhile, ArcGIS geo-statistics module was conducted for investigation of the Hg contents and distribution characteristics in soils of different land use types including dry land, farmland, woodland and settlements. Also the corresponding Hg pollution level and ecological risk were assessed. The results suggested that soil Hg contents in this watershed ranged from 9.47 to 94.57 microg x kg(-1), and the mean value was (34.23 +/- 16.23) microg x kg(-1). Higher Hg contents in surfaces of soils were observed in woodland, followed by farmland and settlement. The lowest was found in dry land. Surfaces of soils significantly showed Hg accumulation, and an obvious inverse correlation between soil Hg contents and soil depths was also observed in this study. Additionally, geo-statistics analysis showed a weak spatial correlation of soil Hg contents in this watershed, indicating the spatial distribution of soil Hg in this watershed was mainly influenced by several natural factors such as atmospheric wet-dry deposit, vegetation coverage and topography, instead of anthropogenic interference. Overall confirmative soil Hg pollution was not found in this watershed, which showed a very low pollution index (-0.08), but a moderate potential ecological risk still existed (the ecological risk index was 57), of which woodland had the highest potential risk. The total capacity of Hg in this watershed was 25.39 kg, among which dry land accounted for 69%.

  9. Computer simulation of the active site of human serum cholinesterase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kefang Jiao; Song Li; Zhengzheng Lu

    1996-12-31

    The first 3D-structure of acetylchelinesterase from Torpedo California electric organ (T.AChE) was published by JL. Sussman in 1991. We have simulated 3D-structure of human serum cholinesterase (H.BuChE) and the active site of H.BuChE. It is discovered by experiment that the residue of H.BuChE is still active site after a part of H.BuChE is cut. For example, the part of 21KD + 20KD is active site of H.BuChE. The 20KD as it is. Studies on these peptides by Hemelogy indicate that two active peptides have same negative electrostatic potential maps diagram. These negative electrostatic areas attached by acetyl choline with positivemore » electrostatic potency. We predict that 147...236 peptide of AChE could be active site because it was as 20KD as with negative electrostatic potential maps. We look forward to proving from other ones.« less

  10. De novo active sites for resurrected Precambrian enzymes

    NASA Astrophysics Data System (ADS)

    Risso, Valeria A.; Martinez-Rodriguez, Sergio; Candel, Adela M.; Krüger, Dennis M.; Pantoja-Uceda, David; Ortega-Muñoz, Mariano; Santoyo-Gonzalez, Francisco; Gaucher, Eric A.; Kamerlin, Shina C. L.; Bruix, Marta; Gavira, Jose A.; Sanchez-Ruiz, Jose M.

    2017-07-01

    Protein engineering studies often suggest the emergence of completely new enzyme functionalities to be highly improbable. However, enzymes likely catalysed many different reactions already in the last universal common ancestor. Mechanisms for the emergence of completely new active sites must therefore either plausibly exist or at least have existed at the primordial protein stage. Here, we use resurrected Precambrian proteins as scaffolds for protein engineering and demonstrate that a new active site can be generated through a single hydrophobic-to-ionizable amino acid replacement that generates a partially buried group with perturbed physico-chemical properties. We provide experimental and computational evidence that conformational flexibility can assist the emergence and subsequent evolution of new active sites by improving substrate and transition-state binding, through the sampling of many potentially productive conformations. Our results suggest a mechanism for the emergence of primordial enzymes and highlight the potential of ancestral reconstruction as a tool for protein engineering.

  11. Sedimentary and hydrological studies of the Holocene palaeofloods in the Shanxi-Shaanxi Gorge of the middle Yellow River, China

    NASA Astrophysics Data System (ADS)

    Li, Xiaogang; Huang, Chun Chang; Pang, Jiangli; Zha, Xiaochun; Ma, Yugai

    2015-01-01

    Holocene slackwater deposits along the river channels were used to study the magnitude and frequency of the palaeofloods that occurred prior to gauged and historical data sets all over the world. Palaeohydrological investigations along the Shanxi-Shaanxi Gorge of the middle Yellow River, China, identified palaeoflood slackwater deposits (SWDs) at several sites along the cliffs bordering the river channel. The SWDs are intercalated within Holocene eolian loess-soil profiles and clastic slope deposits. The palaeoflood SWDs were differentiated from eolian loess and soil by the sedimentary criteria and analytical results including magnetic susceptibility and particle-size distribution, similar to the flood SWDs in 2012, which indicated that these well-sorted palaeoflood SWD beds were deposited from the suspended sediment load in floodwaters. They have recorded the extraordinary palaeoflood events which occurred between 3200 and 3000 a BP as dated by the optically stimulated luminescence method in combination with pedostratigraphic correlations with the previously studied Holocene pedo-stratigraphy in the Yellow River drainage basin. Manning slope-area calculations estimate the peak discharged for these palaeoflood events to range from 43,290 to 49,830 m3/s. The drainage area of the study site is 489,900 km2. It is 2.0-2.5 times the largest gauged flood (21,000 m3/s) that has ever occurred since 1934. These events also occurred on Yellow River tributaries, including the Weihe, Jinghe and Qishuihe Rivers. These flood events are therefore considered to be a regional expression of known climatic events in the northern hemisphere and demonstrate Holocene climatic instability. This study provides important data in understanding the interactions between regional hydro-climatic systems and global change in semiarid and subhumid regions.

  12. Western Pond Turtle Head-starting and Reintroduction; 2004-2005 Annual Report.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Van Leuven, Susan; Allen, Harriet; Slavin, Kate

    This report covers the results of the western pond turtle head-starting and reintroduction project for the period of October 2004-September 2005. Wild hatchling western pond turtles from the Columbia River Gorge were reared at the Woodland Park and Oregon Zoos in 2004 and 2005 as part of the recovery effort for this Washington State endangered species. The objective of the program is to reduce losses to introduced predators like bullfrogs and largemouth bass by raising the hatchlings to a size where they are too large to be eaten by most of these predators. Thirty-five turtles were placed at the Woodlandmore » Park Zoo and 53 at the Oregon Zoo. Of these, 77 head-started juvenile turtles were released at three sites in the Columbia Gorge in 2005. Four were held back to attain more growth in captivity. Eleven were released at the Klickitat ponds, 22 at the Klickitat lake, 39 at the Skamania site, and 5 at Pierce National Wildlife Refuge (NWR). This brought the total number of head-start turtles released since 1991 to 257 for the Klickitat ponds, 136 for the Klickitat lake, 206 for the Skamania pond complex, and 255 at Pierce NWR. In 2005, 34 females from the two Columbia Gorge populations were equipped with transmitters and monitored for nesting activity. Twenty-four nests were located and protected; these produced 90 hatchlings. The hatchlings were collected in September and transported to the Oregon and Woodland Park zoos for rearing in the head-start program. During the 2005 field season trapping effort, 486 western pond turtles were captured in the Columbia Gorge, including 430 previously head-started turtles. These recaptures, together with confirmed nesting by head-start females and visual resightings, indicate the program is succeeding in boosting juvenile recruitment to increase the populations. Records were also collected on 216 individual painted turtles captured in 2005 during trapping efforts at Pierce NWR, to gather baseline information on this native

  13. Molecular dynamics explorations of active site structure in designed and evolved enzymes.

    PubMed

    Osuna, Sílvia; Jiménez-Osés, Gonzalo; Noey, Elizabeth L; Houk, K N

    2015-04-21

    This Account describes the use of molecular dynamics (MD) simulations to reveal how mutations alter the structure and organization of enzyme active sites. As proposed by Pauling about 70 years ago and elaborated by many others since then, biocatalysis is efficient when functional groups in the active site of an enzyme are in optimal positions for transition state stabilization. Changes in mechanism and covalent interactions are often critical parts of enzyme catalysis. We describe our explorations of the dynamical preorganization of active sites using MD, studying the fluctuations between active and inactive conformations normally concealed to static crystallography. MD shows how the various arrangements of active site residues influence the free energy of the transition state and relates the populations of the catalytic conformational ensemble to the enzyme activity. This Account is organized around three case studies from our laboratory. We first describe the importance of dynamics in evaluating a series of computationally designed and experimentally evolved enzymes for the Kemp elimination, a popular subject in the enzyme design field. We find that the dynamics of the active site is influenced not only by the original sequence design and subsequent mutations but also by the nature of the ligand present in the active site. In the second example, we show how microsecond MD has been used to uncover the role of remote mutations in the active site dynamics and catalysis of a transesterase, LovD. This enzyme was evolved by Tang at UCLA and Codexis, Inc., and is a useful commercial catalyst for the production of the drug simvastatin. X-ray analysis of inactive and active mutants did not reveal differences in the active sites, but relatively long time scale MD in solution showed that the active site of the wild-type enzyme preorganizes only upon binding of the acyl carrier protein (ACP) that delivers the natural acyl group to the active site. In the absence of bound ACP

  14. Relocating the Active-Site Lysine in Rhodopsin: 2. Evolutionary Intermediates.

    PubMed

    Devine, Erin L; Theobald, Douglas L; Oprian, Daniel D

    2016-08-30

    The visual pigment rhodopsin is a G protein-coupled receptor that covalently binds its retinal chromophore via a Schiff base linkage to an active-site Lys residue in the seventh transmembrane helix. Although this residue is strictly conserved among all type II retinylidene proteins, we found previously that the active-site Lys in bovine rhodopsin (Lys296) can be moved to three other locations (G90K, T94K, S186K) while retaining the ability to form a pigment with retinal and to activate transducin in a light-dependent manner [ Devine et al. ( 2013 ) Proc. Natl. Acad. Sci. USA 110 , 13351 - 13355 ]. Because the active-site Lys is not functionally constrained to be in helix seven, it is possible that it could relocate within the protein, most likely via an evolutionary intermediate with two active-site Lys. Therefore, in this study we characterized potential evolutionary intermediates with two Lys in the active site. Four mutant rhodopsins were prepared in which the original Lys296 was left untouched and a second Lys residue was substituted for G90K, T94K, S186K, or F293K. All four constructs covalently bind 11-cis-retinal, form a pigment, and activate transducin in a light-dependent manner. These results demonstrate that rhodopsin can tolerate a second Lys in the retinal binding pocket and suggest that an evolutionary intermediate with two Lys could allow migration of the Schiff base Lys to a position other than the observed, highly conserved location in the seventh TM helix. From sequence-based searches, we identified two groups of natural opsins, insect UV cones and neuropsins, that contain Lys residues at two positions in their active sites and also have intriguing spectral similarities to the mutant rhodopsins studied here.

  15. Geomorphological map of glaciated gorges in a granitic massif (Gredos range, Central Spain).

    NASA Astrophysics Data System (ADS)

    Campos, Néstor; Tanarro, Luis Miguel; Palacios, David

    2017-04-01

    A detailed geomorphological map on a 1:10,000 scale is presented for a high mountain area in Gredos range (Iberian Central System), this area is located in a granitic massif 160 km West of Madrid and comprises three gorges : La Vega, Taheña-Honda and La Nava. Only few detailed geomorphological maps of the Gredos range are available despite the wide diversity of landforms, in order to improve the understanding of this zone, this geomorphological map of the area has been produced, showing in detail the geomorphologic diversity of these gorges. The map was created with the aid of 25 cm resolution aerial photographs, 25 cm resolution satellite images, Iberpix 3D images provided by the Spanish National Geographic Institute and verified with field work. The landforms were delimitated with a stereoscope and satellite image pairs and digitized using GIS and CAD software, in some areas 3D glasses has been used with 3D images and the software Esri ArcScene. The landforms resulting from interpretation of aerial photographs and satellite images were classified using the IGUL (Institute of Geography, University of Lausanne) legend system (developed at the end of the 1980s) combined with the legend proposed by Peña et al. (1997) and some personal adaptations. The map legend includes 45 landforms divided into seven sections: structural, hydrography, fluvial, gravitative, glacial, nival and anthropic landforms. The use of both legend systems allows us to represent the landform types distributed over an area of 40 km2 and to identify the geomorphic processes involved in their morphogenesis, this variety of processes and landforms identified demonstrated that geomorphological cartography obtained by combining traditional image interpretation and GIS technology facilitates the production of geomorphological maps and the obtaining of valuable data for identify and understand surface processes and landforms. References: Maillard, B., Lambiel, C., Martin, S., Pellitero, R., Reynard, E

  16. Allosteric site-mediated active site inhibition of PBP2a using Quercetin 3-O-rutinoside and its combination.

    PubMed

    Rani, Nidhi; Vijayakumar, Saravanan; P T V, Lakshmi; Arunachalam, Annamalai

    2016-08-01

    Recent crystallographic study revealed the involvement of allosteric site in active site inhibition of penicillin binding protein (PBP2a), where one molecule of Ceftaroline (Cef) binds to the allosteric site of PBP2a and paved way for the other molecule (Cef) to bind at the active site. Though Cef has the potency to inhibit the PBP2a, its adverse side effects are of major concern. Previous studies have reported the antibacterial property of Quercetin derivatives, a group of natural compounds. Hence, the present study aims to evaluate the effect of Quercetin 3-o-rutinoside (Rut) in allosteric site-mediated active site inhibition of PBP2a. The molecular docking studies between allosteric site and ligands (Rut, Que, and Cef) revealed a better binding efficiency (G-score) of Rut (-7.790318) and Cef (-6.194946) with respect to Que (-5.079284). Molecular dynamic (MD) simulation studies showed significant changes at the active site in the presence of ligands (Rut and Cef) at allosteric site. Four different combinations of Rut and Cef were docked and their G-scores ranged between -6.320 and -8.623. MD studies revealed the stability of the key residue (Ser403) with Rut being at both sites, compared to other complexes. Morphological analysis through electron microscopy confirmed that combination of Rut and Cefixime was able to disturb the bacterial cell membrane in a similar fashion to that of Rut and Cefixime alone. The results of this study indicate that the affinity of Rut at both sites were equally good, with further validations Rut could be considered as an alternative for inhibiting MRSA growth.

  17. Numerical simulations of the impact of hydraulic anisotropy and anthropogenic activities on the complex hydrogeological system of the Lower Yarmouk Gorge

    NASA Astrophysics Data System (ADS)

    Tzoufka, Kalliopi; Siebert, Christian; Rödiger, Tino; Inbar, Nimrod; Shalev, Eyal; Möller, Peter; Raggad, Marwan; Rosenthal, Eliahu; Magri, Fabien

    2017-04-01

    Transient numerical simulations of coupled fluid flow and heat transport processes were conducted, to investigate (i) the impact of hydraulic anisotropy on a complex hydrogeological system and (ii) the anomalous geothermal gradient in the Upper Cretaceous (A7/B2) aquifer of the Lower Yarmouk Gorge (LYG). The N-S directed geological profile starts on the basalt-covered Golan Heights, crosses the LYG and ends at the Jordanian Ajloun Plateau. Heated fresh groundwaters ascend within the LYG from the confined A7/B2 limestone aquifer through artesian Meizar wells and artesian Muhkeibeh well field. A hydrogeological study based on high frequency water-table measurements at the Meizar wells suggested strong impact of abstraction at the Mukheibeh wells on the hydraulic head distribution north of the LYG (Shalev et al., 2015). Contrastingly, hydrochemical investigations conducted in the area concluded that recharge areas of the A7/B2 aquifer are: (i) the foothills of Mountain Hermon, (ii) the Ajloun Plateau and (iii) the Syrian Hauran Plateau, indicating the presence of a zone of high-hydraulic anisotropy along the main LYG axis. Due to this still debatable hydraulic feature, flow along the LYG principle axis is enhanced whilst flow perpendicular to it is constrained (Siebert et al., 2014). In agreement, transient simulations based on a NW-SE profile supported further the hypothesis of a structural feature existent (Magri et al., 2015). The modeled profile cross-cuts the heterogeneous zone and hence the heterogeneity was implemented as an impermeable zone by employing the Equivalent Porous Media approach. Initial 2D models managed to successfully reproduce the natural hydraulic head and temperature distributions. In subsequent simulations, by implementing Meizar abstraction rates, results revealed that mixed convection explains the anomalous temperature gradient in the area as temperature patterns of these simulations are in accordance with a temperature-depth borehole

  18. Assessing the influence of water level on schistosomiasis in Dongting Lake region before and after the construction of Three Gorges Dam.

    PubMed

    Li, Zhongwu; Nie, Xiaodong; Zhang, Yan; Huang, Jinquan; Huang, Bin; Zeng, Guangming

    2016-01-01

    Schistosomiasis is a severe public health problem in the Dongting Lake region, and its distribution, prevalence, and intensity of infection are particularly sensitive to environmental changes. In this study, the human and bovine schistosomiasis variations in the Dongting Lake region were studied from 1996 to 2010, and the relationships between schistosomiasis and water level were examined. Furthermore, based on these results, the potential effects of the Three Gorges Dam (TGD) on schistosomiasis were investigated. Results showed an increase in human schistosomiasis and in the scope of seriously affected regions, along with a decrease in bovine schistosomiasis. Human schistosomiasis was negatively correlated with water level during wet season (from May to October), particularly the average water level in October. This finding indicated that the decreasing water level may be highly related to the increasing of human schistosomiasis in the Dongting Lake region. Based on this result and the variation of schistosomiasis before and after the construction and operation of TGD, the impoundment of the Three Gorges reservoir is believed to decrease the water level and increase the contact between people and schistosomiasis. Therefore, the TGD, which is operated by regulating water and scheduling water operations, is not good for the control of human schistosomiasis in the Dongting Lake region. Although the extent of the influence of the TGD on schistosomiasis remains unclear, the influence of the TGD on preventing and controlling schistosomiasis should not be ignored.

  19. Influence of active site location on catalytic activity in de novo-designed zinc metalloenzymes.

    PubMed

    Zastrow, Melissa L; Pecoraro, Vincent L

    2013-04-17

    While metalloprotein design has now yielded a number of successful metal-bound and even catalytically active constructs, the question of where to put a metal site along a linear, repetitive sequence has not been thoroughly addressed. Often several possibilities in a given sequence may exist that would appear equivalent but may in fact differ for metal affinity, substrate access, or protein dynamics. We present a systematic variation of active site location for a hydrolytically active ZnHis3O site contained within a de novo-designed three-stranded coiled coil. We find that the maximal rate, substrate access, and metal-binding affinity are dependent on the selected position, while catalytic efficiency for p-nitrophenyl acetate hydrolysis can be retained regardless of the location of the active site. This achievement demonstrates how efficient, tailor-made enzymes which control rate, pKa, substrate and solvent access (and selectivity), and metal-binding affinity may be realized. These findings may be applied to the more advanced de novo design of constructs containing secondary interactions, such as hydrogen-bonding channels. We are now confident that changes to location for accommodating such channels can be achieved without location-dependent loss of catalytic efficiency. These findings bring us closer to our ultimate goal of incorporating the secondary interactions we believe will be necessary in order to improve both active site properties and the catalytic efficiency to be competitive with the native enzyme, carbonic anhydrase.

  20. Active Sites Environmental Monitoring Program: Mid-FY 1991 report

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ashwood, T.L.; Wickliff, D.S.; Morrissey, C.M.

    1991-10-01

    This report summarizes the activities of the Active Sites Environmental Monitoring Program (ASEMP) from October 1990 through March 1991. The ASEMP was established in 1989 by Solid Waste Operations and the Environmental Sciences Division to provide early detection and performance monitoring at active low-level radioactive waste (LLW) disposal sites in Solid Waste Storage Area (SWSA) 6 and transuranic (TRU) waste storage sites in SWSA 5 as required by chapters II and III of US Department of Energy Order 5820.2A. Monitoring results continue to demonstrate the no LLW is being leached from the storage vaults on the tumulus pads. Loading ofmore » vaults on Tumulus II began during this reporting period and 115 vaults had been loaded by the end of March 1991.« less

  1. Binding free energy calculations to rationalize the interactions of huprines with acetylcholinesterase.

    PubMed

    Nascimento, Érica C M; Oliva, Mónica; Andrés, Juan

    2018-05-01

    In the present study, the binding free energy of a family of huprines with acetylcholinesterase (AChE) is calculated by means of the free energy perturbation method, based on hybrid quantum mechanics and molecular mechanics potentials. Binding free energy calculations and the analysis of the geometrical parameters highlight the importance of the stereochemistry of huprines in AChE inhibition. Binding isotope effects are calculated to unravel the interactions between ligands and the gorge of AChE. New chemical insights are provided to explain and rationalize the experimental results. A good correlation with the experimental data is found for a family of inhibitors with moderate differences in the enzyme affinity. The analysis of the geometrical parameters and interaction energy per residue reveals that Asp72, Glu199, and His440 contribute significantly to the network of interactions between active site residues, which stabilize the inhibitors in the gorge. It seems that a cooperative effect of the residues of the gorge determines the affinity of the enzyme for these inhibitors, where Asp72, Glu199, and His440 make a prominent contribution.

  2. Binding free energy calculations to rationalize the interactions of huprines with acetylcholinesterase

    NASA Astrophysics Data System (ADS)

    Nascimento, Érica C. M.; Oliva, Mónica; Andrés, Juan

    2018-03-01

    In the present study, the binding free energy of a family of huprines with acetylcholinesterase (AChE) is calculated by means of the free energy perturbation method, based on hybrid quantum mechanics and molecular mechanics potentials. Binding free energy calculations and the analysis of the geometrical parameters highlight the importance of the stereochemistry of huprines in AChE inhibition. Binding isotope effects are calculated to unravel the interactions between ligands and the gorge of AChE. New chemical insights are provided to explain and rationalize the experimental results. A good correlation with the experimental data is found for a family of inhibitors with moderate differences in the enzyme affinity. The analysis of the geometrical parameters and interaction energy per residue reveals that Asp72, Glu199, and His440 contribute significantly to the network of interactions between active site residues, which stabilize the inhibitors in the gorge. It seems that a cooperative effect of the residues of the gorge determines the affinity of the enzyme for these inhibitors, where Asp72, Glu199, and His440 make a prominent contribution.

  3. Binding free energy calculations to rationalize the interactions of huprines with acetylcholinesterase

    NASA Astrophysics Data System (ADS)

    Nascimento, Érica C. M.; Oliva, Mónica; Andrés, Juan

    2018-05-01

    In the present study, the binding free energy of a family of huprines with acetylcholinesterase (AChE) is calculated by means of the free energy perturbation method, based on hybrid quantum mechanics and molecular mechanics potentials. Binding free energy calculations and the analysis of the geometrical parameters highlight the importance of the stereochemistry of huprines in AChE inhibition. Binding isotope effects are calculated to unravel the interactions between ligands and the gorge of AChE. New chemical insights are provided to explain and rationalize the experimental results. A good correlation with the experimental data is found for a family of inhibitors with moderate differences in the enzyme affinity. The analysis of the geometrical parameters and interaction energy per residue reveals that Asp72, Glu199, and His440 contribute significantly to the network of interactions between active site residues, which stabilize the inhibitors in the gorge. It seems that a cooperative effect of the residues of the gorge determines the affinity of the enzyme for these inhibitors, where Asp72, Glu199, and His440 make a prominent contribution.

  4. Smoothed particle hydrodynamic modeling of volcanic debris flows: Application to Huiloac Gorge lahars (Popocatépetl volcano, Mexico)

    NASA Astrophysics Data System (ADS)

    Haddad, Bouchra; Palacios, David; Pastor, Manuel; Zamorano, José Juan

    2016-09-01

    Lahars are among the most catastrophic volcanic processes, and the ability to model them is central to mitigating their effects. Several lahars recently generated by the Popocatépetl volcano (Mexico) moved downstream through the Huiloac Gorge towards the village of Santiago Xalitzintla. The most dangerous was the 2001 lahar, in which the destructive power of the debris flow was maintained throughout the extent of the flow. Identifying the zone of hazard can be based either on numerical or empirical models, but a calibration and validation process is required to ensure hazard map quality. The Geoflow-SPH depth integrated numerical model used in this study to reproduce the 2001 lahar was derived from the velocity-pressure version of the Biot-Zienkiewicz model, and was discretized using the smoothed particle hydrodynamics (SPH) method. The results of the calibrated SPH model were validated by comparing the simulated deposit depth with the field depth measured at 16 cross sections distributed strategically along the gorge channel. Moreover, the dependency of the results on topographic mesh resolution, initial lahar mass shape and dimensions is also investigated. The results indicate that to accurately reproduce the 2001 lahar flow dynamics the channel topography needed to be discretized using a mesh having a minimum 5 m resolution, and an initial lahar mass shape that adopted the source area morphology. Field validation of the calibrated model showed that there was a satisfactory relationship between the simulated and field depths, the error being less than 20% for 11 of the 16 cross sections. This study demonstrates that the Geoflow-SPH model was able to accurately reproduce the lahar path and the extent of the flow, but also reproduced other parameters including flow velocity and deposit depth.

  5. Three-dimensional structure of homodimeric cholesterol esterase-ligand complex at 1.4 Å resolution

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pletnev, V.; Addlagatta, A.; Wawrzak, Z.

    2010-03-08

    The three-dimensional structure of a Candida cylindracea cholesterol esterase (ChE) homodimer (534 x 2 amino acids) in complex with a ligand of proposed formula C{sub 23}H{sub 48}O{sub 2} has been determined at 1.4 {angstrom} resolution in space group P1 using synchrotron low-temperature data. The structure refined to R = 0.136 and R{sub free} = 0.169 and has revealed new stereochemical details in addition to those detected for the apo- and holo-forms at 1.9 and 2.0 {angstrom} resolution, respectively [Ghosh et al. (1995), Structure, 3, 279-288]. The cholesterol esterase structure is a dimer with four spatially separated interfacial contact areas andmore » two symmetry-related pairs of openings to an internal intradimer cavity. Hydrophobic active-site gorges in each subunit face each other across a central interfacial cavity. The ChE subunits have carbohydrate chains attached to their Asn314 and Asn351 residues, with two ordered N-acetyl-D-glucosoamine moieties visible at each site. The side chains of 14 residues have two alternative conformations with occupancy values of 0.5 {+-} 0.2. For each subunit the electron density in the enzyme active-site gorge is well modeled by a C{sub 23}-chain fatty acid.« less

  6. Mass Movement Susceptibility Mapping Using Satellite Optical Imagery Compared With INSAR Monitoring: Zigui County, Three Gorges Region, China

    NASA Astrophysics Data System (ADS)

    Kincal, Cem; Singleton, Andrew; Liu, Peng; Li, Zhenhong; Drummond, Jane; Hoey, Trevor; Muller, Jan-Peter; Qu, Wei; Zeng, Qiming; Zhang, Jingfa; Du, Peijun

    2010-10-01

    Mass movements on steep slopes are a major hazard to communities and infrastructure in the Three Gorges region, China. Developing susceptibility maps of mass movements is therefore very important in both current and future land use planning. This study employed satellite optical imagery and an ASTER GDEM (15 m) to derive various parameters (namely geology; slope gradient; proximity to drainage networks and proximity to lineaments) in order to create a GIS-based map of mass movement susceptibility. This map was then evaluated using highly accurate deformation signals processed using the Persistent Scatterer (PS) InSAR technique. Areas of high susceptibility correspond well to points of high subsidence, which provides a strong support of our susceptibility map.

  7. All the catalytic active sites of MoS 2 for hydrogen evolution

    DOE PAGES

    Li, Guoqing; Zhang, Du; Qiao, Qiao; ...

    2016-11-29

    MoS 2 presents a promising low-cost catalyst for the hydrogen evolution reaction (HER), but the understanding about its active sites has remained limited. Here we present an unambiguous study of the catalytic activities of all possible reaction sites of MoS 2, including edge sites, sulfur vacancies, and grain boundaries. We demonstrate that, in addition to the well-known catalytically active edge sites, sulfur vacancies provide another major active site for the HER, while the catalytic activity of grain boundaries is much weaker. Here, the intrinsic turnover frequencies (Tafel slopes) of the edge sites, sulfur vacancies, and grain boundaries are estimated tomore » be 7.5 s –1 (65–75 mV/dec), 3.2 s –1 (65–85 mV/dec), and 0.1 s –1 (120–160 mV/dec), respectively. We also demonstrate that the catalytic activity of sulfur vacancies strongly depends on the density of the vacancies and the local crystalline structure in proximity to the vacancies. Unlike edge sites, whose catalytic activity linearly depends on the length, sulfur vacancies show optimal catalytic activities when the vacancy density is in the range of 7–10%, and the number of sulfur vacancies in high crystalline quality MoS 2 is higher than that in low crystalline quality MoS 2, which may be related with the proximity of different local crystalline structures to the vacancies.« less

  8. Machine-Learning Methods Enable Exhaustive Searches for Active Bimetallic Facets and Reveal Active Site Motifs for CO 2 Reduction

    DOE PAGES

    Ulissi, Zachary W.; Tang, Michael T.; Xiao, Jianping; ...

    2017-07-27

    Bimetallic catalysts are promising for the most difficult thermal and electrochemical reactions, but modeling the many diverse active sites on polycrystalline samples is an open challenge. Here, we present a general framework for addressing this complexity in a systematic and predictive fashion. Active sites for every stable low-index facet of a bimetallic crystal are enumerated and cataloged, yielding hundreds of possible active sites. The activity of these sites is explored in parallel using a neural-network-based surrogate model to share information between the many density functional theory (DFT) relaxations, resulting in activity estimates with an order of magnitude fewer explicit DFTmore » calculations. Sites with interesting activity were found and provide targets for follow-up calculations. This process was applied to the electrochemical reduction of CO 2 on nickel gallium bimetallics and indicated that most facets had similar activity to Ni surfaces, but a few exposed Ni sites with a very favorable on-top CO configuration. This motif emerged naturally from the predictive modeling and represents a class of intermetallic CO 2 reduction catalysts. These sites rationalize recent experimental reports of nickel gallium activity and why previous materials screens missed this exciting material. Most importantly these methods suggest that bimetallic catalysts will be discovered by studying facet reactivity and diversity of active sites more systematically.« less

  9. Machine-Learning Methods Enable Exhaustive Searches for Active Bimetallic Facets and Reveal Active Site Motifs for CO 2 Reduction

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ulissi, Zachary W.; Tang, Michael T.; Xiao, Jianping

    Bimetallic catalysts are promising for the most difficult thermal and electrochemical reactions, but modeling the many diverse active sites on polycrystalline samples is an open challenge. Here, we present a general framework for addressing this complexity in a systematic and predictive fashion. Active sites for every stable low-index facet of a bimetallic crystal are enumerated and cataloged, yielding hundreds of possible active sites. The activity of these sites is explored in parallel using a neural-network-based surrogate model to share information between the many density functional theory (DFT) relaxations, resulting in activity estimates with an order of magnitude fewer explicit DFTmore » calculations. Sites with interesting activity were found and provide targets for follow-up calculations. This process was applied to the electrochemical reduction of CO 2 on nickel gallium bimetallics and indicated that most facets had similar activity to Ni surfaces, but a few exposed Ni sites with a very favorable on-top CO configuration. This motif emerged naturally from the predictive modeling and represents a class of intermetallic CO 2 reduction catalysts. These sites rationalize recent experimental reports of nickel gallium activity and why previous materials screens missed this exciting material. Most importantly these methods suggest that bimetallic catalysts will be discovered by studying facet reactivity and diversity of active sites more systematically.« less

  10. Mutation at a strictly conserved, active site tyrosine in the copper amine oxidase leads to uncontrolled oxygenase activity.

    PubMed

    Chen, Zhi-Wei; Datta, Saumen; Dubois, Jennifer L; Klinman, Judith P; Mathews, F Scott

    2010-08-31

    The copper amine oxidases carry out two copper-dependent processes: production of their own redox-active cofactor (2,4,5-trihydroxyphenylalanine quinone, TPQ) and the subsequent oxidative deamination of substrate amines. Because the same active site pocket must facilitate both reactions, individual active site residues may serve multiple roles. We have examined the roles of a strictly conserved active site tyrosine Y305 in the copper amine oxidase from Hansenula polymorpha kinetically, spetroscopically (Dubois and Klinman (2006) Biochemistry 45, 3178), and, in the present work, structurally. While the Y305A enzyme is almost identical to the wild type, a novel, highly oxygenated species replaces TPQ in the Y305F active sites. This new structure not only provides the first direct detection of peroxy intermediates in cofactor biogenesis but also indicates the critical control of oxidation chemistry that can be conferred by a single active site residue.

  11. New evidence of Yangtze delta recession after closing of the Three Gorges Dam

    PubMed Central

    Luo, X. X.; Yang, S. L.; Wang, R. S.; Zhang, C. Y.; Li, P.

    2017-01-01

    Many deltas are likely undergoing net erosion because of rapid decreases in riverine sediment supply and rising global sea levels. However, detecting erosion in subaqueous deltas is usually difficult because of the lack of bathymetric data. In this study, by comparing bathymetric data between 1981 and 2012 and surficial sediment grain sizes from the Yangtze subaqueous delta front over the last three decades, we found severe erosion and significant sediment coarsening in recent years since the construction of Three Gorges Dam (TGD), the largest dam in the world. We attributed these morphological and sedimentary variations mainly to the human-induced drastic decline of river sediment discharge. Combined with previous studies based on bathymetric data from different areas of the same delta, we theorize that the Yangtze subaqueous delta is experiencing overall (net) erosion, although local accumulation was also noted. We expect that the Yangtze sediment discharge will further decrease in the near future because of construction of new dams and delta recession will continue to occur. PMID:28145520

  12. Functional Evolution of PLP-dependent Enzymes based on Active-Site Structural Similarities

    PubMed Central

    Catazaro, Jonathan; Caprez, Adam; Guru, Ashu; Swanson, David; Powers, Robert

    2014-01-01

    Families of distantly related proteins typically have very low sequence identity, which hinders evolutionary analysis and functional annotation. Slowly evolving features of proteins, such as an active site, are therefore valuable for annotating putative and distantly related proteins. To date, a complete evolutionary analysis of the functional relationship of an entire enzyme family based on active-site structural similarities has not yet been undertaken. Pyridoxal-5’-phosphate (PLP) dependent enzymes are primordial enzymes that diversified in the last universal ancestor. Using the Comparison of Protein Active Site Structures (CPASS) software and database, we show that the active site structures of PLP-dependent enzymes can be used to infer evolutionary relationships based on functional similarity. The enzymes successfully clustered together based on substrate specificity, function, and three-dimensional fold. This study demonstrates the value of using active site structures for functional evolutionary analysis and the effectiveness of CPASS. PMID:24920327

  13. Chapter 12: Daily Patterns of Marbled Murrelet Activity at Inland Sites

    Treesearch

    Nancy L. Naslund; Brian P. O’Donnell

    1995-01-01

    Patterns in the daily activity of Marbled Murrelets (Brachyramphus marmoratus) at inland sites has been studied throughout their range from California to Alaska. Murrelets are most active at inland sites around dawn, and to a lesser degree, at dusk. Throughout their range, peak levels of activity (detections) occur in the hour around dawn, but...

  14. An Active Site Water Network in the Plasminogen Activator Pla from Yersinia pestis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Eren, Elif; Murphy, Megan; Goguen, Jon

    2010-08-13

    The plasminogen activator Pla from Yersinia pestis is an outer membrane protease (omptin) that is important for the virulence of plague. Here, we present the high-resolution crystal structure of wild-type, enzymatically active Pla at 1.9 {angstrom}. The structure shows a water molecule located between active site residues D84 and H208, which likely corresponds to the nucleophilic water. A number of other water molecules are present in the active site, linking residues important for enzymatic activity. The R211 sidechain in loop L4 is close to the nucleophilic water and possibly involved in the stabilization of the oxyanion intermediate. Subtle conformational changesmore » of H208 result from the binding of lipopolysaccharide to the outside of the barrel, explaining the unusual dependence of omptins on lipopolysaccharide for activity. The Pla structure suggests a model for the interaction with plasminogen substrate and provides a more detailed understanding of the catalytic mechanism of omptin proteases.« less

  15. Anisotropic Covalency Contributions to Superexchange Pathways in Type One Copper Active Sites

    PubMed Central

    2015-01-01

    Type one (T1) Cu sites deliver electrons to catalytic Cu active sites: the mononuclear type two (T2) Cu site in nitrite reductases (NiRs) and the trinuclear Cu cluster in the multicopper oxidases (MCOs). The T1 Cu and the remote catalytic sites are connected via a Cys-His intramolecular electron-transfer (ET) bridge, which contains two potential ET pathways: P1 through the protein backbone and P2 through the H-bond between the Cys and the His. The high covalency of the T1 Cu–S(Cys) bond is shown here to activate the T1 Cu site for hole superexchange via occupied valence orbitals of the bridge. This covalency-activated electronic coupling (HDA) facilitates long-range ET through both pathways. These pathways can be selectively activated depending on the geometric and electronic structure of the T1 Cu site and thus the anisotropic covalency of the T1 Cu–S(Cys) bond. In NiRs, blue (π-type) T1 sites utilize P1 and green (σ-type) T1 sites utilize P2, with P2 being more efficient. Comparing the MCOs to NiRs, the second-sphere environment changes the conformation of the Cys-His pathway, which selectively activates HDA for superexchange by blue π sites for efficient turnover in catalysis. These studies show that a given protein bridge, here Cys-His, provides different superexchange pathways and electronic couplings depending on the anisotropic covalencies of the donor and acceptor metal sites. PMID:25310460

  16. Functional evolution of PLP-dependent enzymes based on active-site structural similarities.

    PubMed

    Catazaro, Jonathan; Caprez, Adam; Guru, Ashu; Swanson, David; Powers, Robert

    2014-10-01

    Families of distantly related proteins typically have very low sequence identity, which hinders evolutionary analysis and functional annotation. Slowly evolving features of proteins, such as an active site, are therefore valuable for annotating putative and distantly related proteins. To date, a complete evolutionary analysis of the functional relationship of an entire enzyme family based on active-site structural similarities has not yet been undertaken. Pyridoxal-5'-phosphate (PLP) dependent enzymes are primordial enzymes that diversified in the last universal ancestor. Using the comparison of protein active site structures (CPASS) software and database, we show that the active site structures of PLP-dependent enzymes can be used to infer evolutionary relationships based on functional similarity. The enzymes successfully clustered together based on substrate specificity, function, and three-dimensional-fold. This study demonstrates the value of using active site structures for functional evolutionary analysis and the effectiveness of CPASS. © 2014 Wiley Periodicals, Inc.

  17. Development of a bacteria-based index of biotic integrity (Ba-IBI) for assessing ecological health of the Three Gorges Reservoir in different operation periods.

    PubMed

    Li, Yi; Yang, Nan; Qian, Bao; Yang, Zhengjian; Liu, Defu; Niu, Lihua; Zhang, Wenlong

    2018-05-30

    It is urgently needed to quantitatively assess ecological health of the Three Gorges Reservoir (TGR) when considering its special environmental conditions and temporal variations caused by reservoir operation. This study developed a bacteria-based index of biotic integrity (Ba-IBI) based on sediment samples collected along the TGR in low water level period, impoundment period and sluicing period, respectively. Reference conditions were defined using 8 ecological variables describing the hydromorphology and anthropogenic disturbances around the sites. Five core metrics, including % Acidobacteria, % Gemmatimonadetes, % Geobacter, Methanotroph and Phototroph, were selected after the screening processes. The developed index could clearly discriminate reference and impaired conditions and exhibited significant relationship with environmental parameters according to the redundancy (p < 0.01) and multivariable linear regression analysis (R 2  = 0.76). By implementing Ba-IBI in the TGR, the ecological health of the sampling sites was defined as "Excellent" (25%), "Good" (50%) and "Fair" (25%) separately. The spatial variation of biotic integrity was closely associated with environmental and ecological changes, especially the increase of nutrient concentrations. This study revealed a significant tendency that the ecological health in the low water level and sluicing periods was better than that in the impoundment period, which could be attributed to the hydrodynamic changes due to water level fluctuation. This study provides a comprehensive understanding of ecological health of the TGR in different operation periods and the index offers a guideline for the reservoir regulation in the similar areas. Copyright © 2018. Published by Elsevier B.V.

  18. Active-site solvent replenishment observed during human carbonic anhydrase II catalysis.

    PubMed

    Kim, Jin Kyun; Lomelino, Carrie L; Avvaru, Balendu Sankara; Mahon, Brian P; McKenna, Robert; Park, SangYoun; Kim, Chae Un

    2018-01-01

    Human carbonic anhydrase II (hCA II) is a zinc metalloenzyme that catalyzes the reversible hydration/dehydration of CO 2 /HCO 3 - . Although hCA II has been extensively studied to investigate the proton-transfer process that occurs in the active site, its underlying mechanism is still not fully understood. Here, ultrahigh-resolution crystallographic structures of hCA II cryocooled under CO 2 pressures of 7.0 and 2.5 atm are presented. The structures reveal new intermediate solvent states of hCA II that provide crystallographic snapshots during the restoration of the proton-transfer water network in the active site. Specifically, a new intermediate water (W I ') is observed next to the previously observed intermediate water W I , and they are both stabilized by the five water molecules at the entrance to the active site (the entrance conduit). Based on these structures, a water network-restructuring mechanism is proposed, which takes place at the active site after the nucleophilic attack of OH - on CO 2 . This mechanism explains how the zinc-bound water (W Zn ) and W1 are replenished, which are directly responsible for the reconnection of the His64-mediated proton-transfer water network. This study provides the first 'physical' glimpse of how a water reservoir flows into the hCA II active site during its catalytic activity.

  19. Design, synthesis and evaluation of some N-methylenebenzenamine derivatives as selective acetylcholinesterase (AChE) inhibitor and antioxidant to enhance learning and memory.

    PubMed

    Shrivastava, Sushant K; Srivastava, Pavan; Upendra, T V R; Tripathi, Prabhash Nath; Sinha, Saurabh K

    2017-02-15

    Series of some 3,5-dimethoxy-N-methylenebenzenamine and 4-(methyleneamino)benzoic acid derivatives comprising of N-methylenebenzenamine nucleus were designed, synthesized, characterized, and assessed for their acetylcholinesterase (AChE), butyrylcholinesterase (BChE) inhibitory, and antioxidant activity thereby improving learning and memory in rats. The IC 50 values of all the compound along with standard were determined on AChE and BChE enzyme. The free radical scavenging activity was also assessed by in vitro DPPH (2,2-diphenyl-1-picryl-hydrazyl) and hydrogen peroxide radical scavenging assay. The selective inhibitions of all compounds were observed against AChE in comparison with standard donepezil. The enzyme kinetic study of the most active compound 4 indicated uncompetitive AChE inhibition. The docking studies of compound 4 exhibited the worthy interaction on active-site gorge residues Phe330 and Trp279 responsible for its high affinity towards AChE, whereas lacking of the BChE inhibition was observed due to a wider gorge binding site and absence of important aromatic amino acids interactions. The ex vivo study confirmed AChE inhibition abilities of compound 4 at brain site. Further, a considerable decrease in escape latency period of the compound was observed in comparison with standard donepezil through in vivo Spatial Reference Memory (SRM) and Spatial Working Memory (SWM) models which showed the cognition-enhancing potential of compound 4. The in vivo reduced glutathione (GSH) estimation on rat brain tissue homogenate was also performed to evaluate free radical scavenging activity substantiated the antioxidant activity in learning and memory. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Mechanism and Catalytic Site Atlas (M-CSA): a database of enzyme reaction mechanisms and active sites.

    PubMed

    Ribeiro, António J M; Holliday, Gemma L; Furnham, Nicholas; Tyzack, Jonathan D; Ferris, Katherine; Thornton, Janet M

    2018-01-04

    M-CSA (Mechanism and Catalytic Site Atlas) is a database of enzyme active sites and reaction mechanisms that can be accessed at www.ebi.ac.uk/thornton-srv/m-csa. Our objectives with M-CSA are to provide an open data resource for the community to browse known enzyme reaction mechanisms and catalytic sites, and to use the dataset to understand enzyme function and evolution. M-CSA results from the merging of two existing databases, MACiE (Mechanism, Annotation and Classification in Enzymes), a database of enzyme mechanisms, and CSA (Catalytic Site Atlas), a database of catalytic sites of enzymes. We are releasing M-CSA as a new website and underlying database architecture. At the moment, M-CSA contains 961 entries, 423 of these with detailed mechanism information, and 538 with information on the catalytic site residues only. In total, these cover 81% (195/241) of third level EC numbers with a PDB structure, and 30% (840/2793) of fourth level EC numbers with a PDB structure, out of 6028 in total. By searching for close homologues, we are able to extend M-CSA coverage of PDB and UniProtKB to 51 993 structures and to over five million sequences, respectively, of which about 40% and 30% have a conserved active site. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  1. [Spatial Distribution Characteristics of Different Species Mercury in Water Body of Changshou Lake in Three Gorges Reservoir Region].

    PubMed

    Bai, Wei-yang; Zhang, Cheng; Zhao, Zheng; Tang, Zhen-ya; Wang, Ding-yong

    2015-08-01

    An investigation on the concentrations and the spatial distribution characteristics of different species of mercury in the water body of Changshou Lake in Three Gorges Reservoir region was carried out based on the AreGIS statistics module. The results showed that the concentration of the total mercury in Changshou Lake surface water ranged from 0.50 to 3.78 ng x L(-1), with an average of 1.51 ng x L(-1); the concentration of the total MeHg (methylmercury) ranged from 0.10 to 0.75 ng x L(-1), with an average of 0.23 ng x L(-1). The nugget effect value of total mercury in surface water (50.65%), dissolved mercury (49.80%), particulate mercury (29.94%) and the activity mercury (26.95%) were moderate spatial autocorrelation. It indicated that the autocorrelation was impacted by the intrinsic properties of sediments (such as parent materials and rocks, geological mineral and terrain), and on the other hand it was also disturbed by the exogenous input factors (such as aquaculture, industrial activities, farming etc). The nugget effect value of dissolved methylmercury (DMeHg) in Changshou lake surface water (3.49%) was less than 25%, showing significant strong spatial autocorrelation. The distribution was mainly controlled by environmental factors in water. The proportion of total MeHg in total Hg in Changshou Lake water reached 30% which was the maximum ratio of the total MeHg to total Hg in freshwater lakes and rivers. It implied that mercury was easily methylated in the environment of Chanashou Lake.

  2. Submergence Tolerance and Germination Dynamics of Roegneria nutans Seeds in Water-Level Fluctuation Zones with Different Water Rhythms in the Three Gorges Reservoir

    PubMed Central

    Zeng, Bo; Pan, Xiaojiao; Su, Xiaolei

    2016-01-01

    The Three Gorges Dam features two water-level fluctuation zones (WLFZs): the preupland drawdown zone (PU-DZ) and the preriparian drawdown zone (PR-DZ). To investigate the vegetation potential of Roegneria nutans in WLFZs, we compared the submergence tolerance and germination dynamics in the natural riparian zone (NRZ), PU-DZ and PR-DZ. We found that the NRZ seeds maintained an 81.3% intactness rate and >91% germination rate. The final seed germination rate and germination dynamics were consistent with those of the controls. Meanwhile, the PU-DZ seeds submerged at 5 m, 10 m, 15 m, and 20 m exhibited intactness rates of 70.5%, 79.95%, 40.75%, and 39.87%, respectively, and >75% germination. Furthermore, the PR-DZ seeds exhibited intactness rates of 22.44%, 61.13%, 81.87%, and 15.36% at 5 m, 10 m, 15 m, and 17 m, respectively, and 80% germination. The germination rates of the intact seeds submerged >10 m were >80%. Finally, the intact seeds germinated quickly in all WLFZs. The high proportion of intact seeds, rapid germination capacity, and high germination rate permit R. nutans seeds to adapt to the complicated water rhythms of the PU-DZ and PR-DZ and indicate the potential for their use in vegetation restoration and recovery. Thus, perennial seeds can be used for vegetation restoration in the WLFZs of large reservoirs and in other regions with water rhythms similar to the Three Gorges Reservoir. PMID:27031104

  3. Active Site Gate Dynamics Modulate the Catalytic Activity of the Ubiquitination Enzyme E2-25K.

    PubMed

    Rout, Manoj K; Lee, Brian L; Lin, Aiyang; Xiao, Wei; Spyracopoulos, Leo

    2018-05-03

    The ubiquitin proteasome system (UPS) signals for degradation of proteins through attachment of K48-linked polyubiquitin chains, or alterations in protein-protein recognition through attachment of K63-linked chains. Target proteins are ubiquitinated in three sequential chemical steps by a three-component enzyme system. Ubiquitination, or E2 enzymes, catalyze the central step by facilitating reaction of a target protein lysine with the C-terminus of Ub that is attached to the active site cysteine of the E2 through a thioester bond. E2 reactivity is modulated by dynamics of an active site gate, whose central residue packs against the active site cysteine in a closed conformation. Interestingly, for the E2 Ubc13, which specifically catalyzes K63-linked ubiquitination, the central gate residue adopts an open conformation. We set out to determine if active site gate dynamics play a role in catalysis for E2-25K, which adopts the canonical, closed gate conformation, and which selectively synthesizes K48-linked ubiquitin chains. Gate dynamics were characterized using mutagenesis of key residues, combined with enzyme kinetics measurements, and main chain NMR relaxation. The experimental data were interpreted with all atom MD simulations. The data indicate that active site gate opening and closing rates for E2-25K are precisely balanced.

  4. Characteristics of Heavy Metals and Pb Isotopic Composition in Sediments Collected from the Tributaries in Three Gorges Reservoir, China

    PubMed Central

    Gao, Bo; Zhou, Huaidong; Huang, Yong; Wang, Yuchun; Gao, Jijun; Liu, Xiaobo

    2014-01-01

    The concentrations, distribution, accumulation, and potential ecological risk of heavy metals (Cr, Cu, Zn, Ni, As, Pb, Cd, and Hg) in sediments from the Three Gorges Reservoir (TGR) tributaries were determined and studied. Pb isotopic compositions in sediments were also measured to effectively identify the potential Pb sources. The results showed that the average concentrations of heavy metals in sediment of TGR tributaries were higher than the local background values of soils and sediments in China. The assessment by Geoaccumulation Index indicated that Cu, Ni, and Hg were at the “slightly polluted” level and Cd was ranked as the “moderately polluted” level in tributary sediments of TGR. The assessment by Potential Ecological Risk Index showed that Hg and Cd were the predominant elements in tributary sediments in TGR. The Pb isotopic ratios in sediments varied from 1.171 to 1.202 for 206Pb/207Pb and from 2.459 to 2.482 for 208Pb/207Pb in TGR. All Pb isotopic ratios in sediments were similar to those from coal combustion, lead ores (the mining activities and smelting process), and cement material, indicating that these anthropogenic inputs may be the main sources for Pb pollution in sediments of TGR tributaries. PMID:24624045

  5. SABER: A computational method for identifying active sites for new reactions

    PubMed Central

    Nosrati, Geoffrey R; Houk, K N

    2012-01-01

    A software suite, SABER (Selection of Active/Binding sites for Enzyme Redesign), has been developed for the analysis of atomic geometries in protein structures, using a geometric hashing algorithm (Barker and Thornton, Bioinformatics 2003;19:1644–1649). SABER is used to explore the Protein Data Bank (PDB) to locate proteins with a specific 3D arrangement of catalytic groups to identify active sites that might be redesigned to catalyze new reactions. As a proof-of-principle test, SABER was used to identify enzymes that have the same catalytic group arrangement present in o-succinyl benzoate synthase (OSBS). Among the highest-scoring scaffolds identified by the SABER search for enzymes with the same catalytic group arrangement as OSBS were l-Ala d/l-Glu epimerase (AEE) and muconate lactonizing enzyme II (MLE), both of which have been redesigned to become effective OSBS catalysts, demonstrated by experiments. Next, we used SABER to search for naturally existing active sites in the PDB with catalytic groups similar to those present in the designed Kemp elimination enzyme KE07. From over 2000 geometric matches to the KE07 active site, SABER identified 23 matches that corresponded to residues from known active sites. The best of these matches, with a 0.28 Å catalytic atom RMSD to KE07, was then redesigned to be compatible with the Kemp elimination using RosettaDesign. We also used SABER to search for potential Kemp eliminases using a theozyme predicted to provide a greater rate acceleration than the active site of KE07, and used Rosetta to create a design based on the proteins identified. PMID:22492397

  6. SABER: a computational method for identifying active sites for new reactions.

    PubMed

    Nosrati, Geoffrey R; Houk, K N

    2012-05-01

    A software suite, SABER (Selection of Active/Binding sites for Enzyme Redesign), has been developed for the analysis of atomic geometries in protein structures, using a geometric hashing algorithm (Barker and Thornton, Bioinformatics 2003;19:1644-1649). SABER is used to explore the Protein Data Bank (PDB) to locate proteins with a specific 3D arrangement of catalytic groups to identify active sites that might be redesigned to catalyze new reactions. As a proof-of-principle test, SABER was used to identify enzymes that have the same catalytic group arrangement present in o-succinyl benzoate synthase (OSBS). Among the highest-scoring scaffolds identified by the SABER search for enzymes with the same catalytic group arrangement as OSBS were L-Ala D/L-Glu epimerase (AEE) and muconate lactonizing enzyme II (MLE), both of which have been redesigned to become effective OSBS catalysts, demonstrated by experiments. Next, we used SABER to search for naturally existing active sites in the PDB with catalytic groups similar to those present in the designed Kemp elimination enzyme KE07. From over 2000 geometric matches to the KE07 active site, SABER identified 23 matches that corresponded to residues from known active sites. The best of these matches, with a 0.28 Å catalytic atom RMSD to KE07, was then redesigned to be compatible with the Kemp elimination using RosettaDesign. We also used SABER to search for potential Kemp eliminases using a theozyme predicted to provide a greater rate acceleration than the active site of KE07, and used Rosetta to create a design based on the proteins identified. Copyright © 2012 The Protein Society.

  7. 'Unconventional' coordination chemistry by metal chelating fragments in a metalloprotein active site.

    PubMed

    Martin, David P; Blachly, Patrick G; Marts, Amy R; Woodruff, Tessa M; de Oliveira, César A F; McCammon, J Andrew; Tierney, David L; Cohen, Seth M

    2014-04-09

    The binding of three closely related chelators: 5-hydroxy-2-methyl-4H-pyran-4-thione (allothiomaltol, ATM), 3-hydroxy-2-methyl-4H-pyran-4-thione (thiomaltol, TM), and 3-hydroxy-4H-pyran-4-thione (thiopyromeconic acid, TPMA) to the active site of human carbonic anhydrase II (hCAII) has been investigated. Two of these ligands display a monodentate mode of coordination to the active site Zn(2+) ion in hCAII that is not recapitulated in model complexes of the enzyme active site. This unprecedented binding mode in the hCAII-thiomaltol complex has been characterized by both X-ray crystallography and X-ray spectroscopy. In addition, the steric restrictions of the active site force the ligands into a 'flattened' mode of coordination compared with inorganic model complexes. This change in geometry has been shown by density functional computations to significantly decrease the strength of the metal-ligand binding. Collectively, these data demonstrate that the mode of binding by small metal-binding groups can be significantly influenced by the protein active site. Diminishing the strength of the metal-ligand bond results in unconventional modes of metal coordination not found in typical coordination compounds or even carefully engineered active site models, and understanding these effects is critical to the rational design of inhibitors that target clinically relevant metalloproteins.

  8. Enhanced enzyme kinetic stability by increasing rigidity within the active site.

    PubMed

    Xie, Yuan; An, Jiao; Yang, Guangyu; Wu, Geng; Zhang, Yong; Cui, Li; Feng, Yan

    2014-03-14

    Enzyme stability is an important issue for protein engineers. Understanding how rigidity in the active site affects protein kinetic stability will provide new insight into enzyme stabilization. In this study, we demonstrated enhanced kinetic stability of Candida antarctica lipase B (CalB) by mutating the structurally flexible residues within the active site. Six residues within 10 Å of the catalytic Ser(105) residue with a high B factor were selected for iterative saturation mutagenesis. After screening 2200 colonies, we obtained the D223G/L278M mutant, which exhibited a 13-fold increase in half-life at 48 °C and a 12 °C higher T50(15), the temperature at which enzyme activity is reduced to 50% after a 15-min heat treatment. Further characterization showed that global unfolding resistance against both thermal and chemical denaturation also improved. Analysis of the crystal structures of wild-type CalB and the D223G/L278M mutant revealed that the latter formed an extra main chain hydrogen bond network with seven structurally coupled residues within the flexible α10 helix that are primarily involved in forming the active site. Further investigation of the relative B factor profile and molecular dynamics simulation confirmed that the enhanced rigidity decreased fluctuation of the active site residues at high temperature. These results indicate that enhancing the rigidity of the flexible segment within the active site may provide an efficient method for improving enzyme kinetic stability.

  9. GASS-WEB: a web server for identifying enzyme active sites based on genetic algorithms.

    PubMed

    Moraes, João P A; Pappa, Gisele L; Pires, Douglas E V; Izidoro, Sandro C

    2017-07-03

    Enzyme active sites are important and conserved functional regions of proteins whose identification can be an invaluable step toward protein function prediction. Most of the existing methods for this task are based on active site similarity and present limitations including performing only exact matches on template residues, template size restraints, despite not being capable of finding inter-domain active sites. To fill this gap, we proposed GASS-WEB, a user-friendly web server that uses GASS (Genetic Active Site Search), a method based on an evolutionary algorithm to search for similar active sites in proteins. GASS-WEB can be used under two different scenarios: (i) given a protein of interest, to match a set of specific active site templates; or (ii) given an active site template, looking for it in a database of protein structures. The method has shown to be very effective on a range of experiments and was able to correctly identify >90% of the catalogued active sites from the Catalytic Site Atlas. It also managed to achieve a Matthew correlation coefficient of 0.63 using the Critical Assessment of protein Structure Prediction (CASP 10) dataset. In our analysis, GASS was ranking fourth among 18 methods. GASS-WEB is freely available at http://gass.unifei.edu.br/. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  10. Facebook, Twitter Activities Sites, Location and Students' Interest in Learning

    ERIC Educational Resources Information Center

    Igbo, J. N.; Ezenwaji, Ifeyinwa; Ajuziogu, Christiana U.

    2018-01-01

    This study was carried out to ascertain the influence of social networking sites activities (twitter and Facebook) on secondary school students' interest in learning It also considered the impact of these social networking sites activities on location of the students. Two research questions and two null hypotheses guided the study. Mean and…

  11. Active Site Hydrophobicity and the Convergent Evolution of Paraoxonase Activity in Structurally Divergent Enzymes: The Case of Serum Paraoxonase 1

    PubMed Central

    2016-01-01

    Serum paraoxonase 1 (PON1) is a native lactonase capable of promiscuously hydrolyzing a broad range of substrates, including organophosphates, esters, and carbonates. Structurally, PON1 is a six-bladed β-propeller with a flexible loop (residues 70–81) covering the active site. This loop contains a functionally critical Tyr at position 71. We have performed detailed experimental and computational analyses of the role of selected Y71 variants in the active site stability and catalytic activity in order to probe the role of Y71 in PON1’s lactonase and organophosphatase activities. We demonstrate that the impact of Y71 substitutions on PON1’s lactonase activity is minimal, whereas the kcat for the paraoxonase activity is negatively perturbed by up to 100-fold, suggesting greater mutational robustness of the native activity. Additionally, while these substitutions modulate PON1’s active site shape, volume, and loop flexibility, their largest effect is in altering the solvent accessibility of the active site by expanding the active site volume, allowing additional water molecules to enter. This effect is markedly more pronounced in the organophosphatase activity than the lactonase activity. Finally, a detailed comparison of PON1 to other organophosphatases demonstrates that either a similar “gating loop” or a highly buried solvent-excluding active site is a common feature of these enzymes. We therefore posit that modulating the active site hydrophobicity is a key element in facilitating the evolution of organophosphatase activity. This provides a concrete feature that can be utilized in the rational design of next-generation organophosphate hydrolases that are capable of selecting a specific reaction from a pool of viable substrates. PMID:28026940

  12. Active Site Hydrophobicity and the Convergent Evolution of Paraoxonase Activity in Structurally Divergent Enzymes: The Case of Serum Paraoxonase 1.

    PubMed

    Blaha-Nelson, David; Krüger, Dennis M; Szeler, Klaudia; Ben-David, Moshe; Kamerlin, Shina Caroline Lynn

    2017-01-25

    Serum paraoxonase 1 (PON1) is a native lactonase capable of promiscuously hydrolyzing a broad range of substrates, including organophosphates, esters, and carbonates. Structurally, PON1 is a six-bladed β-propeller with a flexible loop (residues 70-81) covering the active site. This loop contains a functionally critical Tyr at position 71. We have performed detailed experimental and computational analyses of the role of selected Y71 variants in the active site stability and catalytic activity in order to probe the role of Y71 in PON1's lactonase and organophosphatase activities. We demonstrate that the impact of Y71 substitutions on PON1's lactonase activity is minimal, whereas the k cat for the paraoxonase activity is negatively perturbed by up to 100-fold, suggesting greater mutational robustness of the native activity. Additionally, while these substitutions modulate PON1's active site shape, volume, and loop flexibility, their largest effect is in altering the solvent accessibility of the active site by expanding the active site volume, allowing additional water molecules to enter. This effect is markedly more pronounced in the organophosphatase activity than the lactonase activity. Finally, a detailed comparison of PON1 to other organophosphatases demonstrates that either a similar "gating loop" or a highly buried solvent-excluding active site is a common feature of these enzymes. We therefore posit that modulating the active site hydrophobicity is a key element in facilitating the evolution of organophosphatase activity. This provides a concrete feature that can be utilized in the rational design of next-generation organophosphate hydrolases that are capable of selecting a specific reaction from a pool of viable substrates.

  13. Characterization of the active site properties of CYP4F12.

    PubMed

    Eksterowicz, John; Rock, Dan A; Rock, Brooke M; Wienkers, Larry C; Foti, Robert S

    2014-10-01

    Cytochrome P450 4F12 is a drug-metabolizing enzyme that is primarily expressed in the liver, kidney, colon, small intestine, and heart. The properties of CYP4F12 that may impart an increased catalytic selectivity (decreased promiscuity) were explored through in vitro metabolite elucidation, kinetic isotope effect experiments, and computational modeling of the CYP4F12 active site. By using astemizole as a probe substrate for CYP4F12 and CYP3A4, it was observed that although CYP4F12 favored astemizole O-demethylation as the primary route of metabolism, CYP3A4 was capable of metabolizing astemizole at multiple sites on the molecule. Deuteration of astemizole at the site of O-demethylation resulted in an isotope effect of 7.1 as well as an 8.3-fold decrease in the rate of clearance for astemizole by CYP4F12. Conversely, although an isotope effect of 3.8 was observed for the formation of the O-desmethyl metabolite when deuterated astemizole was metabolized by CYP3A4, there was no decrease in the clearance of astemizole. Development of a homology model of CYP4F12 based on the crystal structure of cytochrome P450 BM3 predicted an active site volume for CYP4F12 that was approximately 76% of the active site volume of CYP3A4. As predicted, multiple favorable binding orientations were available for astemizole docked into the active site of CYP3A4, but only a single binding orientation with the site of O-demethylation oriented toward the heme was identified for CYP4F12. Overall, it appears that although CYP4F12 may be capable of binding similar ligands to other cytochrome P450 enzymes such as CYP3A4, the ability to achieve catalytically favorable orientations may be inherently more difficult because of the increased steric constraints of the CYP4F12 active site. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

  14. Synthesis, characterization, X-ray crystallography, acetyl cholinesterase inhibition and antioxidant activities of some novel ketone derivatives of gallic hydrazide-derived Schiff bases.

    PubMed

    Gwaram, Nura Suleiman; Ali, Hapipah Mohd; Abdulla, Mahmood Ameen; Buckle, Michael J C; Sukumaran, Sri Devi; Chung, Lip Yong; Othman, Rozana; Alhadi, Abeer A; Yehye, Wageeh A; Hadi, A Hamid A; Hassandarvish, Pouya; Khaledi, Hamid; Abdelwahab, Siddig Ibrahim

    2012-02-28

    Alzheimer's disease (AD) is the most common form of dementia among older people and the pathogenesis of this disease is associated with oxidative stress. Acetylcholinesterase inhibitors with antioxidant activities are considered potential treatments for AD. Some novel ketone derivatives of gallic hydrazide-derived Schiff bases were synthesized and examined for their antioxidant activities and in vitro and in silico acetyl cholinesterase inhibition. The compounds were characterized using spectroscopy and X-ray crystallography. The ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays revealed that all the compounds have strong antioxidant activities. N-(1-(5-bromo-2-hydroxyphenyl)-ethylidene)-3,4,5-trihydroxybenzohydrazide (2) was the most potent inhibitor of human acetyl cholinesterase, giving an inhibition rate of 77% at 100 μM. Molecular docking simulation of the ligand-enzyme complex suggested that the ligand may be positioned in the enzyme's active-site gorge, interacting with residues in the peripheral anionic subsite (PAS) and acyl binding pocket (ABP). The current work warrants further preclinical studies to assess the potential for these novel compounds for the treatment of AD.

  15. Resilience in rural left-behind middle school students in Yunyang county of the Three Gorges area in China: a prospective cohort study.

    PubMed

    Luo, Yan; Wang, Hong; Lei, Xun; Guo, Xue; Huang, Ke; Liu, Qin

    2016-03-23

    The number of left-behind children in China is gradually increasing. This study aimed to understand the mental health status and changes in resilience of rural, left-behind middle school students in Yunyang County of the Three Gorges area in China. A prospective cohort study, including two follow-up surveys, with a frequency of once every 6 months was conducted among middle school students in Yunyang County. A self-designed questionnaire was used to obtain socio-demographic factors of participants, the Mental Health Test (MHT) scale was used to assess their mental health statuses, and resilience levels were collected using Resilience Youth Development Module (RYDM) scale at baseline and at the first and second follow-up investigations. Of the 1401 students who completed the baseline survey, 1322 students were eligible for the cohort study, of whom 1160 were investigated in the first follow-up survey. Ultimately, 1101 students completed the 1-year cohort. The detection rate of mental health problems for middle school students in the rural Three Gorges area was 5.64 %, and there was no significant difference between left-behind students (LBS) and non-left-behind students (NLBS) (χ (2) = 1.056, P = 0.304). The detection rates of medium resilience rose gradually (Z = 4.185, P = 0.000), while that of high resilience declined gradually (Z = 4.192, P = 0.000) in the baseline, first and second follow-up investigations. There was no significant difference between LBS and NLBS in resilience level (P > 0.05). The average RYDM scores were 2.990, 2.926, 2.904 among LBS in the baseline, first and second follow-up investigation, respectively, and the effect of time on the average RYDM scores was significant (F = 14.873, P = 0.000). The average MHT scores in LBS were 41.54, 39.79, 38.84 in the baseline, first and second follow-up investigations, respectively, and the detection rates of students who had psychological problems increased gradually (Z = 4.651, P = 0.000). The simple

  16. Deletion of loop fragment adjacent to active site diminishes the stability and activity of exo-inulinase.

    PubMed

    Arjomand, Maryam Rezaei; Habibi-Rezaei, Mehran; Ahmadian, Gholamreza; Hassanzadeh, Malihe; Karkhane, Ali Asghar; Asadifar, Mandana; Amanlou, Massoud

    2016-11-01

    Inulinases are classified as hydrolases and widely used in the food and medical industries. Here, we report the deletion of a six-membered adjacent active site loop fragment ( 74 YGSDVT 79 sequence) from third Ω-loop of the exo-inulinase containing aspartate residue from Aspergillus niger to study its structural and functional importance. Site-directed mutagenesis was used to create the mutant of the exo-inulinase (Δ6SL). To investigate the stability of the region spanning this loop, MD simulations were performed 80ns for 20-85 residues. Molecular docking was performed to compare the interactions in the active sites of enzymes with fructose as a ligand. Accordingly, the functional thermostability of the exo-inulinase was significantly decreased upon loop fragment deletion. Evaluation of the kinetics parameters (V max , K m , k cat and, k cat /K m ) and activation energy (E a ) of the catalysis of enzymes indicated the importance of the deleted sequence on the catalytic performance of the enzyme. In conclusion, six-membered adjacent active site loop fragment not only plays a crucial role in the stability of the enzyme, but also it involves in the enzyme catalysis through lowering the activation energy of the catalysis and effective improving the catalytic performance. Copyright © 2016. Published by Elsevier B.V.

  17. Evaluation of the Three Gorges Dam project using multi-criteria analysis (MCA) based on a sustainable perspective

    NASA Astrophysics Data System (ADS)

    Han, Yue; Zheng, Wei; Guo, Junshan; Ma, Yihe; Ding, Junqi; Zhu, Lingkai; Che, Yongqiang; Zhang, Yanpeng

    2018-02-01

    Abstract . The Three Gorges dam of China is one of the largest and expensive hydropower projects of the world. The four main purposes of the project are flood control,energy production, improved navigation and fresh water supply. The dam project has been completed and running successfully with the potential benefits. However, this project is still a controversial issue among many environmentalists and socialists due to various impacts. This study focuses on the benefit and the impacts of the project, and also evaluates the performance of the project using multi-criteria analysis (MCA) approach from a sustainable perspective. Different sustainability criteria related with the dam project have been identified and used for the ranking and rating process. The final result of MCA comes with this scoring process and pairwise comparison, which evaluates the performance of the project considering different positive and negative aspects.

  18. The Crystal Structure of a Quercetin 2,3-Dioxygenase from Bacillus subtilis Suggests Modulation of Enzyme Activity by a Change in the Metal Ion at the Active Site(s)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gopal, B.; Madan, Lalima L.; Betz, Stephen F.

    2010-11-10

    Common structural motifs, such as the cupin domains, are found in enzymes performing different biochemical functions while retaining a similar active site configuration and structural scaffold. The soil bacterium Bacillus subtilis has 20 cupin genes (0.5% of the total genome) with up to 14% of its genes in the form of doublets, thus making it an attractive system for studying the effects of gene duplication. There are four bicupins in B. subtilis encoded by the genes yvrK, yoaN, yxaG, and ywfC. The gene products of yvrK and yoaN function as oxalate decarboxylases with a manganese ion at the active site(s),more » whereas YwfC is a bacitracin synthetase. Here we present the crystal structure of YxaG, a novel iron-containing quercetin 2,3-dioxygenase with one active site in each cupin domain. Yxag is a dimer, both in solution and in the crystal. The crystal structure shows that the coordination geometry of the Fe ion is different in the two active sites of YxaG. Replacement of the iron at the active site with other metal ions suggests modulation of enzymatic activity in accordance with the Irving-Williams observation on the stability of metal ion complexes. This observation, along with a comparison with the crystal structure of YvrK determined recently, has allowed for a detailed structure-function analysis of the active site, providing clues to the diversification of function in the bicupin family of proteins.« less

  19. [Three-dimensional Fluorescence Spectral Characteristics of Different Molecular Weight Fractionations of Dissolved Organic Matter in the Water-level Fluctuation Zones of Three Gorges Reservoir Areas].

    PubMed

    Chen, Xue-shuang; Jiang, Tao; Lu, Song; Wei, Shi-qiang; Wang, Ding-yong; Yan, Jin-long

    2016-03-15

    The study of the molecular weight (MW) fractions of dissolved organic matter (DOM) in aquatic environment is of interests because the size plays an important role in deciding the biogeochemical characteristics of DOM. Thus, using ultrafiltration ( UF) technique combined with three-dimensional fluorescence spectroscopy, DOM samples from four sampling sites in typical water-level fluctuation zones of Three Gorge Reservoir areas were selected to investigate the differences of properties and sources of different DOM MW fractions. The results showed that in these areas, the distribution of MW fractions was highly dispersive, but the approximately equal contributions from colloidal (Mr 1 x 10³-0.22 µm) and true dissolved fraction (Mr < 1 x 10³) to the total DOC concentration were found. Four fluorescence signals (humic-like A and C; protein-like B and T) were observed in all MW fractions including bulk DOM, which showed the same distribution trend: true dissolved > low MW (Mr 1 x 10³-10 x 10³) > medium MW (Mr 10 x 10³-30 x 10³) > high MW (Mr 30 x 10³-0.22 µm). Additionally, with decreasing MW fraction, fluorescence index (FI) and freshness index (BIX) increased suggesting enhanced signals of autochthonous inputs, whereas humification index ( HIX) decreased indicating lowe humification degree. It strongly suggested that terrestrial input mainly affected the composition and property of higher MW fractions of DOM, as compared to lower MW and true dissolved fractions that were controlled by autochthonous sources such as microbial and alga activities, instead of allochthonous sources. Meanwhile, the riparian different land-use types also affected obviously on the characteristics of DOM. Therefore, higher diversity of land-use types, and also higher complexity of ecosystem and landscapes, induced higher heterogeneity of fluorescence components in different MW fraction of DOM.

  20. A hybrid QM/MM simulation study of intramolecular proton transfer in the pyridoxal 5'-phosphate in the active site of transaminase: influence of active site interaction on proton transfer.

    PubMed

    Dutta Banik, Sindrila; Chandra, Amalendu

    2014-09-25

    Pyridoxal 5'-phosphate (PLP) Schiff base, a versatile cofactor, exhibits a tautomeric equilibrium that involves an intramolecular proton transfer between the N-protonated zwitterionic ketoenamine tautomer and the O-protonated covalent enolimine tautomer. It has been postulated that for the catalytic activity, the PLP has to be in the zwitterionic ketoenamine tautomeric form. However, the exact position of the tautomeric equilibrium of Schiff base in the active site of PLP-dependent enzyme is not known yet. In the present work, we investigated the tautomeric equilibrium for the external aldimine state of PLP aspartate (PLP-Asp) Schiff base in the active site of aspartate aminotransferase (AspAT) using combined quantum mechanical and molecular mechanical simulations. The main focus of the present study is to analyze the factors that control the tautomeric equilibrium in the active sites of various PLP-dependent enzymes. The results show that the ketoenamine tautomer is more preferred than the enolimine tautomer both in the gas and aqueous phases as well as in the active site of AspAT. Current simulations show that the active site of AspAT is more suitable for the ketoenamine tautomer compared to the enolimine tautomer. Interestingly, the Tyr225 acts as a proton donor to the phenolic oxygen in the ketoenamine tautomer, while in the covalent enolimine tautomer, it acts as a proton acceptor to the phenolic oxygen. Finally, the metadynamics study confirms this result. The calculated free energy barrier is about 7.5 kcal/mol. A comparative analysis of the microenvironment created by the active site residues of three different PLP-dependent enzymes (aspartate aminotransferase, Dopa decarboxylase, and Ala-racemase) has been carried out to understand the controlling factor(s) of the tautomeric equilibrium. The analysis shows that the intermolecular hydrogen bonding between active site residues and the phenolic oxygen of PLP shifts the tautomeric equilibrium toward the N

  1. Spatial and temporal variations in non-point source losses of nitrogen and phosphorus in a small agricultural catchment in the Three Gorges Region.

    PubMed

    Chen, Chenglong; Gao, Ming; Xie, Deti; Ni, Jiupai

    2016-04-01

    Losses of agricultural pollutants from small catchments are a major issue for water quality in the Three Gorges Region. Solutions are urgently needed. However, before pollutant losses can be controlled, information about spatial and temporal variations in pollutant losses is needed. The study was carried out in the Wangjiagou catchment, a small agricultural catchment in Fuling District, Chongqing, and the data about non-point source losses of nitrogen and phosphorus was collected here. Water samples were collected daily by an automatic water sampler at the outlets of two subcatchments from 2012 to 2014. Also, samples of surface runoff from 28 sampling sites distributed through the subcatchments were collected during 12 rainfall events in 2014. A range of water quality variables were analyzed for all samples and were used to demonstrate the variation in non-point losses of nitrogen and phosphorus over a range of temporal and spatial scales and in different types of rainfall in the catchment. Results showed that there was a significant linear correlation between the mass concentrations of total nitrogen (TN) and nitrate (NO3-N) in surface runoff and that the relationship was maintained with changes in time. Concentrations of TN and NO3-N peaked after fertilizer was applied to crops in spring and autumn; concentrations decreased rapidly after the peak values in spring but declined slowly in autumn. N and P concentrations fluctuated more and showed a greater degree of dispersion during the spring crop cultivation period than those in autumn. Concentrations of TN and NO3-N in surface runoff were significantly and positively correlated with the proportion of the area that was planted with corn and mustard tubers, but were negatively correlated with the proportion of the area taken up with rice and mulberry plantations. The average concentrations of TN and NO3-N in surface runoff reached the highest level from the sampling points at the bottom of the land used for corn

  2. Community Update on Site Activities, July 19, 2013

    EPA Pesticide Factsheets

    In an effort to engage and inform community members interested in the New Bedford Harbor Superfund Site cleanup, EPA will be issuing periodic topic-based fact sheets that will provide background information and updates about ongoing activities.

  3. Structural and Kinetic Analyses of Macrophage Migration Inhibitory Factor Active Site Interactions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Crichlow, G.; Lubetsky, J; Leng, L

    Macrophage migration inhibitory factor (MIF) is a secreted protein expressed in numerous cell types that counters the antiinflammatory effects of glucocorticoids and has been implicated in sepsis, cancer, and certain autoimmune diseases. Interestingly, the structure of MIF contains a catalytic site resembling the tautomerase/isomerase sites of microbial enzymes. While bona fide physiological substrates remain unknown, model substrates have been identified. Selected compounds that bind in the tautomerase active site also inhibit biological functions of MIF. It had previously been shown that the acetaminophen metabolite, N-acetyl-p-benzoquinone imine (NAPQI), covalently binds to the active site of MIF. In this study, kinetic datamore » indicate that NAPQI inhibits MIF both covalently and noncovalently. The structure of MIF cocrystallized with NAPQI reveals that the NAPQI has undergone a chemical alteration forming an acetaminophen dimer (bi-APAP) and binds noncovalently to MIF at the mouth of the active site. We also find that the commonly used protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), forms a covalent complex with MIF and inhibits the tautomerase activity. Crystallographic analysis reveals the formation of a stable, novel covalent bond for PMSF between the catalytic nitrogen of the N-terminal proline and the sulfur of PMSF with complete, well-defined electron density in all three active sites of the MIF homotrimer. Conclusions are drawn from the structures of these two MIF-inhibitor complexes regarding the design of novel compounds that may provide more potent reversible and irreversible inhibition of MIF.« less

  4. Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites.

    PubMed

    Chen, Baoyu; Chou, Hui-Ting; Brautigam, Chad A; Xing, Wenmin; Yang, Sheng; Henry, Lisa; Doolittle, Lynda K; Walz, Thomas; Rosen, Michael K

    2017-09-26

    The Rho GTPase Rac1 activates the WAVE regulatory complex (WRC) to drive Arp2/3 complex-mediated actin polymerization, which underpins diverse cellular processes. Here we report the structure of a WRC-Rac1 complex determined by cryo-electron microscopy. Surprisingly, Rac1 is not located at the binding site on the Sra1 subunit of the WRC previously identified by mutagenesis and biochemical data. Rather, it binds to a distinct, conserved site on the opposite end of Sra1. Biophysical and biochemical data on WRC mutants confirm that Rac1 binds to both sites, with the newly identified site having higher affinity and both sites required for WRC activation. Our data reveal that the WRC is activated by simultaneous engagement of two Rac1 molecules, suggesting a mechanism by which cells may sense the density of active Rac1 at membranes to precisely control actin assembly.

  5. Monocopper active site for partial methane oxidation in Cu-exchanged 8MR zeolites

    DOE PAGES

    Kulkarni, Ambarish R.; Zhao, Zhi -Jian; Siahrostami, Samira; ...

    2016-08-17

    Direct conversion of methane to methanol using oxygen is experiencing renewed interest owing to the availability of new natural gas resources. Copper-exchanged zeolites such as mordenite and ZSM-5 have shown encouraging results, and di- and tri-copper species have been suggested as active sites. Recently, small eight-membered ring (8MR) zeolites including SSZ-13, -16, and -39 have been shown to be active for methane oxidation, but the active sites and reaction mechanisms in these 8MR zeolites are not known. In this work, we use density functional theory (DFT) calculations to systematically evaluate monocopper species as active sites for the partial methane oxidationmore » reaction in Cu-exchanged SSZ-13. On the basis of kinetic and thermodynamic arguments, we suggest that [Cu IIOH] + species in the 8MR are responsible for the experimentally observed activity. Furthermore, our results successfully explain the available spectroscopic data and experimental observations including (i) the necessity of water for methanol extraction and (ii) the effect of Si/Al ratio on the catalyst activity. Monocopper species have not yet been suggested as an active site for the partial methane oxidation reaction, and our results suggest that [Cu IIOH] + active site may provide complementary routes for methane activation in zeolites in addition to the known [Cu–O–Cu] 2+ and Cu 3O 3 motifs.« less

  6. What Motivates Young Adults to Talk About Physical Activity on Social Network Sites?

    PubMed

    Zhang, Ni; Campo, Shelly; Yang, Jingzhen; Eckler, Petya; Snetselaar, Linda; Janz, Kathleen; Leary, Emily

    2017-06-22

    Electronic word-of-mouth on social network sites has been used successfully in marketing. In social marketing, electronic word-of-mouth about products as health behaviors has the potential to be more effective and reach more young adults than health education through traditional mass media. However, little is known about what motivates people to actively initiate electronic word-of-mouth about health behaviors on their personal pages or profiles on social network sites, thus potentially reaching all their contacts on those sites. This study filled the gap by applying a marketing theoretical model to explore the factors associated with electronic word-of-mouth on social network sites about leisure-time physical activity. A Web survey link was sent to undergraduate students at one of the Midwestern universities and 439 of them completed the survey. The average age of the 439 participants was 19 years (SD=1 year, range: 18-24). Results suggested that emotional engagement with leisure-time physical activity (ie, affective involvement in leisure-time physical activity) predicted providing relevant opinions or information on social network sites. Social network site users who perceived stronger ties with all their contacts were more likely to provide and seek leisure-time physical activity opinions and information. People who provided leisure-time physical activity opinions and information were more likely to seek opinions and information, and people who forwarded information about leisure-time physical activity were more likely to chat about it. This study shed light on the application of the electronic word-of-mouth theoretical framework in promoting health behaviors. The findings can also guide the development of future social marketing interventions using social network sites to promote leisure-time physical activity. ©Ni Zhang, Shelly Campo, Jingzhen Yang, Petya Eckler, Linda Snetselaar, Kathleen Janz, Emily Leary. Originally published in the Journal of Medical

  7. What Motivates Young Adults to Talk About Physical Activity on Social Network Sites?

    PubMed Central

    Campo, Shelly; Yang, Jingzhen; Eckler, Petya; Snetselaar, Linda; Janz, Kathleen; Leary, Emily

    2017-01-01

    Background Electronic word-of-mouth on social network sites has been used successfully in marketing. In social marketing, electronic word-of-mouth about products as health behaviors has the potential to be more effective and reach more young adults than health education through traditional mass media. However, little is known about what motivates people to actively initiate electronic word-of-mouth about health behaviors on their personal pages or profiles on social network sites, thus potentially reaching all their contacts on those sites. Objective This study filled the gap by applying a marketing theoretical model to explore the factors associated with electronic word-of-mouth on social network sites about leisure-time physical activity. Methods A Web survey link was sent to undergraduate students at one of the Midwestern universities and 439 of them completed the survey. Results The average age of the 439 participants was 19 years (SD=1 year, range: 18-24). Results suggested that emotional engagement with leisure-time physical activity (ie, affective involvement in leisure-time physical activity) predicted providing relevant opinions or information on social network sites. Social network site users who perceived stronger ties with all their contacts were more likely to provide and seek leisure-time physical activity opinions and information. People who provided leisure-time physical activity opinions and information were more likely to seek opinions and information, and people who forwarded information about leisure-time physical activity were more likely to chat about it. Conclusions This study shed light on the application of the electronic word-of-mouth theoretical framework in promoting health behaviors. The findings can also guide the development of future social marketing interventions using social network sites to promote leisure-time physical activity. PMID:28642215

  8. Landslides in the western Columbia Gorge, Skamania County, Washington

    USGS Publications Warehouse

    Pierson, Thomas C.; Evarts, Russell C.; Bard, Joseph A.

    2016-11-04

    SummaryRecent light detection and ranging (lidar) imagery has allowed us to identify and map a large number of previously unrecognized landslides, or slides, in heavily forested terrain in the western Columbia Gorge, Skamania County, Washington, and it has revealed that the few previously recognized areas of instability are actually composites of multiple smaller landslides. The high resolution of the imagery further reveals that landslides in the map area have complex movement histories and span a wide range of relative ages. Movement histories are inferred from relative landslide locations and crosscutting relations of surface features. Estimated age ranges are based on (1) limited absolute dating; (2) relative fineness of landscape surface textures, calibrated by comparison with surfaces of currently active and dated landslides as interpreted from interferometric synthetic aperture radar (InSAR), global positioning system (GPS), and historical records; (3) sharpness and steepness of larger-scale surface morphologic features, calibrated by comparison with similar dated features in other regions; (4) degree of surface erosion; and (5) evidence of erosion or deposition by late Pleistocene (15–22 ka) Missoula floods at or below 200 m altitude. The relative age categories are recent (0 to ~1,000 years old), intermediate-age (~1,000 to ~15,000 years old), and old (>~15,000 years old). Within the 221.5 km2 map area, we identified 215 discrete landslides, covering 140.9 km2 (64 percent of the map area). At least 12 of the recent landslides are currently moving or have moved within the last two decades. Mapping for this study expanded the area of previously recognized unstable terrain by 56 percent. Landslide geometries suggest that more than half (62 percent) of these slope failures are translational landslides or composite landslides with translational elements, with failure occurring along gently sloping bedding planes in zones of deeply weathered, locally clay rich

  9. Corrosion Research And Web Site Activities

    NASA Technical Reports Server (NTRS)

    Heidersbach, Robert H.

    2001-01-01

    This report covers corrosion-related activities at the NASA Kennedy Space Center during the summer of 2000. The NASA Kennedy Space Center's corrosion web site, corrosion.ksc.nasa.gov, was updated with new information based on feedback over the past two years. The methodology for a two-year atmospheric exposure testing program to study the effectiveness of commercial chemicals sold for rinsing aircraft and other equipment was developed and some preliminary laboratory chemical analyses are presented.

  10. Corrosion Research and Web Site Activities

    NASA Technical Reports Server (NTRS)

    Heidersbach, Robert H.

    2002-01-01

    This report covers corrosion-related activities at the NASA Kennedy Space Center during the summer of 2000. The NASA Kennedy Space Center's corrosion web site, corrosion.ksc.nasa.gov, was updated with new information based on feedback over the past two years. The methodology for a two-year atmospheric exposure testing program to study the effectiveness of commercial chemicals sold for rinsing aircraft and other equipment was developed and some preliminary laboratory chemical analyses are presented.

  11. Channeling by Proximity: The Catalytic Advantages of Active Site Colocalization Using Brownian Dynamics.

    PubMed

    Bauler, Patricia; Huber, Gary; Leyh, Thomas; McCammon, J Andrew

    2010-05-06

    Nature often colocalizes successive steps in a metabolic pathway. Such organization is predicted to increase the effective concentration of pathway intermediates near their recipient active sites and to enhance catalytic efficiency. Here, the pathway of a two-step reaction is modeled using a simple spherical approximation for the enzymes and substrate particles. Brownian dynamics are used to simulate the trajectory of a substrate particle as it diffuses between the active site zones of two different enzyme spheres. The results approximate distances for the most effective reaction pathways, indicating that the most effective reaction pathway is one in which the active sites are closely aligned. However, when the active sites are too close, the ability of the substrate to react with the first enzyme was hindered, suggesting that even the most efficient orientations can be improved for a system that is allowed to rotate or change orientation to optimize the likelihood of reaction at both sites.

  12. Cooperative activation of cardiac transcription through myocardin bridging of paired MEF2 sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Anderson, Courtney M.; Hu, Jianxin; Thomas, Reuben

    2017-03-28

    Enhancers frequently contain multiple binding sites for the same transcription factor. These homotypic binding sites often exhibit synergy, whereby the transcriptional output from two or more binding sites is greater than the sum of the contributions of the individual binding sites alone. Although this phenomenon is frequently observed, the mechanistic basis for homotypic binding site synergy is poorly understood. Here in this paper, we identify a bona fide cardiac-specific Prkaa2 enhancer that is synergistically activated by homotypic MEF2 binding sites. We show that two MEF2 sites in the enhancer function cooperatively due to bridging of the MEF2C-bound sites by themore » SAP domain-containing co-activator protein myocardin, and we show that paired sites buffer the enhancer from integration site-dependent effects on transcription in vivo. Paired MEF2 sites are prevalent in cardiac enhancers, suggesting that this might be a common mechanism underlying synergy in the control of cardiac gene expression in vivo.« less

  13. Preliminary Magnetostratigraphic Study of the Split Mountain and Lower Imperial Groups, Split Mountain Gorge, Western Salton Trough, CA

    NASA Astrophysics Data System (ADS)

    Fluette, A. L.; Housen, B. A.; Dorsey, R. J.

    2004-12-01

    We present preliminary results of a magnetostratigraphic study of Miocene-Pliocene sedimentary rocks of the Split Mt. and lower Imperial Groups exposed in Split Mt. Gorge and eastern Fish Creek-Vallecito basin, western Salton Trough. Precise age control for the base of this thick section is needed to improve our understanding of the early history of extension-related subsidence in this region. The geologic setting and stratigraphic framework are known from previous work by Dibblee (1954, 1996), Woodard (1963), Kerr (1982), Winker (1987), Kerr and Kidwell (1991), Winker and Kidwell (1986; 1996), and others. We have analyzed Upper Miocene to lower Pliocene strata exposed in a conformable section in Split Mt. Gorge, including (in order from the base; nomenclature of Winker and Kidwell, 1996): (1) Split Mt. Group: Red Rock Fm alluvial sandstone; Elephant Trees alluvial conglomerate; and lower megabreccia unit; and (2) lower part of Imperial Group, including: Fish Creek Gypsum; proximal to distal turbidites of the Latrania Fm and Wind Caves Mbr of Deguynos Fm; upper megabreccia unit; marine mudstone and rhythmites of the Mud Hills Mbr (Deguynos Fm); and the basal part of the Yuha Mbr (Deguynos Fm). Measured thickness from the base of the Elephant Trees Cgl to the base of the Yuha Mbr is about 1050 m, consistent with previous measurements of Winker (1987). Paleomagnetic samples were collected at approximately 10 m intervals throughout this section. The upper portion of our sampled section overlaps with the lower part of the section sampled for magnetostratigraphic study by Opdyke et al. (1977) and Johnson et al. (1983). They interpreted the base of their section to be about 4.3 Ma, and calculated an average sedimentation rate of approximately 5.5 mm/yr for the lower part of their section. Good-quality preliminary results from 15 paleomagnetic sites distributed throughout our sampled section permit preliminary identification of 6 polarity zones. Based on regional mapping

  14. Evaluation of the dynamic mobilization of vanadium in tributary sediments of the Three Gorges Reservoir after water impoundment

    NASA Astrophysics Data System (ADS)

    Gao, Bo; Gao, Li; Zhou, Yang; Xu, Dongyu; Zhao, Xingjuan

    2017-08-01

    The Three Gorges Reservoir (TGR) is the largest water resource protection zone in China, and environmental safety is crucial to its operation. For both aqueous and sediment phases, diffusive gradients in thin films (DGT), total vanadium (V) concentration (CTotal-V), and community Bureau of Reference (BCR) sequential extraction data were used to measure the pollution characteristics, horizontal and vertical distributions of DGT-labile V, and the dynamic mobilization of V in a typical tributary (the Meixi River) of the TGR. The results showed that CTotal-V in the surface sediments were obviously higher than the background values in sediment and soil, indicating a potential anthropogenic input of V in this area. A positive relationship was found between total organic carbon (TOC) and CTotal-V in the sediments, indicating that the pollution characteristics of V were associated with TOC. In addition, horizontal and vertical distributions of the fluxes of DGT-labile V (FDGT-V) varied among the four DGT probes. In the same DGT probe, the horizontal distributions (0-6 mm, 6-12 mm and 12-18 mm) of FDGT-V were similar in the overlying water; however, the values showed a poor coincidence with those recorded in the sediment. The vertical distribution of FDGT-V in the same DGT probe showed similar tendencies. In fact, CDGT-V is significantly negatively correlated with CDGT-Fe, demonstrating that V had an inversely diffusive tendency with Fe. Moreover, diffusion fluxes of V at the sediment-water interface illustrated that the release characteristics of V varied among the sampling sites. In addition, the BCR fraction of V in the surface sediments of the four sampling sites showed that V mainly existed in the residual fraction (88.04-88.57%). The concentrations of DGT-labile V (CDGT-V) were considerably lower than the non-residual fractions (the sum of exchangeable, reducible, and oxidizable fractions) measured by BCR sequential extraction. Correlation analysis showed that CDGT

  15. Maturation of the [Ni-4Fe-4S] active site of carbon monoxide dehydrogenases.

    PubMed

    Merrouch, Mériem; Benvenuti, Martino; Lorenzi, Marco; Léger, Christophe; Fourmond, Vincent; Dementin, Sébastien

    2018-02-14

    Nickel-containing enzymes are diverse in terms of function and active site structure. In many cases, the biosynthesis of the active site depends on accessory proteins which transport and insert the Ni ion. We review and discuss the literature related to the maturation of carbon monoxide dehydrogenases (CODH) which bear a nickel-containing active site consisting of a [Ni-4Fe-4S] center called the C-cluster. The maturation of this center has been much less studied than that of other nickel-containing enzymes such as urease and NiFe hydrogenase. Several proteins present in certain CODH operons, including the nickel-binding proteins CooT and CooJ, still have unclear functions. We question the conception that the maturation of all CODH depends on the accessory protein CooC described as essential for nickel insertion into the active site. The available literature reveals biological variations in CODH active site biosynthesis.

  16. First Partial Skeleton of a 1.34-Million-Year-Old Paranthropus boisei from Bed II, Olduvai Gorge, Tanzania

    PubMed Central

    Domínguez-Rodrigo, Manuel; Pickering, Travis Rayne; Baquedano, Enrique; Mabulla, Audax; Mark, Darren F.; Musiba, Charles; Bunn, Henry T.; Uribelarrea, David; Smith, Victoria; Diez-Martin, Fernando; Pérez-González, Alfredo; Sánchez, Policarpo; Santonja, Manuel; Barboni, Doris; Gidna, Agness; Ashley, Gail; Yravedra, José; Heaton, Jason L.; Arriaza, Maria Carmen

    2013-01-01

    Recent excavations in Level 4 at BK (Bed II, Olduvai Gorge, Tanzania) have yielded nine hominin teeth, a distal humerus fragment, a proximal radius with much of its shaft, a femur shaft, and a tibia shaft fragment (cataloged collectively as OH 80). Those elements identified more specifically than to simply Hominidae gen. et sp. indet are attributed to Paranthropus boisei. Before this study, incontrovertible P. boisei partial skeletons, for which postcranial remains occurred in association with taxonomically diagnostic craniodental remains, were unknown. Thus, OH 80 stands as the first unambiguous, dentally associated Paranthropus partial skeleton from East Africa. The morphology and size of its constituent parts suggest that the fossils derived from an extremely robust individual who, at 1.338±0.024 Ma (1 sigma), represents one of the most recent occurrences of Paranthropus before its extinction in East Africa. PMID:24339873

  17. CASPASE-9 CARD:CORE DOMAIN INTERACTIONS REQUIRE A PROPERLY-FORMED ACTIVE SITE

    PubMed Central

    Huber, Kristen L.; Serrano, Banyuhay P.; Hardy, Jeanne A.

    2018-01-01

    Caspase-9 is a critical factor in the initiation of apoptosis, and as a result is tightly regulated by a number of mechanisms. Caspase-9 contains a Caspase Activation and Recruitment Domain (CARD), which enables caspase-9 to form a tight interaction with the apoptosome, a heptameric activating platform. The caspase-9 CARD has been thought to be principally involved in recruitment to the apoptosome, but its roles outside this interaction have yet to be uncovered. In this work we show that the CARD is involved in physical interactions with the catalytic core of caspase-9 in the absence of the apoptosome; this interaction requires a properly formed caspase-9 active site. The active sites of caspases are composed of four extremely mobile loops. When the active-site loops are not properly ordered, the CARD and core domains of caspase-9 do not interact and behave independently, like loosely tethered beads. When the active-site loop bundle is properly ordered, the CARD domain interacts with the catalytic core, forming a single folding unit. Together these findings provide mechanistic insight into a new level of caspase-9 regulation, prompting speculation that the CARD may also play a role in the recruitment or recognition of substrate. PMID:29500231

  18. Prediction of active sites of enzymes by maximum relevance minimum redundancy (mRMR) feature selection.

    PubMed

    Gao, Yu-Fei; Li, Bi-Qing; Cai, Yu-Dong; Feng, Kai-Yan; Li, Zhan-Dong; Jiang, Yang

    2013-01-27

    Identification of catalytic residues plays a key role in understanding how enzymes work. Although numerous computational methods have been developed to predict catalytic residues and active sites, the prediction accuracy remains relatively low with high false positives. In this work, we developed a novel predictor based on the Random Forest algorithm (RF) aided by the maximum relevance minimum redundancy (mRMR) method and incremental feature selection (IFS). We incorporated features of physicochemical/biochemical properties, sequence conservation, residual disorder, secondary structure and solvent accessibility to predict active sites of enzymes and achieved an overall accuracy of 0.885687 and MCC of 0.689226 on an independent test dataset. Feature analysis showed that every category of the features except disorder contributed to the identification of active sites. It was also shown via the site-specific feature analysis that the features derived from the active site itself contributed most to the active site determination. Our prediction method may become a useful tool for identifying the active sites and the key features identified by the paper may provide valuable insights into the mechanism of catalysis.

  19. Active site densities, oxygen activation and adsorbed reactive oxygen in alcohol activation on npAu catalysts

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wang, Lu-Cun; Friend, C. M.; Fushimi, Rebecca

    The activation of molecular O 2as well as the reactivity of adsorbed oxygen species is of central importance in aerobic selective oxidation chemistry on Au-based catalysts. Herein, we address the issue of O 2activation on unsupported nanoporous gold (npAu) catalysts by applying a transient pressure technique, a temporal analysis of products (TAP) reactor, to measure the saturation coverage of atomic oxygen, its collisional dissociation probability, the activation barrier for O 2dissociation, and the facility with which adsorbed O species activate methanol, the initial step in the catalytic cycle of esterification. The results from these experiments indicate that molecular O 2dissociationmore » is associated with surface silver, that the density of reactive sites is quite low, that adsorbed oxygen atoms do not spill over from the sites of activation onto the surrounding surface, and that methanol reacts quite facilely with the adsorbed oxygen atoms. In addition, the O species from O 2dissociation exhibits reactivity for the selective oxidation of methanol but not for CO. The TAP experiments also revealed that the surface of the npAu catalyst is saturated with adsorbed O under steady state reaction conditions, at least for the pulse reaction.« less

  20. Active site densities, oxygen activation and adsorbed reactive oxygen in alcohol activation on npAu catalysts

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wang, Lu-Cun; Friend, C. M.; Fushimi, Rebecca

    2016-01-01

    The activation of molecular O 2as well as the reactivity of adsorbed oxygen species is of central importance in aerobic selective oxidation chemistry on Au-based catalysts. Herein, we address the issue of O 2activation on unsupported nanoporous gold (npAu) catalysts by applying a transient pressure technique, a temporal analysis of products (TAP) reactor, to measure the saturation coverage of atomic oxygen, its collisional dissociation probability, the activation barrier for O 2dissociation, and the facility with which adsorbed O species activate methanol, the initial step in the catalytic cycle of esterification. The results from these experiments indicate that molecular O 2dissociationmore » is associated with surface silver, that the density of reactive sites is quite low, that adsorbed oxygen atoms do not spill over from the sites of activation onto the surrounding surface, and that methanol reacts quite facilely with the adsorbed oxygen atoms. In addition, the O species from O 2dissociation exhibits reactivity for the selective oxidation of methanol but not for CO. The TAP experiments also revealed that the surface of the npAu catalyst is saturated with adsorbed O under steady state reaction conditions, at least for the pulse reaction.« less

  1. Brownian aggregation rate of colloid particles with several active sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nekrasov, Vyacheslav M.; Yurkin, Maxim A.; Chernyshev, Andrei V., E-mail: chern@ns.kinetics.nsc.ru

    2014-08-14

    We theoretically analyze the aggregation kinetics of colloid particles with several active sites. Such particles (so-called “patchy particles”) are well known as chemically anisotropic reactants, but the corresponding rate constant of their aggregation has not yet been established in a convenient analytical form. Using kinematic approximation for the diffusion problem, we derived an analytical formula for the diffusion-controlled reaction rate constant between two colloid particles (or clusters) with several small active sites under the following assumptions: the relative translational motion is Brownian diffusion, and the isotropic stochastic reorientation of each particle is Markovian and arbitrarily correlated. This formula was shownmore » to produce accurate results in comparison with more sophisticated approaches. Also, to account for the case of a low number of active sites per particle we used Monte Carlo stochastic algorithm based on Gillespie method. Simulations showed that such discrete model is required when this number is less than 10. Finally, we applied the developed approach to the simulation of immunoagglutination, assuming that the formed clusters have fractal structure.« less

  2. Stepwise Loop Insertion Strategy for Active Site Remodeling to Generate Novel Enzyme Functions.

    PubMed

    Hoque, Md Anarul; Zhang, Yong; Chen, Liuqing; Yang, Guangyu; Khatun, Mst Afroza; Chen, Haifeng; Hao, Liu; Feng, Yan

    2017-05-19

    The remodeling of active sites to generate novel biocatalysts is an attractive and challenging task. We developed a stepwise loop insertion strategy (StLois), in which randomized residue pairs are inserted into active site loops. The phosphotriesterase-like lactonase from Geobacillus kaustophilus (GkaP-PLL) was used to investigate StLois's potential for changing enzyme function. By inserting six residues into active site loop 7, the best variant ML7-B6 demonstrated a 16-fold further increase in catalytic efficiency toward ethyl-paraoxon compared with its initial template, that is a 609-fold higher, >10 7 fold substrate specificity shift relative to that of wild-type lactonase. The remodeled variants displayed 760-fold greater organophosphate hydrolysis activity toward the organophosphates parathion, diazinon, and chlorpyrifos. Structure and docking computations support the source of notably inverted enzyme specificity. Considering the fundamental importance of active site loops, the strategy has potential for the rapid generation of novel enzyme functions by loop remodeling.

  3. Orthogonal use of a human tRNA synthetase active site to achieve multifunctionality.

    PubMed

    Zhou, Quansheng; Kapoor, Mili; Guo, Min; Belani, Rajesh; Xu, Xiaoling; Kiosses, William B; Hanan, Melanie; Park, Chulho; Armour, Eva; Do, Minh-Ha; Nangle, Leslie A; Schimmel, Paul; Yang, Xiang-Lei

    2010-01-01

    Protein multifunctionality is an emerging explanation for the complexity of higher organisms. In this regard, aminoacyl tRNA synthetases catalyze amino acid activation for protein synthesis, but some also act in pathways for inflammation, angiogenesis and apoptosis. It is unclear how these multiple functions evolved and how they relate to the active site. Here structural modeling analysis, mutagenesis and cell-based functional studies show that the potent angiostatic, natural fragment of human tryptophanyl-tRNA synthetase (TrpRS) associates via tryptophan side chains that protrude from its cognate cellular receptor vascular endothelial cadherin (VE-cadherin). VE-cadherin's tryptophan side chains fit into the tryptophan-specific active site of the synthetase. Thus, specific side chains of the receptor mimic amino acid substrates and expand the functionality of the active site of the synthetase. We propose that orthogonal use of the same active site may be a general way to develop multifunctionality of human tRNA synthetases and other proteins.

  4. In silico analysis of Pycnoporus cinnabarinus laccase active site with toxic industrial dyes.

    PubMed

    Prasad, Nirmal K; Vindal, Vaibhav; Narayana, Siva Lakshmi; Ramakrishna, V; Kunal, Swaraj Priyaranjan; Srinivas, M

    2012-05-01

    Laccases belong to multicopper oxidases, a widespread class of enzymes implicated in many oxidative functions in various industrial oxidative processes like production of fine chemicals to bioremediation of contaminated soil and water. In order to understand the mechanisms of substrate binding and interaction between substrates and Pycnoporus cinnabarinus laccase, a homology model was generated. The resulted model was further validated and used for docking studies with toxic industrial dyes- acid blue 74, reactive black 5 and reactive blue 19. Interactions of chemical mediators with the laccase was also examined. The docking analysis showed that the active site always cannot accommodate the dye molecules, due to constricted nature of the active site pocket and steric hindrance of the residues whereas mediators are relatively small and can easily be accommodated into the active site pocket, which, thereafter leads to the productive binding. The binding properties of these compounds along with identification of critical active site residues can be used for further site-directed mutagenesis experiments in order to identify their role in activity and substrate specificity, ultimately leading to improved mutants for degradation of these toxic compounds.

  5. Earliest Porotic Hyperostosis on a 1.5-Million-Year-Old Hominin, Olduvai Gorge, Tanzania

    PubMed Central

    Domínguez-Rodrigo, Manuel; Pickering, Travis Rayne; Diez-Martín, Fernando; Mabulla, Audax; Musiba, Charles; Trancho, Gonzalo; Baquedano, Enrique; Bunn, Henry T.; Barboni, Doris; Santonja, Manuel; Uribelarrea, David; Ashley, Gail M.; Martínez-Ávila, María del Sol; Barba, Rebeca; Gidna, Agness; Yravedra, José; Arriaza, Carmen

    2012-01-01

    Meat-eating was an important factor affecting early hominin brain expansion, social organization and geographic movement. Stone tool butchery marks on ungulate fossils in several African archaeological assemblages demonstrate a significant level of carnivory by Pleistocene hominins, but the discovery at Olduvai Gorge of a child's pathological cranial fragments indicates that some hominins probably experienced scarcity of animal foods during various stages of their life histories. The child's parietal fragments, excavated from 1.5-million-year-old sediments, show porotic hyperostosis, a pathology associated with anemia. Nutritional deficiencies, including anemia, are most common at weaning, when children lose passive immunity received through their mothers' milk. Our results suggest, alternatively, that (1) the developmentally disruptive potential of weaning reached far beyond sedentary Holocene food-producing societies and into the early Pleistocene, or that (2) a hominin mother's meat-deficient diet negatively altered the nutritional content of her breast milk to the extent that her nursing child ultimately died from malnourishment. Either way, this discovery highlights that by at least 1.5 million years ago early human physiology was already adapted to a diet that included the regular consumption of meat. PMID:23056303

  6. Thermodynamic compensation upon binding to exosite 1 and the active site of thrombin

    PubMed Central

    Treuheit, Nicholas A.; Beach, Muneera A.; Komives, Elizabeth A.

    2011-01-01

    Several lines of experimental evidence including amide exchange and NMR suggest that ligands binding to thrombin cause reduced backbone dynamics. Binding of the covalent inhibitor dPhe-Pro-Arg chloromethylketone to the active site serine, as well as non-covalent binding of a fragment of the regulatory protein, thrombomodulin, to exosite 1 on the back side of the thrombin molecule both cause reduced dynamics. However, the reduced dynamics do not appear to be accompanied by significant conformational changes. In addition, binding of ligands to the active site does not change the affinity of thrombomodulin fragments binding to exosite 1, however, the thermodynamic coupling between exosite 1 and the active site has not been fully explored. We present isothermal titration calorimetry experiments that probe changes in enthalpy and entropy upon formation of binary ligand complexes. The approach relies on stringent thrombin preparation methods and on the use of dansyl-L-arginine-(3-methyl-1,5-pantanediyl) amide and a DNA aptamer as ligands with ideal thermodynamic signatures for binding to the active site and to exosite 1. Using this approach, the binding thermodynamic signatures of each ligand alone as well as the binding signatures of each ligand when the other binding site was occupied were measured. Different exosite 1 ligands with widely varied thermodynamic signatures cause the same reduction in ΔH and a concomitantly lower entropy cost upon DAPA binding at the active site. The results suggest a general phenomenon of enthalpy-entropy compensation consistent with reduction of dynamics/increased folding of thrombin upon ligand binding to either the active site or to exosite 1. PMID:21526769

  7. Thermodynamic compensation upon binding to exosite 1 and the active site of thrombin.

    PubMed

    Treuheit, Nicholas A; Beach, Muneera A; Komives, Elizabeth A

    2011-05-31

    Several lines of experimental evidence including amide exchange and NMR suggest that ligands binding to thrombin cause reduced backbone dynamics. Binding of the covalent inhibitor dPhe-Pro-Arg chloromethyl ketone to the active site serine, as well as noncovalent binding of a fragment of the regulatory protein, thrombomodulin, to exosite 1 on the back side of the thrombin molecule both cause reduced dynamics. However, the reduced dynamics do not appear to be accompanied by significant conformational changes. In addition, binding of ligands to the active site does not change the affinity of thrombomodulin fragments binding to exosite 1; however, the thermodynamic coupling between exosite 1 and the active site has not been fully explored. We present isothermal titration calorimetry experiments that probe changes in enthalpy and entropy upon formation of binary ligand complexes. The approach relies on stringent thrombin preparation methods and on the use of dansyl-l-arginine-(3-methyl-1,5-pantanediyl)amide and a DNA aptamer as ligands with ideal thermodynamic signatures for binding to the active site and to exosite 1. Using this approach, the binding thermodynamic signatures of each ligand alone as well as the binding signatures of each ligand when the other binding site was occupied were measured. Different exosite 1 ligands with widely varied thermodynamic signatures cause a similar reduction in ΔH and a concomitantly lower entropy cost upon DAPA binding at the active site. The results suggest a general phenomenon of enthalpy-entropy compensation consistent with reduction of dynamics/increased folding of thrombin upon ligand binding to either the active site or exosite 1.

  8. Active-Site Hydration and Water Diffusion in Cytochrome P450cam: A Highly Dynamic Process

    PubMed Central

    Miao, Yinglong; Baudry, Jerome

    2011-01-01

    Long-timescale molecular dynamics simulations (300 ns) are performed on both the apo- (i.e., camphor-free) and camphor-bound cytochrome P450cam (CYP101). Water diffusion into and out of the protein active site is observed without biased sampling methods. During the course of the molecular dynamics simulation, an average of 6.4 water molecules is observed in the camphor-binding site of the apo form, compared to zero water molecules in the binding site of the substrate-bound form, in agreement with the number of water molecules observed in crystal structures of the same species. However, as many as 12 water molecules can be present at a given time in the camphor-binding region of the active site in the case of apo-P450cam, revealing a highly dynamic process for hydration of the protein active site, with water molecules exchanging rapidly with the bulk solvent. Water molecules are also found to exchange locations frequently inside the active site, preferentially clustering in regions surrounding the water molecules observed in the crystal structure. Potential-of-mean-force calculations identify thermodynamically favored trans-protein pathways for the diffusion of water molecules between the protein active site and the bulk solvent. Binding of camphor in the active site modifies the free-energy landscape of P450cam channels toward favoring the diffusion of water molecules out of the protein active site. PMID:21943431

  9. Active Site Mutations Change the Cleavage Specificity of Neprilysin

    PubMed Central

    Sexton, Travis; Hitchcook, Lisa J.; Rodgers, David W.; Bradley, Luke H.; Hersh, Louis B.

    2012-01-01

    Neprilysin (NEP), a member of the M13 subgroup of the zinc-dependent endopeptidase family is a membrane bound peptidase capable of cleaving a variety of physiological peptides. We have generated a series of neprilysin variants containing mutations at either one of two active site residues, Phe563 and Ser546. Among the mutants studied in detail we observed changes in their activity towards leucine5-enkephalin, insulin B chain, and amyloid β1–40. For example, NEPF563I displayed an increase in preference towards cleaving leucine5-enkephalin relative to insulin B chain, while mutant NEPS546E was less discriminating than neprilysin. Mutants NEPF563L and NEPS546E exhibit different cleavage site preferences than neprilysin with insulin B chain and amyloid ß1–40 as substrates. These data indicate that it is possible to alter the cleavage site specificity of neprilysin opening the way for the development of substrate specific or substrate exclusive forms of the enzyme with enhanced therapeutic potential. PMID:22384224

  10. Carbinolamine Formation and Dehydration in a DNA Repair Enzyme Active Site

    PubMed Central

    Dodson, M. L.; Walker, Ross C.; Lloyd, R. Stephen

    2012-01-01

    In order to suggest detailed mechanistic hypotheses for the formation and dehydration of a key carbinolamine intermediate in the T4 pyrimidine dimer glycosylase (T4PDG) reaction, we have investigated these reactions using steered molecular dynamics with a coupled quantum mechanics–molecular mechanics potential (QM/MM). We carried out simulations of DNA abasic site carbinolamine formation with and without a water molecule restrained to remain within the active site quantum region. We recovered potentials of mean force (PMF) from thirty replicate reaction trajectories using Jarzynski averaging. We demonstrated feasible pathways involving water, as well as those independent of water participation. The water–independent enzyme–catalyzed reaction had a bias–corrected Jarzynski–average barrier height of approximately for the carbinolamine formation reaction and ) for the reverse reaction at this level of representation. When the proton transfer was facilitated with an intrinsic quantum water, the barrier height was approximately in the forward (formation) reaction and for the reverse. In addition, two modes of unsteered (free dynamics) carbinolamine dehydration were observed: in one, the quantum water participated as an intermediate proton transfer species, and in the other, the active site protonated glutamate hydrogen was directly transferred to the carbinolamine oxygen. Water–independent unforced proton transfer from the protonated active site glutamate carboxyl to the unprotonated N–terminal amine was also observed. In summary, complex proton transfer events, some involving water intermediates, were studied in QM/MM simulations of T4PDG bound to a DNA abasic site. Imine carbinolamine formation was characterized using steered QM/MM molecular dynamics. Dehydration of the carbinolamine intermediate to form the final imine product was observed in free, unsteered, QM/MM dynamics simulations, as was unforced acid-base transfer between the active site

  11. Immobilization of alkaline phosphatase on solid surface through self-assembled monolayer and by active-site protection.

    PubMed

    Gao, En-Feng; Kang, Kyung Lhi; Kim, Jeong Hee

    2014-06-01

    Retaining biological activity of a protein after immobilization is an important issue and many studies reported to enhance the activity of proteins after immobilization. We recently developed a new immobilization method of enzyme using active-site protection and minimization of the cross-links between enzyme and surface with a DNA polymerase as a model system. In this study, we extended the new method to an enzyme with a small mono-substrate using alkaline phosphatase (AP) as another model system. A condition to apply the new method is that masking agents, in this case its own substrate needs to stay at the active-site of the enzyme to be immobilized in order to protect the active-site during the harsh immobilization process. This could be achieved by removal of essential divalent ion, Zn2+ that is required for full enzyme activity of AP from the masking solution while active-site of AP was protected with p-nitrophenyl phosphate (pNPP). Approximately 40% of the solution-phase activity was acquired with active-site protected immobilized AP. In addition to protection active-site of AP, the number of immobilization links was kinetically controlled. When the mole fraction of the activated carboxyl group of the linker molecule in self-assembled monolayer (SAM) of 12-mercaptododecanoic acid and 6-mercapto-1-ethanol was varied, 10% of 12-mercaptododecanoic acid gave the maximum enzyme activity. Approximately 51% increase in enzyme activity of the active-site protected AP was observed compared to that of the unprotected group. It was shown that the concept of active-site protection and kinetic control of the number of covalent immobilization bonds can be extended to enzymes with small mono-substrates. It opens the possibility of further extension of the new methods of active-site protection and kinetic control of immobilization bond to important enzymes used in research and industrial fields.

  12. Ultrafast Light-Driven Substrate Expulsion from the Active Site of a Photoswitchable Catalyst.

    PubMed

    Pescher, Manuel D; van Wilderen, Luuk J G W; Grützner, Susanne; Slavov, Chavdar; Wachtveitl, Josef; Hecht, Stefan; Bredenbeck, Jens

    2017-09-25

    The photoswitchable piperidine general base catalyst is a prototype structure for light control of catalysis. Its azobenzene moiety moves sterically shielding groups to either protect or expose the active site, thereby changing the basicity and hydrogen-bonding affinity of the compound. The reversible switching dynamics of the catalyst is probed in the infrared spectral range by monitoring hydrogen bond (HB) formation between its active site and methanol (MeOH) as HB donor. Steady-state infrared (IR) and ultrafast IR and UV/Vis spectroscopies are used to uncover ultrafast expulsion of MeOH from the active site within a few picoseconds. Thus, the force generated by the azobenzene moiety even in the final phase of its isomerization is sufficient to break a strong HB within 3 ps and to shut down access to the active site. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Dynamic Perturbation of the Active Site Determines Reversible Thermal Inactivation in Glycoside Hydrolase Family 12.

    PubMed

    Jiang, Xukai; Li, Wen; Chen, Guanjun; Wang, Lushan

    2017-02-27

    The temperature dependence of enzyme catalysis is highly debated. Specifically, how high temperatures induce enzyme inactivation has broad implications for both fundamental and applied science. Here, we explored the mechanism of the reversible thermal inactivation in glycoside hydrolase family 12 (GH12) using comparative molecular dynamics simulations. First, we investigated the distribution of structural flexibility over the enzyme and found that the active site was the general thermal-sensitive region in GH12 cellulases. The dynamic perturbation of the active site before enzyme denaturation was explored through principal-component analysis, which indicated that variations in the collective motion and conformational ensemble of the active site may precisely correspond to enzyme transition from its active form to the inactive form. Furthermore, the degree of dynamic perturbation of the active site was found to be negatively correlated with the melting temperatures of GH12 enzymes, further proving the importance of the dynamic stability of the active site. Additionally, analysis of the residue-interaction network revealed that the active site in thermophilic enzyme was capable of forming additional contacts with other amino acids than those observed in the mesophilic enzyme. These interactions are likely the key mechanisms underlying the differences in rigidity of the active site. These findings provide further biophysical insights into the reversible thermal inactivation of enzymes and potential applications in future protein engineering.

  14. Non-active site mutation (Q123A) in New Delhi metallo-β-lactamase (NDM-1) enhanced its enzyme activity.

    PubMed

    Ali, Abid; Azam, Mohd W; Khan, Asad U

    2018-06-01

    New Delhi metallo β-lactamase-1 is one of the carbapenemases, causing hydrolysis of almost all β-lactamase antibiotics. Seventeen different NDM variants have been reported so far, they varied in their sequences either by single or multiple amino acid substitutions. Hence, it is important to understand its structural and functional relation. In the earlier studies role of active site residues has been studied but non-active site residues has not studied in detail. Therefore, we have initiated to further comprehend its structure and function relation by mutating some of its non-active site residues. A laboratory mutant of NDM-1 was generated by PCR-based site-directed mutagenesis, replacing Q to A at 123 position. The MICs of imipenem and meropenem for NDM-1 Q123A were found increased by 2 fold as compare to wild type and so the hydrolytic activity was enhanced (Kcat/Km) as compared to NDM-1 wild type. GOLD fitness scores were also found in favour of kinetics data. Secondary structure for α-helical content was determined by Far-UV circular dichroism (CD), which showed significant conformational changes. We conclude a noteworthy role of non-active-site amino acid residues in the catalytic activity of NDM-1. This study also provides an insight of emergence of new variants through natural evolution. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Multiple active site residues are important for photochemical efficiency in the light-activated enzyme protochlorophyllide oxidoreductase (POR).

    PubMed

    Menon, Binuraj R K; Hardman, Samantha J O; Scrutton, Nigel S; Heyes, Derren J

    2016-08-01

    Protochlorophyllide oxidoreductase (POR) catalyzes the light-driven reduction of protochlorophyllide (Pchlide), an essential, regulatory step in chlorophyll biosynthesis. The unique requirement of the enzyme for light has provided the opportunity to investigate how light energy can be harnessed to power biological catalysis and enzyme dynamics. Excited state interactions between the Pchlide molecule and the protein are known to drive the subsequent reaction chemistry. However, the structural features of POR and active site residues that are important for photochemistry and catalysis are currently unknown, because there is no crystal structure for POR. Here, we have used static and time-resolved spectroscopic measurements of a number of active site variants to study the role of a number of residues, which are located in the proposed NADPH/Pchlide binding site based on previous homology models, in the reaction mechanism of POR. Our findings, which are interpreted in the context of a new improved structural model, have identified several residues that are predicted to interact with the coenzyme or substrate. Several of the POR variants have a profound effect on the photochemistry, suggesting that multiple residues are important in stabilizing the excited state required for catalysis. Our work offers insight into how the POR active site geometry is finely tuned by multiple active site residues to support enzyme-mediated photochemistry and reduction of Pchlide, both of which are crucial to the existence of life on Earth. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Heterogeneous Electrocatalyst with Molecular Cobalt Ions Serving as the Center of Active Sites.

    PubMed

    Wang, Jiong; Ge, Xiaoming; Liu, Zhaolin; Thia, Larissa; Yan, Ya; Xiao, Wei; Wang, Xin

    2017-02-08

    Molecular Co 2+ ions were grafted onto doped graphene in a coordination environment, resulting in the formation of molecularly well-defined, highly active electrocatalytic sites at a heterogeneous interface for the oxygen evolution reaction (OER). The S dopants of graphene are suggested to be one of the binding sites and to be responsible for improving the intrinsic activity of the Co sites. The turnover frequency of such Co sites is greater than that of many Co-based nanostructures and IrO 2 catalysts. Through a series of carefully designed experiments, the pathway for the evolution of the Co cation-based molecular catalyst for the OER was further demonstrated on such a single Co-ion site for the first time. The Co 2+ ions were successively oxidized to Co 3+ and Co 4+ states prior to the OER. The sequential oxidation was coupled with the transfer of different numbers of protons/hydroxides and generated an active Co 4+ ═O fragment. A side-on hydroperoxo ligand of the Co 4+ site is proposed as a key intermediate for the formation of dioxygen.

  17. The response of mulberry trees after seedling hardening to summer drought in the hydro-fluctuation belt of Three Gorges Reservoir Areas.

    PubMed

    Huang, Xiaohui; Liu, Yun; Li, Jiaxing; Xiong, Xingzheng; Chen, Yang; Yin, Xiaohua; Feng, Dalan

    2013-10-01

    Interest has developed in the potential of mulberry (Morus alba), a woody perennial, for revegetating the hydro-fluctuation belt of the Three Gorges Reservoir due to its resistance to water-logging stress. To be useful, the trees must also be able to withstand dry conditions in summer when temperatures can be very high and droughts become severe quickly. Here, we report a study in which mulberry seedlings were grown in a greenhouse under a variety of simulated soil water conditions reflecting potential summer scenarios in the hydro-fluctuation belt of the Three Gorges Reservoir Area. We compared the responses of two pretreatment groups of mulberry seedlings to different levels of drought stress. The pretreatment groups differed with respect to drought hardening: the daily-managed (DM) group had relative soil moisture held constant in the range 70-80 %, while the drought-hardened (DH) group had relative soil moisture held constant at 40-50 %. Following the month-long pretreatment of seedlings, the two groups of young trees (DM and DH) were then respectively subjected to three levels of drought stress for a month: normal watering, moderate drought stress, and severe drought stress. A series of measurements comparing the physiological status of the plants in the two groups were then made, and the following results were obtained: (1) As drought stress increased, the heights, base diameters, root surface areas, photosynthetic rates (Pn), stomatal conductances (Gs), and transpiration rates (Tr) of the mulberry trees in both groups (DM and DH) decreased significantly, while the specific root area and abscisic acid (ABA) contents had increasing trends. Root activity and instantaneous water use efficiency of mulberry trees in both groups (DM and DH) were all raised under drought stress conditions than under normal watering, but the root/shoot ratio and leaf water potential were lowered. (2) At the same level of soil water content, the heights, base diameters, root

  18. Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor.

    PubMed

    Spatzal, Thomas; Perez, Kathryn A; Howard, James B; Rees, Douglas C

    2015-12-16

    Dinitrogen reduction in the biological nitrogen cycle is catalyzed by nitrogenase, a two-component metalloenzyme. Understanding of the transformation of the inert resting state of the active site FeMo-cofactor into an activated state capable of reducing dinitrogen remains elusive. Here we report the catalysis dependent, site-selective incorporation of selenium into the FeMo-cofactor from selenocyanate as a newly identified substrate and inhibitor. The 1.60 Å resolution structure reveals selenium occupying the S2B site of FeMo-cofactor in the Azotobacter vinelandii MoFe-protein, a position that was recently identified as the CO-binding site. The Se2B-labeled enzyme retains substrate reduction activity and marks the starting point for a crystallographic pulse-chase experiment of the active site during turnover. Through a series of crystal structures obtained at resolutions of 1.32-1.66 Å, including the CO-inhibited form of Av1-Se2B, the exchangeability of all three belt-sulfur sites is demonstrated, providing direct insights into unforeseen rearrangements of the metal center during catalysis.

  19. Composition and variability in the export of biogenic silica in the Changjiang River and the effect of Three Gorges Reservoir.

    PubMed

    Ran, Xiangbin; Liu, Sen; Liu, Jun; Zang, Jiaye; Che, Hong; Ma, Yongxing; Wang, Yibin

    2016-11-15

    Silicon (Si) plays an essential role in biogeochemical processes, but is still poorly characterized in the river system. This study addressed the biogenic silica (BSi) composition, origin and variation in the Changjiang River, and estimated the impacts of natural processes and human activities on the river Si cycling. Our results indicate that phytoliths comprised 14%-64% of BSi, while diatoms accounted for 34%-85% of BSi. The Changjiang River transported 620Ggyr(-1) of BSi and 2100Ggyr(-1) of dissolved silicate (DSi) loadings, respectively; 55% of the BSi and 51% of the DSi fluxes are transported during the high discharge period from June to September. The Changjiang River carried phytolith BSi mostly comes from the middle and lower reaches area. The ratio of BSi/(BSi+DSi) has decreased from 0.47 before 1980 to 0.19 in 2013-2014 due to the direct retention of BSi. The BSi sedimentation in the Three Gorges Reservoir would cause a decrease of total reactive silica, but contribute to approximately 4%-16% of the DSi loading at the Jiangyin station due to its dissolution. This study demonstrates that phytoliths represent a significant contribution to the biogeochemical cycle of silica in coastal waters, and in-stream process exerts a great influence on the river Si loading and cycling. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. The impact of lake level variation on seismicity around XianNvShan fault in the Three Gorge area

    NASA Astrophysics Data System (ADS)

    Liao, W.; Li, J.; Zhang, L.

    2017-12-01

    Since the impounding of Three Gorge Project in 2003,more than 10000 earthquakes have been recorded by the digital telemetry seismic network. Most of them occurred around the GaoQiao fault and the Northern segment of XianNvShan fault . In March 2014, the M4.3 and M4.7 earthquake happened in the northern segment of Xiannvshshan fault .In order to study the relationship between the seismicity around the XianNvShan fault and the lake level variation, we had been deployed 5 temporal seismic stations in this area from 2015 to 2016. More than 3000 earthquakes recorded during the time of temporal seismic monitoring are located by hypo-center of by waveform cross-correlation and double-difference method. The depth of most earthquakes is from 5 to 7 km.but it is obvious that the variation of depth is relate to the fluctuation of water level.

  1. Chemical quality and temperature of water in Flaming Gorge Reservoir, Wyoming and Utah, and the effect of the reservoir on the Green River

    USGS Publications Warehouse

    Bolke, E.L.; Waddell, Kidd M.

    1975-01-01

    The major tributaries to Flaming Gorge Reservoir contribute an average of about 97 percent of the total streamflow and 82 percent of the total load of dissolved solids. The Green River is the largest tributary, and for the 1957-72 water years it contributed 81 percent of the total streamflow and 70 percent of the total load of dissolved solids. The principal constituents in the tributary streamflow are calcium and sulfate during periods of lowest flow and calcium and bicarbonate during periods of highest flow.Flaming Gorge Dam was closed in November 1962, and the most significant load changes of chemical constituents due to the net effect of inflow, outflow, leaching, and chemical precipitation in the reservoir have been load changes of sulfate and bicarbonate. The average increase of dissolved load of sulfate in the reservoir for the 1969-72 water years was 110,000 tons (99,790 t) per year, which was 40,000 tons (36,287 t) per year less than for the 1963-66 water years. The average decrease of dissolved load of bicarbonate in the reservoir for 1969-72 was 40,000 tons (36,287 t) per year, which was the same as the decrease for 1963-66.Anaerobic conditions were observed in the deep, uncirculated part of the reservoir near the dam during the 1971 and 1972 water years, and anaerobic or near-anaerobic conditions were observed near the confluence of the Blacks Fork and Green River during the summers of 1971 and 1972.The water in Flaming Gorge Reservoir is in three distinct layers, and the upper two layers (the epilimnion and the metalimnion) mixed twice during each of the 1971-72 water years. The two circulation periods were in the spring and fall. The water in the deepest layer (the hypolimnion) did not mix with the waters of the upper zones because the density difference was too great and because the deep, narrow shape of the basin probably inhibits mixing.The depletion of flow in the Green River downstream from Flaming Gorge Dam between closure of the dam and the end

  2. Methylation of an alpha-foetoprotein gene intragenic site modulates gene activity.

    PubMed Central

    Opdecamp, K; Rivière, M; Molné, M; Szpirer, J; Szpirer, C

    1992-01-01

    By comparing the methylation pattern of Mspl/Hpall sites in the 5' region of the mouse alpha-foetoprotein (AFP) gene of different cells (hepatoma cells, foetal and adult liver, fibroblasts), we found a correlation between gene expression and unmethylation of a site located in the first intron of the gene. Other sites did not show this correlation. In transfection experiments of unmethylated and methylated AFP-CAT chimeric constructions, we then showed that methylation of the intronic site negatively modulates expression of CAT activity. We also found that a DNA segment centered on this site binds nuclear proteins; however methylation did not affect protein binding. Images PMID:1371343

  3. Molecular imprint of enzyme active site by camel nanobodies: rapid and efficient approach to produce abzymes with alliinase activity.

    PubMed

    Li, Jiang-Wei; Xia, Lijie; Su, Youhong; Liu, Hongchun; Xia, Xueqing; Lu, Qinxia; Yang, Chunjin; Reheman, Kalbinur

    2012-04-20

    Screening of inhibitory Ab1 antibodies is a critical step for producing catalytic antibodies in the anti-idiotypic approach. However, the incompatible surface of the active site of the enzyme and the antigen-binding site of heterotetrameric conventional antibodies become the limiting step. Because camelid-derived nanobodies possess the potential to preferentially bind to the active site of enzymes due to their small size and long CDR3, we have developed a novel approach to produce antibodies with alliinase activities by exploiting the molecular mimicry of camel nanobodies. By screening the camelid-derived variable region of the heavy chain cDNA phage display library with alliinase, we obtained an inhibitory nanobody VHHA4 that recognizes the active site. Further screening with VHHA4 from the same variable domain of the heavy chain of a heavy-chain antibody library led to a higher incidence of anti-idiotypic Ab2 abzymes with alliinase activities. One of the abzymes, VHHC10, showed the highest activity that can be inhibited by Ab1 VHHA4 and alliinase competitive inhibitor penicillamine and significantly suppressed the B16 tumor cell growth in the presence of alliin in vitro. The results highlight the feasibility of producing abzymes via anti-idiotypic nanobody approach.

  4. Soil stability characteristics of mulberry lands at hydro-fluctuation belt in the Three Gorges Reservoir area, China.

    PubMed

    Jiang, Ping; Shi, Dongmei; Hu, Xueqin; Huang, Xianzhi; Li, Yexin; Guo, Tianlei

    2015-10-01

    The hydro-fluctuation belt in the Three Gorges Reservoir area is a typical seasonal and artificial wetland system and ecologically fragile zone. Using the widely existing mulberry forest lands in the hydro-fluctuation belt as an example and the 180-m water-level forest land as a control, this paper analyzes the soil stability of mulberry forestlands at different water levels in the hydro-fluctuation belt by analyzing and comparing the changes between soil physical and mechanical properties. The results indicated that (1) water-level changes, such as rising, flooding, draining, and exposure, affect the soil structure in mulberry forestlands. The soil agglomeration statuses for the soil layers decreased from 180 > 175 > 170 > 165 m, and the soil agglomeration statuses at a depth of 0∼20 cm decreased by 43.79, 44.95, and 57.45% compared with the control. (2) The soil water stability index decreased as follows: 180 > 170 > 175 > 165 m, which only accounted for 50.00, 47.73, and 40.91% of the control. In addition, the soil water stability indexes for the topsoils at various water levels were 1.87 (180 m), 1.67 (175 m), 2.92 (170 m), and 1.86 (165 m) times greater than those of the subsoils; thus, the resistance to hydraulic dispersion and disintegration were greater in the topsoil than in the subsoil. (3) The soil aggregate stability index decreased from 180 > 165 > 170 > 175 m and by 22.75, 23.53, and 35.29% compared with the control. (4) The soil shear strengths (composed of the cohesive force C and the internal friction angle φ) of the topsoils at water levels of 175, 170, and 165 m were significantly lower than in the control, and the internal friction angles decreased by 10.52, 19.08, and 43.25% and the cohesive force decreased by 9.88, 16.36, and 27.51%, respectively. The stability of the soil structure was greatly influenced by the soil clay content, soil organic matter content, and waterlogging duration. The study results

  5. Synthesis of a molecularly defined single-active site heterogeneous catalyst for selective oxidation of N-heterocycles.

    PubMed

    Zhang, Yujing; Pang, Shaofeng; Wei, Zhihong; Jiao, Haijun; Dai, Xingchao; Wang, Hongli; Shi, Feng

    2018-04-13

    Generally, a homogeneous catalyst exhibits good activity and defined active sites but it is difficult to recycle. Meanwhile, a heterogeneous catalyst can easily be reused but its active site is difficult to reveal. It is interesting to bridge the gap between homogeneous and heterogeneous catalysis via controllable construction of a heterogeneous catalyst containing defined active sites. Here, we report that a molecularly defined, single-active site heterogeneous catalyst has been designed and prepared via the oxidative polymerization of maleimide derivatives. These polymaleimide derivatives can be active catalysts for the selective oxidation of heterocyclic compounds to quinoline and indole via the recycling of -C=O and -C-OH groups, which was confirmed by tracing the reaction with GC-MS using maleimide as the catalyst and by FT-IR analysis with polymaleimide as the catalyst. These results might promote the development of heterogeneous catalysts with molecularly defined single active sites exhibiting a comparable activity to homogeneous catalysts.

  6. Effects of the human activities on the water level process of the Poyang Lake

    NASA Astrophysics Data System (ADS)

    Zhao, Jun-kai; Chen, Li; Yang, Yun-xian

    2017-12-01

    The hydrological cycles in basin is profoundly affected by human activities. Yangtze River is a world class river with complex river-lake relations in the middle reaches. As the Three Gorges Reservoir (TGR) and other controlled reservoirs in the main stream and tributaries have been put into operation, the water regimes of the main stream in the middle reaches and Poyang Lake have been changed by water impounding and sediments trapping, clean water discharged from reservoirs, accelerating the evolution of the relationship of river and lake. After entering the 21st century, autumn droughts become more serious in Poyang Lake basin; the relationship between river and lake becomes tense. In light of the hydrological data in Poyang Lake since 2000s, this article made quantitative analyses of the influences of the human activities on the variation of the Poyang Lake level by authors. The results indicate that the main stream of Yangtze River, particularly the regulation of Three Gorges Reservoir, exerts a profound influence on the variation process of the Poyang Lake level. The regulation influence of the Upper Reach of the Yangtze River’s Reservoir Group (URYRRG) could spread to Tangyin area in the middle of the lake in October.

  7. Biological role of site-specific O-glycosylation in cell adhesion activity and phosphorylation of osteopontin.

    PubMed

    Oyama, Midori; Kariya, Yoshinobu; Kariya, Yukiko; Matsumoto, Kana; Kanno, Mayumi; Yamaguchi, Yoshiki; Hashimoto, Yasuhiro

    2018-05-09

    Osteopontin (OPN) is an extracellular glycosylated phosphoprotein that promotes cell adhesion by interacting with several integrin receptors. We previously reported that an OPN mutant lacking five O-glycosylation sites (Thr 134 /Thr 138 /Thr 143 /Thr 147 /Thr 152 ) in the threonine/proline-rich region increased cell adhesion activity and phosphorylation compared with the wild type. However, the role of O-glycosylation in cell adhesion activity and phosphorylation of OPN remains to be clarified. Here, we show that site-specific O-glycosylation in the threonine/proline-rich region of OPN affects its cell adhesion activity and phosphorylation independently and/or synergistically. Using site-directed mutagenesis, we found that OPN mutants with substitution sets of Thr 134 /Thr 138 or Thr 143 /Thr 147 /Thr 152 had decreased and increased cell adhesion activity, respectively. In contrast, the introduction of a single mutation into the O-glycosylation sites had no effect on OPN cell adhesion activity. An adhesion assay using function-blocking antibodies against αvβ3 and β1 integrins, as well as αvβ3 integrin-overexpressing A549 cells, revealed that site-specific O-glycosylation affected the association of OPN with the two integrins. Phosphorylation analyses using phos-tag and LC-MS/MS indicated that phosphorylation levels and sites were influenced by the O-glycosylation status, although the number of O-glycosylation sites was not correlated with the phosphorylation level in OPN. Furthermore, a correlation analysis between phosphorylation level and cell adhesion activity in OPN mutants with the site-specific O-glycosylation showed that they were not always correlated. These results provide conclusive evidence of a novel regulatory mechanism of cell adhesion activity and phosphorylation of OPN by site-specific O-glycosylation. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  8. Interactive flare sites within an active region complex

    NASA Technical Reports Server (NTRS)

    Poletto, G.; Gary, G. A.; Machado, M. E.

    1993-01-01

    We examine here a set of images of an active region complex, acquired on June 24-25, 1980, by the Hard X-ray Imaging Spectrometer on SMM, with the purpose of establishing whether there was any interplay between the frequent activity observed at different sites in the activity center and, in such a case, how the interaction was established. By analyzing both quiet and active orbits we show that, as a rule, activity originating in one region triggers the other region's activity. However, we find little unambiguous evidence for the presence of large-scale interconnecting loops. A comparison of X-ray images with magnetic field observations suggested that we interpret the active region behavior in terms of the interaction between different loop systems, in a scenario quite analogous to the interacting bipole representation of individual flares. We conclude that active region interplay provides an easily observable case to study the time-dependent topology and the mechanisms for the spreading of activity in transient events over all energy scales.

  9. Ultrafast infrared spectroscopy reveals water-mediated coherent dynamics in an enzyme active site.

    PubMed

    Adamczyk, Katrin; Simpson, Niall; Greetham, Gregory M; Gumiero, Andrea; Walsh, Martin A; Towrie, Michael; Parker, Anthony W; Hunt, Neil T

    2015-01-01

    Understanding the impact of fast dynamics upon the chemical processes occurring within the active sites of proteins and enzymes is a key challenge that continues to attract significant interest, though direct experimental insight in the solution phase remains sparse. Similar gaps in our knowledge exist in understanding the role played by water, either as a solvent or as a structural/dynamic component of the active site. In order to investigate further the potential biological roles of water, we have employed ultrafast multidimensional infrared spectroscopy experiments that directly probe the structural and vibrational dynamics of NO bound to the ferric haem of the catalase enzyme from Corynebacterium glutamicum in both H 2 O and D 2 O. Despite catalases having what is believed to be a solvent-inaccessible active site, an isotopic dependence of the spectral diffusion and vibrational lifetime parameters of the NO stretching vibration are observed, indicating that water molecules interact directly with the haem ligand. Furthermore, IR pump-probe data feature oscillations originating from the preparation of a coherent superposition of low-frequency vibrational modes in the active site of catalase that are coupled to the haem ligand stretching vibration. Comparisons with an exemplar of the closely-related peroxidase enzyme family shows that they too exhibit solvent-dependent active-site dynamics, supporting the presence of interactions between the haem ligand and water molecules in the active sites of both catalases and peroxidases that may be linked to proton transfer events leading to the formation of the ferryl intermediate Compound I. In addition, a strong, water-mediated, hydrogen bonding structure is suggested to occur in catalase that is not replicated in peroxidase; an observation that may shed light on the origins of the different functions of the two enzymes.

  10. [Concentrations and pollution assessment of soil heavy metals at different water-level altitudes in the draw-down areas of the Three Gorges Reservoir].

    PubMed

    Wang, Ye-Chun; Lei, Bo; Yang, San-Ming; Zhang, Sheng

    2012-02-01

    To investigate the effect of 175 m trial impounding (2008 and 2009) of the Three Gorges Reservoir on soil heavy metals, three draw-down areas with similar geological environment and history of land-use in Zhongxian County were chosen. Altogether 36 surface soil samples (including 0-10 cm and 10-20 cm soil layer) from water-level altitude of 160 m and 170 m were obtained, and their heavy metals concentrations (As, Cd, Cr, Cu, Ni, Pb and Zn) were measured by the X-ray fluorescence spectrometric method. Geoaccumulation index (I(geo)) and Håkanson potential ecological risk index were applied to assess the heavy metals pollution status and potential ecological risk, respectively. Results indicated that although the inundation period of 160 m was 224 d longer than that of 170 m, significant difference in concentrations of heavy metals were not found between the two water-level altitudes. Except for Cd, most of the heavy metals highly related with each other positively. According to the geoaccumulation index, the pollution extent of the heavy metals followed the order: As > Cd > Cu > Ni > Zn = Pb > Cr. The I(geo) value of As, Cd and Cu were 0.45, 0.39 and 0.06, respectively, indicating that the soil was only lightly polluted by these heavy metals. Håkanson single potential ecological risk index followed the order: Cd > As > Cu > Pb > Ni > Cr > Zn. Cd with E(i) values of 59.10, had a medium potential for ecological risk,while As, Cr, Cu, Pb, Ni and Zn only had a light potential. Consequently, although As, Cd and Cu were the major heavy metals with potential ecological risk for surface soil pollution in the draw-down areas in Zhongxian County, the Three Gorges Reservoir.

  11. Occurrence and Characteristics of Microplastic Pollution in Xiangxi Bay of Three Gorges Reservoir, China.

    PubMed

    Zhang, Kai; Xiong, Xiong; Hu, Hongjuan; Wu, Chenxi; Bi, Yonghong; Wu, Yonghong; Zhou, Bingsheng; Lam, Paul K S; Liu, Jiantong

    2017-04-04

    Microplastic pollution in inland waters is receiving growing attentions. Reservoirs are suspected to be particularly vulnerable to microplastic pollution. However, very limited information is currently available on pollution characteristics of microplastics in reservoir ecosystems. This work studied the distribution and characteristics of microplastics in the backwater area of Xiangxi River, a typical tributary of the Three Gorges Reservoir. Microplastics were detected in both surface water and sediment with concentrations ranging from 0.55 × 10 5 to 342 × 10 5 items km -2 and 80 to 864 items m -2 , respectively. Polyethylene, polypropylene, and polystyrene were identified in surface water, whereas polyethylene, polypropylene, and polyethylene terephthalate, and pigments were observed in sediment. In addition, microplastics were also detected in the digestion tracts of 25.7% of fish samples, and polyethylene and nylon were identified. Redundancy analysis indicates a weak correlation between microplastics and water quality variables but a negative correlation with water level of the reservoir and Secchi depth. Results from this study confirm the presence of high abundance microplastics in reservoir impacted tributaries, and suggest that water level regulated hydrodynamic condition and input of nonpoint sources are important regulators for microplastic accumulation and distribution in the backwater area of reservoir tributaries.

  12. Engineering streptokinase for generation of active site-labeled plasminogen analogs*

    PubMed Central

    Laha, Malabika; Panizzi, Peter; Nahrendorf, Matthias; Bock, Paul E.

    2011-01-01

    We previously demonstrated that streptokinase (SK) can be used to generate active site-labeled fluorescent analogs of plasminogen (Pg) by virtue of its non-proteolytic activation of the zymogen. The method is versatile and allows for stoichiometric and active site-specific incorporation of any one of many molecular probes. The limitation of the labeling approach is that it is both time-consuming and low yield. Here we demonstrate an improved method for the preparation of labeled Pg analogs by the use of an engineered SK mutant fusion protein with both COOH- and NH2-terminal His6-tags. The NH2-terminal tag is followed by a tobacco etch virus proteinase cleavage site to ensure that the SK Ile1 residue, essential for conformational activation of Pg, is preserved. The SK COOH-terminal Lys414 residue and residues Arg253-Leu260 in the SK β-domain were deleted to prevent cleavage by plasmin (Pm), and to disable Pg substrate binding to the SK·Pg*/Pm catalytic complexes, respectively. Near-elimination of Pm generation with the SKΔ(R253-L260)ΔK414-His6 mutant increased the yield of labeled Pg 2.6-fold and reduced the time required >2-fold. The versatility of the labeling method was extended to the application of Pg labeled with a near-infrared probe to quantitate Pg receptors on immune cells by flow cytometry. PMID:21570944

  13. Quantifying the density and utilization of active sites in non-precious metal oxygen electroreduction catalysts

    PubMed Central

    Sahraie, Nastaran Ranjbar; Kramm, Ulrike I.; Steinberg, Julian; Zhang, Yuanjian; Thomas, Arne; Reier, Tobias; Paraknowitsch, Jens-Peter; Strasser, Peter

    2015-01-01

    Carbon materials doped with transition metal and nitrogen are highly active, non-precious metal catalysts for the electrochemical conversion of molecular oxygen in fuel cells, metal air batteries, and electrolytic processes. However, accurate measurement of their intrinsic turn-over frequency and active-site density based on metal centres in bulk and surface has remained difficult to date, which has hampered a more rational catalyst design. Here we report a successful quantification of bulk and surface-based active-site density and associated turn-over frequency values of mono- and bimetallic Fe/N-doped carbons using a combination of chemisorption, desorption and 57Fe Mössbauer spectroscopy techniques. Our general approach yields an experimental descriptor for the intrinsic activity and the active-site utilization, aiding in the catalyst development process and enabling a previously unachieved level of understanding of reactivity trends owing to a deconvolution of site density and intrinsic activity. PMID:26486465

  14. Exploring the Active Site of the Tungsten, Iron-Sulfur Enzyme Acetylene Hydratase▿ †

    PubMed Central

    tenBrink, Felix; Schink, Bernhard; Kroneck, Peter M. H.

    2011-01-01

    The soluble tungsten, iron-sulfur enzyme acetylene hydratase (AH) from mesophilic Pelobacter acetylenicus is a member of the dimethyl sulfoxide (DMSO) reductase family. It stands out from its class as it catalyzes a nonredox reaction, the addition of H2O to acetylene (H—C☰C—H) to form acetaldehyde (CH3CHO). Caught in its active W(IV) state, the high-resolution three-dimensional structure of AH offers an excellent starting point to tackle its unique chemistry and to identify catalytic amino acid residues within the active site cavity: Asp13 close to W(IV) coordinated to two molybdopterin-guanosine-dinucleotide ligands, Lys48 which couples the [4Fe-4S] cluster to the W site, and Ile142 as part of a hydrophobic ring at the end of the substrate access channel designed to accommodate the substrate acetylene. A protocol was developed to express AH in Escherichia coli and to produce active-site variants which were characterized with regard to activity and occupancy of the tungsten and iron-sulfur centers. By this means, fusion of the N-terminal chaperone binding site of the E. coli nitrate reductase NarG to the AH gene improved the yield and activity of AH and its variants significantly. Results from site-directed mutagenesis of three key residues, Asp13, Lys48, and Ile142, document their important role in catalysis of this unusual tungsten enzyme. PMID:21193613

  15. Comparison of the active-site design of molybdenum oxo-transfer enzymes by quantum mechanical calculations.

    PubMed

    Li, Jilai; Ryde, Ulf

    2014-11-17

    There are three families of mononuclear molybdenum enzymes that catalyze oxygen atom transfer (OAT) reactions, named after a typical example from each family, viz., dimethyl sulfoxide reductase (DMSOR), sulfite oxidase (SO), and xanthine oxidase (XO). These families differ in the construction of their active sites, with two molybdopterin groups in the DMSOR family, two oxy groups in the SO family, and a sulfido group in the XO family. We have employed density functional theory calculations on cluster models of the active sites to understand the selection of molybdenum ligands in the three enzyme families. Our calculations show that the DMSOR active site has a much stronger oxidative power than the other two sites, owing to the extra molybdopterin ligand. However, the active sites do not seem to have been constructed to make the OAT reaction as exergonic as possible, but instead to keep the reaction free energy close to zero (to avoid excessive loss of energy), thereby making the reoxidation (SO and XO) or rereduction of the active sites (DMSOR) after the OAT reaction facile. We also show that active-site models of the three enzyme families can all catalyze the reduction of DMSO and that the DMSOR model does not give the lowest activation barrier. Likewise, all three models can catalyze the oxidation of sulfite, provided that the Coulombic repulsion between the substrate and the enzyme model can be overcome, but for this harder reaction, the SO model gives the lowest activation barrier, although the differences are not large. However, only the XO model can catalyze the oxidation of xanthine, owing to its sulfido ligand.

  16. Enzyme/non-enzyme discrimination and prediction of enzyme active site location using charge-based methods.

    PubMed

    Bate, Paul; Warwicker, Jim

    2004-07-02

    Calculations of charge interactions complement analysis of a characterised active site, rationalising pH-dependence of activity and transition state stabilisation. Prediction of active site location through large DeltapK(a)s or electrostatic strain is relevant for structural genomics. We report a study of ionisable groups in a set of 20 enzymes, finding that false positives obscure predictive potential. In a larger set of 156 enzymes, peaks in solvent-space electrostatic properties are calculated. Both electric field and potential match well to active site location. The best correlation is found with electrostatic potential calculated from uniform charge density over enzyme volume, rather than from assignment of a standard atom-specific charge set. Studying a shell around each molecule, for 77% of enzymes the potential peak is within that 5% of the shell closest to the active site centre, and 86% within 10%. Active site identification by largest cleft, also with projection onto a shell, gives 58% of enzymes for which the centre of the largest cleft lies within 5% of the active site, and 70% within 10%. Dielectric boundary conditions emphasise clefts in the uniform charge density method, which is suited to recognition of binding pockets embedded within larger clefts. The variation of peak potential with distance from active site, and comparison between enzyme and non-enzyme sets, gives an optimal threshold distinguishing enzyme from non-enzyme. We find that 87% of the enzyme set exceeds the threshold as compared to 29% of the non-enzyme set. Enzyme/non-enzyme homologues, "structural genomics" annotated proteins and catalytic/non-catalytic RNAs are studied in this context.

  17. Soil moisture status estimation over Three Gorges area with Landsat TM data based on temperature vegetation dryness index

    NASA Astrophysics Data System (ADS)

    Xu, Lina; Niu, Ruiqing; Li, Jiong; Dong, Yanfang

    2011-12-01

    Soil moisture is the important indicator of climate, hydrology, ecology, agriculture and other parameters of the land surface and atmospheric interface. Soil moisture plays an important role on the water and energy exchange at the land surface/atmosphere interface. Remote sensing can provide information on large area quickly and easily, so it is significant to do research on how to monitor soil moisture by remote sensing. This paper presents a method to assess soil moisture status using Landsat TM data over Three Gorges area in China based on TVDI. The potential of Temperature- Vegetation Dryness Index (TVDI) from Landsat TM data in assessing soil moisture was investigated in this region. After retrieving land surface temperature and vegetation index a TVDI model based on the features of Ts-NDVI space is established. And finally, soil moisture status is estimated according to TVDI. It shows that TVDI has the advantages of stability and high accuracy to estimating the soil moisture status.

  18. The Sasso Pizzuto landslide dam and seismically induced rockfalls along the Nera River gorge (Central Italy).

    NASA Astrophysics Data System (ADS)

    Romeo, Saverio; Di Matteo, Lucio; Melelli, Laura; Cencetti, Corrado; Dragoni, Walter; Fredduzzi, Andrea; De Rosa, Pierluigi

    2017-04-01

    The seismically induced landslides are among the most destructive and dangerous effects of an earthquake. In the Italian contest, this is also documented by a national catalogue that collects data related to earthquake-induced ground failures in the last millennium (CEDIT database). In particular, Central Italy has been affected by several historical landslides triggered by significant earthquakes, the last of which occurred in August-October 2016, representing the Italian strongest event after the 1980 Irpinia earthquake (Mw 6.9). The study presents the effects of recent seismically induced rockfalls occurred within the Central Italy seismic sequence (October 30, 2016) along the Nera River gorge between Umbria and Marche. The study area is completely included in the Monti Sibillini National Park, where the highest mountain chain in the Umbrian-Marchean Apennine is located. Most of rockfalls have affected the "Maiolica" formation, a stratified and fractured pelagic limestone dating to the Early Cretaceous. The seismic sequence produced diffuse instabilities along the SP 209 road within the Nera River gorge: boulders, debris accumulations and diffuse rockfalls have been mapped. Most of boulders have size ranging from 0.3 to 2.0 m in diameter. Although several strong quakes (Mw > 5) occurred during the August-October sequence, only the main quake triggered the Sasso Pizzuto rockfall producing a landslide dam along the Nera River. The landslide appears to have originated as a wedge failure, which evolved to free fall when the rock block lost the contact with the stable rock mass. In other words, the quake produced the "explosion" of the rock wall allowing the rockfall process. Once the rock mass reached the toe of the slope, it was broken triggering a rock avalanche that obstructed both the Nera River and SP 209 road. With the aim to estimate the total volume of involved rock, a field survey was carried out by using a laser rangefinder. Remote measures were acquired

  19. Plant soaking decomposition as well as nitrogen and phosphorous release in the water-level fluctuation zone of the Three Gorges Reservoir.

    PubMed

    Xiao, Liwei; Zhu, Bo; Nsenga Kumwimba, Mathieu; Jiang, Shiwei

    2017-08-15

    The operating scheme of the Three Gorges Reservoir results in a summer drought in the water-level fluctuation zone during which plants grow vigorously. In the winter inundation season, soaking plants may decompose and release nutrients resulting in water quality deterioration. This study quantifies the contributions of the underwater decomposition of nine dominant plant species in the water-level fluctuation zone to nutrient release. The in-situ litterbag technique was used to study for soaking decomposition over 200days. All soaking plant species decomposed rapidly at an average rate of 1.99±0.33%d -1 in the early stage of soaking (0 to 30days) and at an average rate of only 0.07±0.04%d -1 in the later stage (30 to 200days). After 200days of soaking, the nine plant species released an average of 312.40±39.97gkg -1 organic carbon, 6.71±4.29gkg -1 of nitrogen and 2.25±1.25gkg -1 of phosphorus. A positive relationship was found between soaking plant decomposition rates and initial C/N ratios of 25 to 50, and a negative relationship where the C/N ratios were between 50 and 100. The amounts of total nitrogen or total phosphorus released were significantly negatively correlated with the initial C/N or C/P ratios of the plants. Among the studied plant species, Xanthium sibiricum Patr ex Widder showed high level of nutrient release via soaking decomposition. In contrast, Cynodon dactylon (Linn.) Pers. and Polygonum hydropiper exhibited low levels of nutrient release and are recommended as suitable species for the ecological restoration of the water-level fluctuation zone. Our results demonstrate that after 200days of soaking plant decomposition, the loadings of total organic carbon, nitrogen, and phosphorus in the water-level fluctuation zone of the Three Gorges Reservoir were 2942.1, 81.1, and 24.7kgha -1 , respectively and therefore could potentially damage the aquatic environment of the reservoir. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Active Site Flexibility of Mycobacterium tuberculosis Isocitrate Lyase in Dimer Form.

    PubMed

    Lee, Yie-Vern; Choi, Sy Bing; Wahab, Habibah A; Choong, Yee Siew

    2017-09-25

    Tuberculosis (TB) still remains a global threat due to the emergence of a drug-resistant strain. Instead of focusing on the drug target of active stage TB, we are highlighting the isocitrate lyase (ICL) at the dormant stage TB. ICL is one of the persistent factors for Mycobacterium tuberculosis (MTB) to survive during the dormant phase. In addition, the absence of ICL in human has made ICL a potential drug target for TB therapy. However, the dynamic details of ICL which could give insights to the ICL-ligand interaction have yet to be solved. Therefore, a series of ICL dimer dynamics studies through molecular dynamics simulation were performed in this work. The ICL active site entrance gate closure is contributed to by hydrogen bonding and electrostatic interactions with the C-terminal. Analysis suggested that the open-closed behavior of the ICL active site entrance depends on the type of ligand present in the active site. We also observed four residues (Ser91, Asp108, Asp153, and Cys191) which could possibly be the nucleophiles for nucleophilic attack on the cleavage of isocitrate at the C 2 -C 3 bond. We hope that the elucidation of ICL dynamics can benefit future works such as lead identification or antibody design against ICL for TB therapeutics.

  1. Blogs and Social Network Sites as Activity Systems: Exploring Adult Informal Learning Process through Activity Theory Framework

    ERIC Educational Resources Information Center

    Heo, Gyeong Mi; Lee, Romee

    2013-01-01

    This paper uses an Activity Theory framework to explore adult user activities and informal learning processes as reflected in their blogs and social network sites (SNS). Using the assumption that a web-based space is an activity system in which learning occurs, typical features of the components were investigated and each activity system then…

  2. Counting Active Sites on Titanium Oxide-Silica Catalysts for Hydrogen Peroxide Activation through In Situ Poisoning with Phenylphosphonic Acid

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Eaton, Todd R.; Boston, Andrew M.; Thompson, Anthony B.

    2015-06-04

    Quantifying specific active sites in supported catalysts improves our understanding and assists in rational design. Supported oxides can undergo significant structural changes as surface densities increase from site-isolated cations to monolayers and crystallites, which changes the number of kinetically relevant sites. Herein, TiO x domains are titrated on TiO x–SiO 2 selectively with phenylphosphonic acid (PPA). An ex situ method quantifies all fluid-accessible TiO x, whereas an in situ titration during cis-cyclooctene epoxidation provides previously unavailable values for the number of tetrahedral Ti sites on which H 2O 2 activation occurs. We use this method to determine the active sitemore » densities of 22 different catalysts with different synthesis methods, loadings, and characteristic spectra and find a single intrinsic turnover frequency for cis-cyclooctene epoxidation of (40±7) h -1. This simple method gives molecular-level insight into catalyst structure that is otherwise hidden when bulk techniques are used.« less

  3. Revegetation of the riparian zone of the Three Gorges Dam Reservoir leads to increased soil bacterial diversity.

    PubMed

    Ren, Qingshui; Li, Changxiao; Yang, Wenhang; Song, Hong; Ma, Peng; Wang, Chaoying; Schneider, Rebecca L; Morreale, Stephen J

    2018-06-06

    As one of the most active components in soil, bacteria can affect soil physicochemical properties, its biological characteristics, and even its quality and health. We characterized dynamics of the soil bacterial diversity in planted (with Taxodium distichum) and unplanted soil in the riparian zone of the Three Gorges Dam Reservoir (TGDR), in southwestern China, in order to accurately quantify the changes in long-term soil bacterial community structure after revegetation. Measurements were taken annually in situ in the TGDR over the course of 5 years, from 2012 to 2016. Soil chemical properties and bacterial diversity were analyzed in both the planted and unplanted soil. After revegetation, the soil chemical properties in planted soil were significantly different than in unplanted soil. The effects of treatment, time, and the interaction of both time and treatment had significant impacts on most diversity indices. Specifically, the bacterial community diversity indices in planted soil were significantly higher and more stable than that in unplanted soil. The correlation analyses indicated that the diversity indices correlated with the pH value, organic matter, and soil available nutrients. After revegetation in the riparian zone of the TGDR, the soil quality and health is closely related to the observed bacterial diversity, and a higher bacterial diversity avails the maintenance of soil functionality. Thus, more reforestation should be carried out in the riparian zone of the TGDR, so as to effectively mitigate the negative ecological impacts of the dam. Vegetating the reservoir banks with Taxodium distichum proved successful, but planting mixed stands of native tree species could promote even higher riparian soil biodiversity and improved levels of ecosystem functioning within the TGDR.

  4. Activity-Based Probes for Isoenzyme- and Site-Specific Functional Characterization of Glutathione S -Transferases

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Stoddard, Ethan G.; Killinger, Bryan J.; Nair, Reji N.

    Glutathione S-transferases (GSTs) comprise a highly diverse family of phase II drug metabolizing enzymes whose shared function is the conjugation of reduced glutathione to various endo- and xenobiotics. Although the conglomerate activity of these enzymes can be measured by colorimetric assays, measurement of the individual contribution from specific isoforms and their contribution to the detoxification of xenobiotics in complex biological samples has not been possible. For this reason, we have developed two activity-based probes that characterize active glutathione transferases in mammalian tissues. The GST active site is comprised of a glutathione binding “G site” and a distinct substrate binding “Hmore » site”. Therefore, we developed (1) a glutathione-based photoaffinity probe (GSH-ABP) to target the “G site”, and (2) a probe designed to mimic a substrate molecule and show “H site” activity (GST-ABP). The GSH-ABP features a photoreactive moiety for UV-induced covalent binding to GSTs and glutathione-binding enzymes. The GST-ABP is a derivative of a known mechanism-based GST inhibitor that binds within the active site and inhibits GST activity. Validation of probe targets and “G” and “H” site specificity was carried out using a series of competitors in liver homogenates. Herein, we present robust tools for the novel characterization of enzyme- and active site-specific GST activity in mammalian model systems.« less

  5. Identification and characterization of the sodium-binding site of activated protein C.

    PubMed

    He, X; Rezaie, A R

    1999-02-19

    Activated protein C (APC) requires both Ca2+ and Na+ for its optimal catalytic function. In contrast to the Ca2+-binding sites, the Na+-binding site(s) of APC has not been identified. Based on a recent study with thrombin, the 221-225 loop is predicted to be a potential Na+-binding site in APC. The sequence of this loop is not conserved in trypsin. We engineered a Gla domainless form of protein C (GDPC) in which the 221-225 loop was replaced with the corresponding loop of trypsin. We found that activated GDPC (aGDPC) required Na+ (or other alkali cations) for its amidolytic activity with dissociation constant (Kd(app)) = 44.1 +/- 8.6 mM. In the presence of Ca2+, however, the requirement for Na+ by aGDPC was eliminated, and Na+ stimulated the cleavage rate 5-6-fold with Kd(app) = 2.3 +/- 0.3 mM. Both cations were required for efficient factor Va inactivation by aGDPC. In the presence of Ca2+, the catalytic function of the mutant was independent of Na+. Unlike aGDPC, the mutant did not discriminate among monovalent cations. We conclude that the 221-225 loop is a Na+-binding site in APC and that an allosteric link between the Na+ and Ca2+ binding loops modulates the structure and function of this anticoagulant enzyme.

  6. Orthogonal use of a human tRNA synthetase active site to achieve multi-functionality

    PubMed Central

    Zhou, Quansheng; Kapoor, Mili; Guo, Min; Belani, Rajesh; Xu, Xiaoling; Kiosses, William B.; Hanan, Melanie; Park, Chulho; Armour, Eva; Do, Minh-Ha; Nangle, Leslie A.; Schimmel, Paul; Yang, Xiang-Lei

    2011-01-01

    Protein multi-functionality is an emerging explanation for the complexity of higher organisms. In this regard, while aminoacyl tRNA synthetases catalyze amino acid activation for protein synthesis, some also act in pathways for inflammation, angiogenesis, and apoptosis. How multiple functions evolved and their relationship to the active site is not clear. Here structural modeling analysis, mutagenesis, and cell-based functional studies show that the potent angiostatic, natural fragment of human TrpRS associates via Trp side chains that protrude from the cognate cellular receptor VE-cadherin. Modeling indicates that (I prefer the way it was because the conclusion was reached not only by modeling, but more so by experimental studies.)VE-cadherin Trp side chains fit into the Trp-specific active site of the synthetase. Thus, specific side chains of the receptor mimic (?) amino acid substrates and expand the functionality of the active site of the synthetase. We propose that orthogonal use of the same active site may be a general way to develop multi-functionality of human tRNA synthetases and other proteins. PMID:20010843

  7. Active Layer and Moisture Measurements for Intensive Site 0 and 1, Barrow, Alaska

    DOE Data Explorer

    John Peterson

    2015-04-17

    These are measurements of Active Layer Thickness collected along several lines beginning in September, 2011 to the present. The data were collected at several time periods along the Site0 L2 Line, the Site1 AB Line, and an ERT Monitoring Line near Area A in Site1.

  8. Chronology of Miocene-Pliocene deposits at Split Mountain Gorge, Southern California: A record of regional tectonics and Colorado River evolution

    USGS Publications Warehouse

    Dorsey, R.J.; Fluette, A.; McDougall, K.; Housen, B.A.; Janecke, S.U.; Axen, G.J.; Shirvell, C.R.

    2007-01-01

    Late Miocene to early Pliocene deposit at Split Mountain Gorge, California, preserve a record of basinal response to changes in regional tectonics, paleogeography, and evolution of the Colorado River. The base of the Elephant Trees Formation, magnetostratigraphically dated as 8.1 ?? 0.4 Ma, provides the earliest well-dated record of extension in the southwestern Salton Trough. The oldest marine sediments are ca. 6.3 Ma. The nearly synchronous timing of marine incursion in the Salton Trough and northern Gulf of California region supports a model for localization of Pacific-North America plate motion in the Gulf ca. 6 Ma. The first appearance of Colorado River sand at the Miocene-Pliocene boundary (5.33 Ma) suggests rapid propagation of the river to the Salton Trough, and supports a lake-spillover hypothesis for initiation of the lower Colorado River. ?? 2007 Geological Society of America.

  9. Stereospecific suppression of active site mutants by methylphosphonate substituted substrates reveals the stereochemical course of site-specific DNA recombination

    PubMed Central

    Rowley, Paul A.; Kachroo, Aashiq H.; Ma, Chien-Hui; Maciaszek, Anna D.; Guga, Piotr; Jayaram, Makkuni

    2015-01-01

    Tyrosine site-specific recombinases, which promote one class of biologically important phosphoryl transfer reactions in DNA, exemplify active site mechanisms for stabilizing the phosphate transition state. A highly conserved arginine duo (Arg-I; Arg-II) of the recombinase active site plays a crucial role in this function. Cre and Flp recombinase mutants lacking either arginine can be rescued by compensatory charge neutralization of the scissile phosphate via methylphosphonate (MeP) modification. The chemical chirality of MeP, in conjunction with mutant recombinases, reveals the stereochemical contributions of Arg-I and Arg-II. The SP preference of the native reaction is specified primarily by Arg-I. MeP reaction supported by Arg-II is nearly bias-free or RP-biased, depending on the Arg-I substituent. Positional conservation of the arginines does not translate into strict functional conservation. Charge reversal by glutamic acid substitution at Arg-I or Arg-II has opposite effects on Cre and Flp in MeP reactions. In Flp, the base immediately 5′ to the scissile MeP strongly influences the choice between the catalytic tyrosine and water as the nucleophile for strand scission, thus between productive recombination and futile hydrolysis. The recombinase active site embodies the evolutionary optimization of interactions that not only favor the normal reaction but also proscribe antithetical side reactions. PMID:25999343

  10. Systematic Functional Analysis of Active-Site Residues in l-Threonine Dehydrogenase from Thermoplasma volcanium

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Desjardins, Morgan; Mak, Wai Shun; O’Brien, Terrence E.

    Enzymes have been through millions of years of evolution during which their active-site microenvironments are fine-tuned. Active-site residues are commonly conserved within protein families, indicating their importance for substrate recognition and catalysis. In this work, we systematically mutated active-site residues of l-threonine dehydrogenase from Thermoplasma volcanium and characterized the mutants against a panel of substrate analogs. Our results demonstrate that only a subset of these residues plays an essential role in substrate recognition and catalysis and that the native enzyme activity can be further enhanced roughly 4.6-fold by a single point mutation. Kinetic characterization of mutants on substrate analogs showsmore » that l-threonine dehydrogenase possesses promiscuous activities toward other chemically similar compounds not previously observed. Quantum chemical calculations on the hydride-donating ability of these substrates also reveal that this enzyme did not evolve to harness the intrinsic substrate reactivity for enzyme catalysis. Our analysis provides insights into connections between the details of enzyme active-site structure and specific function. Finally, these results are directly applicable to rational enzyme design and engineering.« less

  11. Systematic Functional Analysis of Active-Site Residues in l-Threonine Dehydrogenase from Thermoplasma volcanium

    DOE PAGES

    Desjardins, Morgan; Mak, Wai Shun; O’Brien, Terrence E.; ...

    2017-07-07

    Enzymes have been through millions of years of evolution during which their active-site microenvironments are fine-tuned. Active-site residues are commonly conserved within protein families, indicating their importance for substrate recognition and catalysis. In this work, we systematically mutated active-site residues of l-threonine dehydrogenase from Thermoplasma volcanium and characterized the mutants against a panel of substrate analogs. Our results demonstrate that only a subset of these residues plays an essential role in substrate recognition and catalysis and that the native enzyme activity can be further enhanced roughly 4.6-fold by a single point mutation. Kinetic characterization of mutants on substrate analogs showsmore » that l-threonine dehydrogenase possesses promiscuous activities toward other chemically similar compounds not previously observed. Quantum chemical calculations on the hydride-donating ability of these substrates also reveal that this enzyme did not evolve to harness the intrinsic substrate reactivity for enzyme catalysis. Our analysis provides insights into connections between the details of enzyme active-site structure and specific function. Finally, these results are directly applicable to rational enzyme design and engineering.« less

  12. Active site of tripeptidyl peptidase II from human erythrocytes is of the subtilisin type

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tomkinson, B.; Wernstedt, C.; Hellman, U.

    1987-11-01

    The present report presents evidence that the amino acid sequence around the serine of the active site of human tripeptidyl peptidase II is of the subtilisin type. The enzyme from human erythrocytes was covalently labeled at its active site with (/sup 3/H)diisopropyl fluorophosphate, and the protein was subsequently reduced, alkylated, and digested with trypsin. The labeled tryptic peptides were purified by gel filtration and repeated reversed-phase HPLC, and their amino-terminal sequences were determined. Residue 9 contained the radioactive label and was, therefore, considered to be the active serine residue. The primary structure of the part of the active site (residuesmore » 1-10) containing this residue was concluded to be Xaa-Thr-Gln-Leu-Met-Asx-Gly-Thr-Ser-Met. This amino acid sequence is homologous to the sequence surrounding the active serine of the microbial peptidases subtilisin and thermitase. These data demonstrate that human tripeptidyl peptidase II represents a potentially distinct class of human peptidases and raise the question of an evolutionary relationship between the active site of a mammalian peptidase and that of the subtilisin family of serine peptidases.« less

  13. Active site of tripeptidyl peptidase II from human erythrocytes is of the subtilisin type.

    PubMed Central

    Tomkinson, B; Wernstedt, C; Hellman, U; Zetterqvist, O

    1987-01-01

    The present report presents evidence that the amino acid sequence around the serine of the active site of human tripeptidyl peptidase II is of the subtilisin type. The enzyme from human erythrocytes was covalently labeled at its active site with [3H]diisopropyl fluorophosphate, and the protein was subsequently reduced, alkylated, and digested with trypsin. The labeled tryptic peptides were purified by gel filtration and repeated reversed-phase HPLC, and their amino-terminal sequences were determined. Residue 9 contained the radioactive label and was, therefore, considered to be the active serine residue. The primary structure of the part of the active site (residues 1-10) containing this residue was concluded to be Xaa-Thr-Gln-Leu-Met-Asx-Gly-Thr-Ser-Met. This amino acid sequence is homologous to the sequence surrounding the active serine of the microbial peptidases subtilisin and thermitase. These data demonstrate that human tripeptidyl peptidase II represents a potentially distinct class of human peptidases and raise the question of an evolutionary relationship between the active site of a mammalian peptidase and that of the subtilisin family of serine peptidases. PMID:3313395

  14. Relationship between the transmission of schistosomiasis japonica and the construction of the Three Gorge Reservoir.

    PubMed

    Zheng, Jiang; Gu, Xue-guang; Xu, Yong-long; Ge, Ji-hua; Yang, Xian-xiang; He, Chang-hao; Tang, Chao; Cai, Kai-ping; Jiang, Qing-wu; Liang, You-sheng; Wang, Tian-ping; Xu, Xing-jian; Zhong, Jiu-he; Yuan, Hong-chang; Zhou, Xiao-nong

    2002-05-01

    To study the relationship between the changes of environment and the transmission of Schistosomiasis japonica after the construction of the Three Gorge Reservoir. On the basis of the predictive data on changes of water level and sediment in the middle and lower reaches of the Yangtze River after the dam construction provided by the Yangtze River Committee on Water Conservancy, corresponding data were collected and analyzed. In order to understand the effects of changes of water level and sediment on Oncomelania hupensis, the intermediate host of Schistosoma japonicum, field surveys and ecologically intimated experiments were done. The prevalence status and factors related to transmission were investigated by epidemiology, medical geography and public health. Positive impact on control of schistosomiasis is that floods in the Yangtze River will occur less and will decrease the dispersal of snails and the chances of infection for humans and animals as well. Snail habitats' beach will be reduced in the flushed area along the Yangtze River. Exploitation of beach and elimination of snails will benefit from the beach's uplift in flushed area. The floods ahead of time in autumn will interfere with the development of snails. Negative impact is that the flushed beaches and migratory settlements at certain altitude may become snail habitats. So the reservoir area will be a potential transmission area of schistosomiasis due to dispersal of infectious resources and Oncomelania snails. The uprising of the water body level may cause the expending of the snail habitats in some places of Jianghan plain during the transition of spring to summer. The destruction of the embankments of the Dongting Lake will be put off by its delayed flush, which causes the beach in a situation of growing reed-grass or grass for a long time that benefits the reproduction of the snails. The chances for schistosome infection to humans and animals will be on the increase, because the water regression

  15. Calorimetric studies of the interactions of metalloenzyme active site mimetics with zinc-binding inhibitors.

    PubMed

    Robinson, Sophia G; Burns, Philip T; Miceli, Amanda M; Grice, Kyle A; Karver, Caitlin E; Jin, Lihua

    2016-07-19

    The binding of drugs to metalloenzymes is an intricate process that involves several interactions, including binding of the drug to the enzyme active site metal, as well as multiple interactions between the drug and the enzyme residues. In order to determine the free energy contribution of Zn(2+) binding by known metalloenzyme inhibitors without the other interactions, valid active site zinc structural mimetics must be formed and binding studies need to be performed in biologically relevant conditions. The potential of each of five ligands to form a structural mimetic with Zn(2+) was investigated in buffer using Isothermal Titration Calorimetry (ITC). All five ligands formed strong 1 : 1 (ligand : Zn(2+)) binary complexes. The complexes were used in further ITC experiments to study their interaction with 8-hydroxyquinoline (8-HQ) and/or acetohydroxamic acid (AHA), two bidentate anionic zinc-chelating enzyme inhibitors. It was found that tetradentate ligands were not suitable for creating zinc structural mimetics for inhibitor binding in solution due to insufficient coordination sites remaining on Zn(2+). A stable binary complex, [Zn(BPA)](2+), which was formed by a tridentate ligand, bis(2-pyridylmethyl)amine (BPA), was found to bind one AHA in buffer or a methanol : buffer mixture (60 : 40 by volume) at pH 7.25 or one 8-HQ in the methanol : buffer mixture at pH 6.80, making it an effective structural mimetic for the active site of zinc metalloenzymes. These results are consistent with the observation that metalloenzyme active site zinc ions have three residues coordinated to them, leaving one or two sites open for inhibitors to bind. Our findings indicate that Zn(BPA)X2 can be used as an active site structural mimetic for zinc metalloenzymes for estimating the free energy contribution of zinc binding to the overall inhibitor active site interactions. Such use will help aid in the rational design of inhibitors to a variety of zinc metalloenzymes.

  16. Active site electrostatics protect genome integrity by blocking abortive hydrolysis during DNA recombination

    PubMed Central

    Ma, Chien-Hui; Rowley, Paul A; Macieszak, Anna; Guga, Piotr; Jayaram, Makkuni

    2009-01-01

    Water, acting as a rogue nucleophile, can disrupt transesterification steps of important phosphoryl transfer reactions in DNA and RNA. We have unveiled this risk, and identified safeguards instituted against it, during strand cleavage and joining by the tyrosine site-specific recombinase Flp. Strand joining is threatened by a latent Flp endonuclease activity (type I) towards the 3′-phosphotyrosyl intermediate resulting from strand cleavage. This risk is not alleviated by phosphate electrostatics; neutralizing the negative charge on the scissile phosphate through methylphosphonate (MeP) substitution does not stimulate type I endonuclease. Rather, protection derives from the architecture of the recombination synapse and conformational dynamics within it. Strand cleavage is protected against water by active site electrostatics. Replacement of the catalytic Arg-308 of Flp by alanine, along with MeP substitution, elicits a second Flp endonuclease activity (type II) that directly targets the scissile phosphodiester bond in DNA. MeP substitution, combined with appropriate active site mutations, will be useful in revealing anti-hydrolytic mechanisms engendered by systems that mediate DNA relaxation, DNA transposition, site-specific recombination, telomere resolution, RNA splicing and retrohoming of mobile introns. PMID:19440204

  17. Expansion of access tunnels and active-site cavities influence activity of haloalkane dehalogenases in organic cosolvents.

    PubMed

    Stepankova, Veronika; Khabiri, Morteza; Brezovsky, Jan; Pavelka, Antonin; Sykora, Jan; Amaro, Mariana; Minofar, Babak; Prokop, Zbynek; Hof, Martin; Ettrich, Rudiger; Chaloupkova, Radka; Damborsky, Jiri

    2013-05-10

    The use of enzymes for biocatalysis can be significantly enhanced by using organic cosolvents in the reaction mixtures. Selection of the cosolvent type and concentration range for an enzymatic reaction is challenging and requires extensive empirical testing. An understanding of protein-solvent interaction could provide a theoretical framework for rationalising the selection process. Here, the behaviour of three model enzymes (haloalkane dehalogenases) was investigated in the presence of three representative organic cosolvents (acetone, formamide, and isopropanol). Steady-state kinetics assays, molecular dynamics simulations, and time-resolved fluorescence spectroscopy were used to elucidate the molecular mechanisms of enzyme-solvent interactions. Cosolvent molecules entered the enzymes' access tunnels and active sites, enlarged their volumes with no change in overall protein structure, but surprisingly did not act as competitive inhibitors. At low concentrations, the cosolvents either enhanced catalysis by lowering K(0.5) and increasing k(cat), or caused enzyme inactivation by promoting substrate inhibition and decreasing k(cat). The induced activation and inhibition of the enzymes correlated with expansion of the active-site pockets and their occupancy by cosolvent molecules. The study demonstrates that quantitative analysis of the proportions of the access tunnels and active-sites occupied by organic solvent molecules provides the valuable information for rational selection of appropriate protein-solvent pair and effective cosolvent concentration. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Using Carbohydrate Interaction Assays to Reveal Novel Binding Sites in Carbohydrate Active Enzymes.

    PubMed

    Cockburn, Darrell; Wilkens, Casper; Dilokpimol, Adiphol; Nakai, Hiroyuki; Lewińska, Anna; Abou Hachem, Maher; Svensson, Birte

    2016-01-01

    Carbohydrate active enzymes often contain auxiliary binding sites located either on independent domains termed carbohydrate binding modules (CBMs) or as so-called surface binding sites (SBSs) on the catalytic module at a certain distance from the active site. The SBSs are usually critical for the activity of their cognate enzyme, though they are not readily detected in the sequence of a protein, but normally require a crystal structure of a complex for their identification. A variety of methods, including affinity electrophoresis (AE), insoluble polysaccharide pulldown (IPP) and surface plasmon resonance (SPR) have been used to study auxiliary binding sites. These techniques are complementary as AE allows monitoring of binding to soluble polysaccharides, IPP to insoluble polysaccharides and SPR to oligosaccharides. Here we show that these methods are useful not only for analyzing known binding sites, but also for identifying new ones, even without structural data available. We further verify the chosen assays discriminate between known SBS/CBM containing enzymes and negative controls. Altogether 35 enzymes are screened for the presence of SBSs or CBMs and several novel binding sites are identified, including the first SBS ever reported in a cellulase. This work demonstrates that combinations of these methods can be used as a part of routine enzyme characterization to identify new binding sites and advance the study of SBSs and CBMs, allowing them to be detected in the absence of structural data.

  19. Using Carbohydrate Interaction Assays to Reveal Novel Binding Sites in Carbohydrate Active Enzymes

    PubMed Central

    Wilkens, Casper; Dilokpimol, Adiphol; Nakai, Hiroyuki; Lewińska, Anna; Abou Hachem, Maher; Svensson, Birte

    2016-01-01

    Carbohydrate active enzymes often contain auxiliary binding sites located either on independent domains termed carbohydrate binding modules (CBMs) or as so-called surface binding sites (SBSs) on the catalytic module at a certain distance from the active site. The SBSs are usually critical for the activity of their cognate enzyme, though they are not readily detected in the sequence of a protein, but normally require a crystal structure of a complex for their identification. A variety of methods, including affinity electrophoresis (AE), insoluble polysaccharide pulldown (IPP) and surface plasmon resonance (SPR) have been used to study auxiliary binding sites. These techniques are complementary as AE allows monitoring of binding to soluble polysaccharides, IPP to insoluble polysaccharides and SPR to oligosaccharides. Here we show that these methods are useful not only for analyzing known binding sites, but also for identifying new ones, even without structural data available. We further verify the chosen assays discriminate between known SBS/CBM containing enzymes and negative controls. Altogether 35 enzymes are screened for the presence of SBSs or CBMs and several novel binding sites are identified, including the first SBS ever reported in a cellulase. This work demonstrates that combinations of these methods can be used as a part of routine enzyme characterization to identify new binding sites and advance the study of SBSs and CBMs, allowing them to be detected in the absence of structural data. PMID:27504624

  20. Active-site protein dynamics and solvent accessibility in native Achromobacter cycloclastes copper nitrite reductase.

    PubMed

    Sen, Kakali; Horrell, Sam; Kekilli, Demet; Yong, Chin W; Keal, Thomas W; Atakisi, Hakan; Moreau, David W; Thorne, Robert E; Hough, Michael A; Strange, Richard W

    2017-07-01

    Microbial nitrite reductases are denitrifying enzymes that are a major component of the global nitrogen cycle. Multiple structures measured from one crystal (MSOX data) of copper nitrite reductase at 240 K, together with molecular-dynamics simulations, have revealed protein dynamics at the type 2 copper site that are significant for its catalytic properties and for the entry and exit of solvent or ligands to and from the active site. Molecular-dynamics simulations were performed using different protonation states of the key catalytic residues (Asp CAT and His CAT ) involved in the nitrite-reduction mechanism of this enzyme. Taken together, the crystal structures and simulations show that the Asp CAT protonation state strongly influences the active-site solvent accessibility, while the dynamics of the active-site 'capping residue' (Ile CAT ), a determinant of ligand binding, are influenced both by temperature and by the protonation state of Asp CAT . A previously unobserved conformation of Ile CAT is seen in the elevated temperature series compared with 100 K structures. DFT calculations also show that the loss of a bound water ligand at the active site during the MSOX series is consistent with reduction of the type 2 Cu atom.

  1. An active site-tail interaction in the structure of hexahistidine-tagged Thermoplasma acidophilum citrate synthase

    DOE PAGES

    Murphy, Jesse R.; Donini, Stefano; Kappock, T. Joseph

    2015-10-01

    Citrate synthase (CS) plays a central metabolic role in aerobes and many other organisms. The CS reaction comprises two half-reactions: a Claisen aldol condensation of acetyl-CoA (AcCoA) and oxaloacetate (OAA) that forms citryl-CoA (CitCoA), and CitCoA hydrolysis. Protein conformational changes that `close' the active site play an important role in the assembly of a catalytically competent condensation active site. CS from the thermoacidophile Thermoplasma acidophilum (TpCS) possesses an endogenous Trp fluorophore that can be used to monitor the condensation reaction. The 2.2 Å resolution crystal structure of TpCS fused to a C-terminal hexahistidine tag (TpCSH6) reported here is an `open'more » structure that, when compared with several liganded TpCS structures, helps to define a complete path for active-site closure. One active site in each dimer binds a neighboring His tag, the first nonsubstrate ligand known to occupy both the AcCoA and OAA binding sites. Solution data collectively suggest that this fortuitous interaction is stabilized by the crystalline lattice. In conclusion, as a polar but almost neutral ligand, the active site-tail interaction provides a new starting point for the design of bisubstrate-analog inhibitors of CS.« less

  2. An active site-tail interaction in the structure of hexahistidine-tagged Thermoplasma acidophilum citrate synthase.

    PubMed

    Murphy, Jesse R; Donini, Stefano; Kappock, T Joseph

    2015-10-01

    Citrate synthase (CS) plays a central metabolic role in aerobes and many other organisms. The CS reaction comprises two half-reactions: a Claisen aldol condensation of acetyl-CoA (AcCoA) and oxaloacetate (OAA) that forms citryl-CoA (CitCoA), and CitCoA hydrolysis. Protein conformational changes that `close' the active site play an important role in the assembly of a catalytically competent condensation active site. CS from the thermoacidophile Thermoplasma acidophilum (TpCS) possesses an endogenous Trp fluorophore that can be used to monitor the condensation reaction. The 2.2 Å resolution crystal structure of TpCS fused to a C-terminal hexahistidine tag (TpCSH6) reported here is an `open' structure that, when compared with several liganded TpCS structures, helps to define a complete path for active-site closure. One active site in each dimer binds a neighboring His tag, the first nonsubstrate ligand known to occupy both the AcCoA and OAA binding sites. Solution data collectively suggest that this fortuitous interaction is stabilized by the crystalline lattice. As a polar but almost neutral ligand, the active site-tail interaction provides a new starting point for the design of bisubstrate-analog inhibitors of CS.

  3. A simplified method for active-site titration of lipases immobilised on hydrophobic supports.

    PubMed

    Nalder, Tim D; Kurtovic, Ivan; Barrow, Colin J; Marshall, Susan N

    2018-06-01

    The aim of this work was to develop a simple and accurate protocol to measure the functional active site concentration of lipases immobilised on highly hydrophobic supports. We used the potent lipase inhibitor methyl 4-methylumbelliferyl hexylphosphonate to titrate the active sites of Candida rugosa lipase (CrL) bound to three highly hydrophobic supports: octadecyl methacrylate (C18), divinylbenzene crosslinked methacrylate (DVB) and styrene. The method uses correction curves to take into account the binding of the fluorophore (4-methylumbelliferone, 4-MU) by the support materials. We showed that the uptake of the detection agent by the three supports is not linear relative to the weight of the resin, and that the uptake occurs in an equilibrium that is independent of the total fluorophore concentration. Furthermore, the percentage of bound fluorophore varied among the supports, with 50 mg of C18 and styrene resins binding approximately 64 and 94%, respectively. When the uptake of 4-MU was calculated and corrected for, the total 4-MU released via inhibition (i.e. the concentration of functional lipase active sites) could be determined via a linear relationship between immobilised lipase weight and total inhibition. It was found that the functional active site concentration of immobilised CrL varied greatly among different hydrophobic supports, with 56% for C18, compared with 14% for DVB. The described method is a simple and robust approach to measuring functional active site concentration in immobilised lipase samples. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. Non-canonical active site architecture of the radical SAM thiamin pyrimidine synthase.

    PubMed

    Fenwick, Michael K; Mehta, Angad P; Zhang, Yang; Abdelwahed, Sameh H; Begley, Tadhg P; Ealick, Steven E

    2015-03-27

    Radical S-adenosylmethionine (SAM) enzymes use a [4Fe-4S] cluster to generate a 5'-deoxyadenosyl radical. Canonical radical SAM enzymes are characterized by a β-barrel-like fold and SAM anchors to the differentiated iron of the cluster, which is located near the amino terminus and within the β-barrel, through its amino and carboxylate groups. Here we show that ThiC, the thiamin pyrimidine synthase in plants and bacteria, contains a tethered cluster-binding domain at its carboxy terminus that moves in and out of the active site during catalysis. In contrast to canonical radical SAM enzymes, we predict that SAM anchors to an additional active site metal through its amino and carboxylate groups. Superimposition of the catalytic domains of ThiC and glutamate mutase shows that these two enzymes share similar active site architectures, thus providing strong evidence for an evolutionary link between the radical SAM and adenosylcobalamin-dependent enzyme superfamilies.

  5. [Nuclease activity of the recombinant plancitoxin-1-like proteins with mutations in the active site from Trichinella spiralis].

    PubMed

    Liao, Chengshui; Wang, Xiaoli; Tian, Wenjing; Zhang, Mengke; Zhang, Chunjie; Li, Yinju; Wu, Tingcai; Cheng, Xiangchao

    2017-08-25

    Although there are 125 predicted DNase Ⅱ-like family genes in the Trichinella spiralis genome, plancitoxin-1-like (Ts-Pt) contains the HKD motif, a typical conserved region of DNase Ⅱ, in N- and C-terminal. It is generally believed that histidine is the active site in DNase Ⅱ. To study the nuclease activity of recombinant Ts-Pt with mutations in the active site from T. spiralis, different fragments of the mutated Ts-Pt genes were cloned using overlap PCR technique and inserted into the expressing vector pET-28a(+), and transformed into Escherichia coli Rosseta (DE3). The fusion proteins were purified by Ni-NTA affinity chromatography and SDS-PAGE. Nuclease activity of the recombinant proteins was detected by agarose gel electrophoresis and nuclease-zymography. The recombinant plasmids harboring the mutated Ts-Pt genes were constructed and expressed as inclusive body in a prokaryotic expression system. After renaturation in vitro, the recombinant proteins had no nuclease activity according to agarose gel electrophoresis. However, the expressed proteins as inclusive body displayed the ability to degrade DNA after renaturation in gel. And the nuclease activity was not affected after subjected to mutation of active site in N- and C-termini of Ts-Pt. These results provide the basis to study the relationship between DNase Ⅱ-like protein family and infection of T. spiralis.

  6. A Variable Active Site Residue Influences the Kinetics of Response Regulator Phosphorylation and Dephosphorylation.

    PubMed

    Immormino, Robert M; Silversmith, Ruth E; Bourret, Robert B

    2016-10-04

    Two-component regulatory systems, minimally composed of a sensor kinase and a response regulator protein, are common mediators of signal transduction in microorganisms. All response regulators contain a receiver domain with conserved active site residues that catalyze the signal activating and deactivating phosphorylation and dephosphorylation reactions. We explored the impact of variable active site position T+1 (one residue C-terminal to the conserved Thr/Ser) on reaction kinetics and signaling fidelity, using wild type and mutant Escherichia coli CheY, CheB, and NarL to represent the three major sequence classes observed across response regulators: Ala/Gly, Ser/Thr, and Val/Ile/Met, respectively, at T+1. Biochemical and structural data together suggested that different amino acids at T+1 impacted reaction kinetics by altering access to the active site while not perturbing overall protein structure. A given amino acid at position T+1 had similar effects on autodephosphorylation in each protein background tested, likely by modulating access of the attacking water molecule to the active site. Similarly, rate constants for CheY autophosphorylation with three different small molecule phosphodonors were consistent with the steric constraints on access to the phosphorylation site arising from combination of specific phosphodonors with particular amino acids at T+1. Because other variable active site residues also influence response regulator phosphorylation biochemistry, we began to explore how context (here, the amino acid at T+2) affected the influence of position T+1 on CheY autocatalytic reactions. Finally, position T+1 affected the fidelity and kinetics of phosphotransfer between sensor kinases and response regulators but was not a primary determinant of their interaction.

  7. Insights into the glycyl radical enzyme active site of benzylsuccinate synthase: a computational study.

    PubMed

    Bharadwaj, Vivek S; Dean, Anthony M; Maupin, C Mark

    2013-08-21

    The fumarate addition reaction, catalyzed by the enzyme benzylsuccinate synthase (BSS), is considered to be one of the most intriguing and energetically challenging reactions in biology. BSS belongs to the glycyl radical enzyme family and catalyzes the fumarate addition reaction, which enables microorganisms to utilize hydrocarbons as an energy source under anaerobic conditions. Unfortunately, the extreme sensitivity of the glycyl radical to oxygen has hampered the structural and kinetic characterization of BSS, thereby limiting our knowledge on this enzyme. To enhance our molecular-level understanding of BSS, a computational approach involving homology modeling, docking studies, and molecular dynamics (MD) simulations has been used to deduce the structure of BSS's catalytic subunit (BSSα) and illuminate the molecular basis for the fumarate addition reaction. We have identified two conserved and distinct binding pockets at the BSSα active site: a hydrophobic pocket for toluene binding and a polar pocket for fumaric acid binding. Subsequent dynamical and energetic evaluations have identified Glu509, Ser827, Leu390, and Phe384 as active site residues critical for substrate binding. The orientation of substrates at the active site observed in MD simulations is consistent with experimental observations of the syn addition of toluene to fumaric acid. It is also found that substrate binding tightens the active site and restricts the conformational flexibility of the thiyl radical, leading to hydrogen transfer distances conducive to the proposed reaction mechanism. The stability of substrates at the active site and the occurrence of feasible radical transfer distances between the thiyl radical, substrates, and the active site glycine indicate a substrate-assisted radical transfer pathway governing fumarate addition.

  8. Flexibility and Stability Trade-Off in Active Site of Cold-Adapted Pseudomonas mandelii Esterase EstK.

    PubMed

    Truongvan, Ngoc; Jang, Sei-Heon; Lee, ChangWoo

    2016-06-28

    Cold-adapted enzymes exhibit enhanced conformational flexibility, especially in their active sites, as compared with their warmer-temperature counterparts. However, the mechanism by which cold-adapted enzymes maintain their active site stability is largely unknown. In this study, we investigated the role of conserved D308-Y309 residues located in the same loop as the catalytic H307 residue in the cold-adapted esterase EstK from Pseudomonas mandelii. Mutation of D308 and/or Y309 to Ala or deletion resulted in increased conformational flexibility. Particularly, the D308A or Y309A mutant showed enhanced substrate affinity and catalytic rate, as compared with wild-type EstK, via enlargement of the active site. However, all mutant EstK enzymes exhibited reduced thermal stability. The effect of mutation was greater for D308 than Y309. These results indicate that D308 is not preferable for substrate selection and catalytic activity, whereas hydrogen bond formation involving D308 is critical for active site stabilization. Taken together, conformation of the EstK active site is constrained via flexibility-stability trade-off for enzyme catalysis and thermal stability. Our study provides further insights into active site stabilization of cold-adapted enzymes.

  9. Well-Defined Metal-O6 in Metal-Catecholates as a Novel Active Site for Oxygen Electroreduction.

    PubMed

    Liu, Xuan-He; Hu, Wei-Li; Jiang, Wen-Jie; Yang, Ya-Wen; Niu, Shuai; Sun, Bing; Wu, Jing; Hu, Jin-Song

    2017-08-30

    Metal-nitrogen coordination sites, M-N x (M = Fe, Co, Ni, etc.), have shown great potential to replace platinum group materials as electrocatalysts for oxygen reduction reaction (ORR). However, the real active site in M-N x is still vague to date due to their complicated structure and composition. It is therefore highly desirable but challenging to develop ORR catalysts with novel and clear active sites, which could meet the needs of comprehensive understanding of structure-function relationships and explore new cost-effective and efficient ORR electrocatalysts. Herein, well-defined M-O 6 coordination in metal-catecholates (M-CATs, M = Ni or Co) is discovered to be catalytically active for ORR via a four-electron-dominated pathway. In view of no pyrolysis involved and unambiguous crystalline structure of M-CATs, the M-O 6 octahedral coordination site with distinct structure is determined as a new type of active site for ORR. These findings extend the scope of metal-nonmetal coordination as an active site for ORR and pave a way for bottom-up design of novel electrocatalysts containing M-O 6 coordination.

  10. The structure of amylosucrase from Deinococcus radiodurans has an unusual open active-site topology.

    PubMed

    Skov, Lars K; Pizzut-Serin, Sandra; Remaud-Simeon, Magali; Ernst, Heidi A; Gajhede, Michael; Mirza, Osman

    2013-09-01

    Amylosucrases (ASes) catalyze the formation of an α-1,4-glucosidic linkage by transferring a glucosyl unit from sucrose onto an acceptor α-1,4-glucan. To date, several ligand-bound crystal structures of wild-type and mutant ASes from Neisseria polysaccharea and Deinococcus geothermalis have been solved. These structures all display a very similar overall conformation with a deep pocket leading to the site for transglucosylation, subsite -1. This has led to speculation on how sucrose enters the active site during glucan elongation. In contrast to previous studies, the AS structure from D. radiodurans presented here has a completely empty -1 subsite. This structure is strikingly different from other AS structures, as an active-site-lining loop comprising residues Leu214-Asn225 is found in a previously unobserved conformation. In addition, a large loop harbouring the conserved active-site residues Asp133 and Tyr136 is disordered. The result of the changed loop conformations is that the active-site topology is radically changed, leaving subsite -1 exposed and partially dismantled. This structure provides novel insights into the dynamics of ASes and comprises the first structural support for an elongation mechanism that involves considerable conformational changes to modulate accessibility to the sucrose-binding site and thereby allows successive cycles of glucosyl-moiety transfer to a growing glucan chain.

  11. Active chemisorption sites in functionalized ionic liquids for carbon capture.

    PubMed

    Cui, Guokai; Wang, Jianji; Zhang, Suojiang

    2016-07-25

    Development of novel technologies for the efficient and reversible capture of CO2 is highly desired. In the last decade, CO2 capture using ionic liquids has attracted intensive attention from both academia and industry, and has been recognized as a very promising technology. Recently, a new approach has been developed for highly efficient capture of CO2 by site-containing ionic liquids through chemical interaction. This perspective review focuses on the recent advances in the chemical absorption of CO2 using site-containing ionic liquids, such as amino-based ionic liquids, azolate ionic liquids, phenolate ionic liquids, dual-functionalized ionic liquids, pyridine-containing ionic liquids and so on. Other site-containing liquid absorbents such as amine-based solutions, switchable solvents, and functionalized ionic liquid-amine blends are also investigated. Strategies have been discussed for how to activate the existent reactive sites and develop novel reactive sites by physical and chemical methods to enhance CO2 absorption capacity and reduce absorption enthalpy. The carbon capture mechanisms of these site-containing liquid absorbents are also presented. Particular attention has been paid to the latest progress in CO2 capture in multiple-site interactions by amino-free anion-functionalized ionic liquids. In the last section, future directions and prospects for carbon capture by site-containing ionic liquids are outlined.

  12. Disturbance of Dabao highway construction on plant species and soil nutrients in Longitudinal Range Gorge Region (LRGR) of Southwestern China.

    PubMed

    Cui, Baoshan; Zhao, Shuqing; Zhang, Kejiang; Li, Shaocai; Dong, Shikui; Bai, Junhong

    2009-11-01

    The disturbance of highway construction upon surrounding vulnerable ecosystems is a common threat in the Longitudinal Range Gorge Region of southwestern China. We evaluated the disturbance of highway on plant species richness and diversity and soil nutrients from adjacent to the highway to 300 m upslope and 100 m downslope in forests and grasslands by setting 12 belt transects in forests and grasslands (six belt transects and six control belt transects, respectively). The results showed that there were some significant variances in belt transects with respective control belt transects for species richness and diversity in both forests and grasslands. Species richness and diversity of trees were lower within a 50-m distance from the highway and more noticeable on the downslope portion. Species richness and diversity of shrubs and herbs appeared higher near highway edge. Both species richness and diversity of herbs were similar in forests. In addition, exotic species, such as Eupatorium adenophorum, were further from the road and more widely dispersed in grasslands. Soil nutrients except total potassium (TK) were lower in the downslope area adjacent to highway edge and showed a significant increase with increasing distance from the highway in both forests and grasslands. This indicates that grasslands acted as microhabitats for exotic species and are more easily to be invaded than forests, especially if disturbed. Once destroyed, plant species and soil nutrients will require a significant amount of time to be restored to control levels. This work illustrates that the effects extend considerably to distances upslope and downslope from the construction site. Given that these changes occurred relatively quickly, the study suggests that the environmental "footprint" grows far beyond the road and adjacent zone of disruption.

  13. Potential effects of four Flaming Gorge Dam hydropower operational scenarios on the fishes of the Green River, Utah and Colorado

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hlohowskyj, I.; Hayse, J.W.

    1995-09-01

    Aerial videography and modeling were used to evaluate the impacts of four hydropower operational scenarios at Flaming Gorge Dam, Utah, on trout and native fishes in the Green River, Utah and Colorado. The four operational scenarios studied were year-round high fluctuations, seasonally adjusted high fluctuations, seasonally adjusted moderate fluctuations, and seasonally adjusted steady flows. Impacts on trout were evaluated by examining differences among scenarios in the areas of inundated substrates that serve as spawning and feeding habitat. All scenarios would provide at least 23 acres per mile of habitat for spawning and food production; seasonally adjusted operations would provide additionalmore » areas during periods of sustained high release. Seasonally adjusted high fluctuations would increase inundated areas by 12 to 26% for a short period in winter and spring, but food production and reproduction would not be expected to increase. Seasonally adjusted moderate fluctuations and steady flows would produce similar increases in area, but the longer period of inundation could also result in increased food production and provide additional spawning sites for trout. Impacts on native fishes were assessed by examining daily changes in backwater nursery areas. Compared with year-round high fluctuations, the daily changes in backwater area would decrease by about 47, 89, and 100% under the seasonally adjusted high fluctuation, moderate fluctuation, and steady flow scenarios, respectively. Similarly, daily stage fluctuations during the nursery period would decrease by 72, 89, and 100% under the seasonally adjusted high fluctuation, moderate fluctuation, and steady flow scenarios, respectively. These reductions in daily fluctuations in backwater area and stage would improve conditions in nursery habitats and could in turn improve recruitment and overwinter survival. Introduced fish species could also benefit from the seasonally adjusted operational scenarios.« less

  14. Structural evolution of luciferase activity in Zophobas mealworm AMP/CoA-ligase (protoluciferase) through site-directed mutagenesis of the luciferin binding site.

    PubMed

    Prado, R A; Barbosa, J A; Ohmiya, Y; Viviani, V R

    2011-07-01

    The structural origin and evolution of bioluminescent activity of beetle luciferases from AMP/CoA ligases remains a mystery. Previously we cloned the luciferase-like enzyme from Zophobas morio mealworm, a reasonable protoluciferase model that could shine light on this mystery. Kinetic characterization and studies with D- and L-luciferin and their adenylates showed that stereoselectivity constitutes a critical feature for the origin of luciferase activity in AMP/CoA ligases. Comparison of the primary structures and modeling studies of this protoluciferase and the three main families of beetle luciferases showed that the carboxylic acid substrate binding site of this enzyme is smaller and more hydrophobic than the luciferin binding site of beetle luciferases, showing several substitutions of otherwise conserved residues. Thus, here we performed a site-directed mutagenesis survey of the carboxylic binding site motifs of the protoluciferase by replacing their residues by the respective conserved ones found in beetle luciferases in order to identify the structural determinants of luciferase/oxygenase activity. Although most of the substitutions had negative impact on the luminescence activity of the protoluciferase, only the substitution I327T improved the luminescence activity, resulting in a broad and 15 nm blue-shifted luminescence spectrum. Such substitution indicates the importance of the loop motif 322YGMSEI327 (341YGLTETT347 in Photinus pyralis luciferase) for luciferase activity, and indicates a possible route for the evolution of bioluminescence function of beetle luciferases.

  15. Probing the Active Surface Sites for CO Reduction on Oxide-Derived Copper Electrocatalysts

    DOE PAGES

    Verdaguer-Casadevall, Arnau; Li, Christina W.; Johansson, Tobias P.; ...

    2015-07-30

    CO electroreduction activity on oxide-derived Cu (OD-Cu) was found to correlate with metastable surface features that bind CO strongly. OD-Cu electrodes prepared by H 2 reduction of Cu 2O precursors reduce CO to acetate and ethanol with nearly 50% Faradaic efficiency at moderate overpotential. Temperature-programmed desorption of CO on OD-Cu revealed the presence of surface sites with strong CO binding that are distinct from the terraces and stepped sites found on polycrystalline Cu foil. After annealing at 350 °C, the surface-area corrected current density for CO reduction is 44-fold lower and the Faradaic efficiency is less than 5%. These changesmore » are accompanied by a reduction in the proportion of strong CO binding sites. Here, we propose that the active sites for CO reduction on OD-Cu surfaces are strong CO binding sites that are supported by grain boundaries. Uncovering these sites is a first step toward understanding the surface chemistry necessary for efficient CO electroreduction.« less

  16. Substrate Shuttling Between Active Sites of Uroporphyrinogen Decarboxylase in Not Required to Generate Coproporphyrinogen

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Phillips, J.; Warby, C; Whitby, F

    2009-01-01

    Uroporphyrinogen decarboxylase (URO-D; EC 4.1.1.37), the fifth enzyme of the heme biosynthetic pathway, is required for the production of heme, vitamin B12, siroheme, and chlorophyll precursors. URO-D catalyzes the sequential decarboxylation of four acetate side chains in the pyrrole groups of uroporphyrinogen to produce coproporphyrinogen. URO-D is a stable homodimer, with the active-site clefts of the two subunits adjacent to each other. It has been hypothesized that the two catalytic centers interact functionally, perhaps by shuttling of reaction intermediates between subunits. We tested this hypothesis by construction of a single-chain protein (single-chain URO-D) in which the two subunits were connectedmore » by a flexible linker. The crystal structure of this protein was shown to be superimposable with wild-type activity and to have comparable catalytic activity. Mutations that impaired one or the other of the two active sites of single-chain URO-D resulted in approximately half of wild-type activity. The distributions of reaction intermediates were the same for mutant and wild-type sequences and were unaltered in a competition experiment using I and III isomer substrates. These observations indicate that communication between active sites is not required for enzyme function and suggest that the dimeric structure of URO-D is required to achieve conformational stability and to create a large active-site cleft.« less

  17. Quantifying the impact of the Three Gorges Dam on the thermal dynamics of the Yangtze River

    NASA Astrophysics Data System (ADS)

    Cai, Huayang; Piccolroaz, Sebastiano; Huang, Jingzheng; Liu, Zhiyong; Liu, Feng; Toffolon, Marco

    2018-05-01

    This study examines the impact of the world’s largest dam, the Three Gorges Dam (TGD), on the thermal dynamics of the Yangtze River (China). The analysis uses long-term observations of river water temperature (RWT) in four stations and reconstructs the RWT that would have occurred in absence of the TGD. Relative to pre-TGD conditions, RWT consistently warmed in the region due to air temperature (AT) increase. In addition, the analysis demonstrates that the TGD significantly affected RWT in the downstream reach. At the closest downstream station (Yichang) to the TGD, the annual cycle of RWT experienced a damped response to AT and a marked seasonal alteration: warming during all seasons except for spring and early summer which were characterized by cooling. Both effects were a direct consequence of the larger thermal inertia of the massive water volume stored in the TGD reservoir, causing the downstream reach to be more thermally resilient. The approach used here to quantify the separate contributions of climate and human interventions on RWT can be used to set scientific guidelines for river management and conservation planning strategies.

  18. Evaluation of Gridded Precipitation Data for Driving SWAT Model in Area Upstream of Three Gorges Reservoir

    PubMed Central

    Yang, Yan; Wang, Guoqiang; Wang, Lijing; Yu, Jingshan; Xu, Zongxue

    2014-01-01

    Gridded precipitation data are becoming an important source for driving hydrologic models to achieve stable and valid simulation results in different regions. Thus, evaluating different sources of precipitation data is important for improving the applicability of gridded data. In this study, we used three gridded rainfall datasets: 1) National Centers for Environmental Prediction - Climate Forecast System Reanalysis (NCEP-CFSR); 2) Asian Precipitation - Highly-Resolved Observational Data Integration Towards Evaluation (APHRODITE); and 3) China trend - surface reanalysis (trend surface) data. These are compared with monitoring precipitation data for driving the Soil and Water Assessment Tool in two basins upstream of Three Gorges Reservoir (TGR) in China. The results of one test basin with significant topographic influence indicates that all the gridded data have poor abilities in reproducing hydrologic processes with the topographic influence on precipitation quantity and distribution. However, in a relatively flat test basin, the APHRODITE and trend surface data can give stable and desirable results. The results of this study suggest that precipitation data for future applications should be considered comprehensively in the TGR area, including the influence of data density and topography. PMID:25409467

  19. A DFT Study of Tungsten-Methylidene Formation on a W/ZSM-5 Zeolite: The Metathesis Active Site.

    PubMed

    Maihom, Thana; Probst, Michael; Limtrakul, Jumras

    2015-10-26

    Tungsten-methylidene formation from ethene on either the W(IV) , W(V) , or W(VI) active sites of a W/ZSM-5 zeolite is investigated by using the M06-L functional. The reaction is assumed to proceed in two steps; the first step is the [2+2] cycloaddition between ethene and the W-O active site to form an oxametallacycle intermediate. The intermediate is then decomposed to produce the W-methylidene active site from the metathesis reaction. The overall activation barrier of the reaction on W(VI) (27.3 kcal mol(-1) ) is considerably lower than the ones for W(IV) and W(V) (69.4 and 37.1 kcal mol(-1) , respectively). Moreover, the reaction involving the W(VI) site also stabilizes intermediates and products to a larger extent than the ones on the W(IV) and W(V) sites. As a result, we have demonstrated that the reaction of the W-methylidene metathesis active site is both kinetically and thermodynamically favored to occur on the W(VI) active site of the zeolite. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Artificial Metalloproteins Containing Co 4O 4Cubane Active Sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Olshansky, Lisa; Huerta-Lavorie, Raul; Nguyen, Andy I.

    Artificial metalloproteins (ArMs) containing Co 4O 4 cubane active sites were constructed via biotin-streptavidin technology. Stabilized by hydrogen bonds (H-bonds), terminal and cofacial Co III-OH 2 moieties are observed crystallographically in a series of immobilized cubane sites. Solution electrochemistry provided correlations of oxidation potential and pH. For variants containing Ser and Phe adjacent to the metallocofactor, 1e -/1H + chemistry predominates until pH 8, above which the oxidation becomes pH-independent. Installation of Tyr proximal to the Co 4O 4 active site provided a single H-bond to one of a set of cofacial Co III-OH 2 groups. With this variant, multi-emore » - /multi-H + chemistry is observed, along with a change in mechanism at pH 9.5 that is consistent with Tyr deprotonation. Finally, with structural similarities to both the oxygen-evolving complex of photosystem II (H-bonded Tyr) and to thin film water oxidation catalysts (Co 4O 4 core), these findings bridge synthetic and biological systems for water oxidation, highlighting the importance of secondary sphere interactions in mediating multi-e - /multi-H + reactivity.« less

  1. Artificial Metalloproteins Containing Co 4O 4Cubane Active Sites

    DOE PAGES

    Olshansky, Lisa; Huerta-Lavorie, Raul; Nguyen, Andy I.; ...

    2018-02-05

    Artificial metalloproteins (ArMs) containing Co 4O 4 cubane active sites were constructed via biotin-streptavidin technology. Stabilized by hydrogen bonds (H-bonds), terminal and cofacial Co III-OH 2 moieties are observed crystallographically in a series of immobilized cubane sites. Solution electrochemistry provided correlations of oxidation potential and pH. For variants containing Ser and Phe adjacent to the metallocofactor, 1e -/1H + chemistry predominates until pH 8, above which the oxidation becomes pH-independent. Installation of Tyr proximal to the Co 4O 4 active site provided a single H-bond to one of a set of cofacial Co III-OH 2 groups. With this variant, multi-emore » - /multi-H + chemistry is observed, along with a change in mechanism at pH 9.5 that is consistent with Tyr deprotonation. Finally, with structural similarities to both the oxygen-evolving complex of photosystem II (H-bonded Tyr) and to thin film water oxidation catalysts (Co 4O 4 core), these findings bridge synthetic and biological systems for water oxidation, highlighting the importance of secondary sphere interactions in mediating multi-e - /multi-H + reactivity.« less

  2. A population genetic analysis of the midget faded rattlesnake in Wyoming

    USGS Publications Warehouse

    Oyler-McCance, Sara J.; Parker, J.M.

    2010-01-01

    Little is known about the population biology of midget faded rattlesnakes, a sensitive subspecies of the Western Rattlesnake, despite conservation efforts to protect them. We conducted a molecular genetic study of midget faded rattlesnakes in southwestern Wyoming to investigate population genetic structure in this area, particularly with reference to Flaming Gorge Reservoir and its associated human activities, and to document levels of genetic diversity. We genotyped 229 snakes from 11 sampling sites using 9 microsatellite loci. We found significant levels of genetic structure among sites that were better explained by geographic region and isolation by distance than by position relative to waterways. Sites on either side of the reservoir at its widest point were not significantly different. Six of the sites showed signatures of a population bottleneck using an alpha value of 0.05. Three of these bottlenecked sites (the three most northern) were the most genetically distinct and occur in areas of greatest impact from human activity.

  3. The stakeholder preference for best management practices in the Three Gorges Reservoir Region.

    PubMed

    Qiu, Jiali; Shen, Zhenyao; Chen, Lei; Xie, Hui; Sun, Chengchun; Huang, Qin

    2014-11-01

    A qualitative analysis of in-depth interviews with 92 farmers and 42 policy managers in Wuxi County, the Three Gorges Reservoir Region, was conducted to identify stakeholder preferences for alternative best management practices (BMPs) and to determine the factors that affect their acceptance. Policy manager support for most of the practices was relatively stronger than support by farmers, except for the grade stabilization structure (GSS), hillside ditch (HD) and constructed wetland alternative, owing to their perceptions of soil benefits, economic advantages and environmental advantages. Farmers opposed those practices that occupied cultivated lands or changed the conventional planting methods, such as field border, conservation tillage (CT) and contour buffer strips. They tended to accept the BMPs with off-farm pollution reduction, such as GSS, riparian forest buffer and HD, and the BMPs associated with soil benefits, such as nutrient management and Terrace. The result that almost all respondents did not accept CT differed from reports in the existing literature. There is a significant correlation between the acceptance of some BMPs and the townships where the farmers lived (P ≤ 0.05). The environmental conditions and social factors would affect farmer support for BMPs, including local soil conditions, farming methods, economic income, education level and age. The economic advantages of the BMPs were the main motivation for farmers to accept the practices. Furthermore, intensive education efforts, financial incentives or economic subsidies may promote the adoption of the BMPs in our study area.

  4. The Stakeholder Preference for Best Management Practices in the Three Gorges Reservoir Region

    NASA Astrophysics Data System (ADS)

    Qiu, Jiali; Shen, Zhenyao; Chen, Lei; Xie, Hui; Sun, Chengchun; Huang, Qin

    2014-11-01

    A qualitative analysis of in-depth interviews with 92 farmers and 42 policy managers in Wuxi County, the Three Gorges Reservoir Region, was conducted to identify stakeholder preferences for alternative best management practices (BMPs) and to determine the factors that affect their acceptance. Policy manager support for most of the practices was relatively stronger than support by farmers, except for the grade stabilization structure (GSS), hillside ditch (HD) and constructed wetland alternative, owing to their perceptions of soil benefits, economic advantages and environmental advantages. Farmers opposed those practices that occupied cultivated lands or changed the conventional planting methods, such as field border, conservation tillage (CT) and contour buffer strips. They tended to accept the BMPs with off-farm pollution reduction, such as GSS, riparian forest buffer and HD, and the BMPs associated with soil benefits, such as nutrient management and Terrace. The result that almost all respondents did not accept CT differed from reports in the existing literature. There is a significant correlation between the acceptance of some BMPs and the townships where the farmers lived ( P ≤ 0.05). The environmental conditions and social factors would affect farmer support for BMPs, including local soil conditions, farming methods, economic income, education level and age. The economic advantages of the BMPs were the main motivation for farmers to accept the practices. Furthermore, intensive education efforts, financial incentives or economic subsidies may promote the adoption of the BMPs in our study area.

  5. New cholinesterase inhibitors from Garcinia atroviridis.

    PubMed

    Tan, Wen-Nee; Khairuddean, Melati; Wong, Keng-Chong; Khaw, Kooi-Yeong; Vikneswaran, Murugaiyah

    2014-09-01

    A triflavanone, Garcineflavanone A (1) and a biflavonol, Garcineflavonol A (2) have been isolated from the stem bark of Garcinia atroviridis (Clusiaceae), collected in Peninsular Malaysia. Their structures were established using one and two-dimensional NMR, UV, IR and mass spectrometry and evaluated in vitro for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes inhibitory activity. Molecular docking studies of the isolated compounds were performed using docking procedure of AutoDock to disclose the binding interaction and orientation of these molecules into the active site gorge. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Understanding Which Residues of the Active Site and Loop Structure of a Tyrosine Aminomutase Define Its Mutase and Lyase Activities.

    PubMed

    Attanayake, Gayanthi; Walter, Tyler; Walker, Kevin D

    2018-05-30

    Site-directed mutations and substrate analogues were used to gain insights into the branch-point reaction of the 3,5-dihydro-5-methylidene-4 H-imidazol-4-one (MIO)-tyrosine aminomutase from Oryza sativa ( OsTAM). Exchanging the active residues of OsTAM (Y125C/N446K) for those in a phenylalanine aminomutase TcPAM altered its substrate specificity from tyrosine to phenylalanine. The aminomutase mechanism of OsTAM surprisingly changed almost exclusively to that of an ammonia lyase making cinnamic acid (>95%) over β-phenylalanine [Walter, T., et al. (2016) Biochemistry 55, 3497-3503]. We hypothesized that the missing electronics or sterics on the aryl ring of the phenylalanine substrate, compared with the sizable electron-donating hydroxyl of the natural tyrosine substrate, influenced the unexpected lyase reactivity of the OsTAM mutant. The double mutant was incubated with 16 α-phenylalanine substituent analogues of varying electronic strengths and sterics. The mutant converted each analogue principally to its acrylate with ∼50% conversion of the p-Br substrate, making only a small amount of the β-amino acid. The inner loop structure over the entrance to the active site was also mutated to assess how the lyase and mutase activities are affected. An OsTAM loop mutant, matching the loop residues of TcPAM, still chiefly made >95% of the acrylate from each substrate. A combined active site:loop mutant was most reactive but remained a lyase, making 10-fold more acrylates than other mutants did. While mutations within the active site changed the substrate specificity of OsTAM, continued exploration is needed to fully understand the interplay among the inner loop, the substrate, and the active site in defining the mutase and lyase activities.

  7. Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    O'Dell, William B.; Agarwal, Pratul K.; Meilleur, Flora

    Lytic polysaccharide monooxygenases have attracted vast attention owing to their abilities to disrupt glycosidic bonds via oxidation instead of hydrolysis and to enhance enzymatic digestion of recalcitrant substrates including chitin and cellulose. Here, we determined the high-resolution X-ray crystal structures of an enzyme from Neurospora crassa in the resting state and of a copper(II) dioxo intermediate complex formed in the absence of substrate. X-ray crystal structures also revealed “pre-bound” molecular oxygen adjacent to the active site. An examination of protonation states enabled by neutron crystallography and density functional theory calculations identified a role for a conserved histidine in promoting oxygenmore » activation. Our results provide a new structural description of oxygen activation by substrate free lytic polysaccharide monooxygenases and provide insights that can be extended to reactivity in the enzyme–substrate complex.« less

  8. Oxygen Activation at the Active Site of a Fungal Lytic Polysaccharide Monooxygenase

    DOE PAGES

    O'Dell, William B.; Agarwal, Pratul K.; Meilleur, Flora

    2016-12-22

    Lytic polysaccharide monooxygenases have attracted vast attention owing to their abilities to disrupt glycosidic bonds via oxidation instead of hydrolysis and to enhance enzymatic digestion of recalcitrant substrates including chitin and cellulose. Here, we determined the high-resolution X-ray crystal structures of an enzyme from Neurospora crassa in the resting state and of a copper(II) dioxo intermediate complex formed in the absence of substrate. X-ray crystal structures also revealed “pre-bound” molecular oxygen adjacent to the active site. An examination of protonation states enabled by neutron crystallography and density functional theory calculations identified a role for a conserved histidine in promoting oxygenmore » activation. Our results provide a new structural description of oxygen activation by substrate free lytic polysaccharide monooxygenases and provide insights that can be extended to reactivity in the enzyme–substrate complex.« less

  9. Fragment-based identification of determinants of conformational and spectroscopic change at the ricin active site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Carra,J.; McHugh, C.; Mulligan, S.

    2007-01-01

    We found that amide ligands can bind weakly but specifically to the ricin active site, producing significant shifts in positions of the critical active site residues Arg180 and Tyr80. These results indicate that fragment-based drug discovery methods are capable of identifying minimal bonding determinants of active-site side-chain rearrangements and the mechanistic origins of spectroscopic shifts. Our results suggest that tryptophan fluorescence provides a sensitive probe for the geometric relationship of arginine-tryptophan pairs, which often have significant roles in protein function. Using the unusual characteristics of the RTA system, we measured the still controversial thermodynamic changes of site-specific urea binding tomore » a protein, results that are relevant to understanding the physical mechanisms of protein denaturation.« less

  10. Active Site Desolvation and Thermostability Trade-Offs in the Evolution of Catalytically Diverse Triazine Hydrolases.

    PubMed

    Sugrue, Elena; Carr, Paul D; Scott, Colin; Jackson, Colin J

    2016-11-15

    The desolvation of ionizable residues in the active sites of enzymes and the subsequent effects on catalysis and thermostability have been studied in model systems, yet little about how enzymes can naturally evolve to include active sites with highly reactive and desolvated charges is known. Variants of triazine hydrolase (TrzN) with significant differences in their active sites have been isolated from different bacterial strains: TrzN from Nocardioides sp. strain MTD22 contains a catalytic glutamate residue (Glu241) that is surrounded by hydrophobic and aromatic second-shell residues (Pro214 and Tyr215), whereas TrzN from Nocardioides sp. strain AN3 has a noncatalytic glutamine residue (Gln241) at an equivalent position, surrounded by hydrophilic residues (Thr214 and His215). To understand how and why these variants have evolved, a series of TrzN mutants were generated and characterized. These results show that desolvation by second-shell residues increases the pK a of Glu241, allowing it to act as a general acid at neutral pH. However, significant thermostability trade-offs are required to incorporate the ionizable Glu241 in the active site and to then enclose it in a hydrophobic microenvironment. Analysis of high-resolution crystal structures shows that there are almost no structural changes to the overall configuration of the active site due to these mutations, suggesting that the changes in activity and thermostability are purely based on the altered electrostatics. The natural evolution of these enzyme isoforms provides a unique system in which to study the fundamental process of charged residue desolvation in enzyme catalysis and its relative contribution to the creation and evolution of an enzyme active site.

  11. 76 FR 30696 - Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-26

    ... DEPARTMENT OF ENERGY Reimbursement for Costs of Remedial Action at Active Uranium and Thorium...) acceptance of claims in FY 2011 from eligible active uranium and thorium processing site licensees for... incurred by licensees at active uranium and thorium processing sites to remediate byproduct material...

  12. Active Site and Remote Contributions to Catalysis in Methylthioadenosine Nucleosidases

    DOE PAGES

    Thomas, Keisha; Cameron, Scott A.; Almo, Steven C.; ...

    2015-03-25

    5'-Methylthioadenosine/S-adenosyl-l-homocysteine nucleosidases (MTANs) catalyze the hydrolysis of 5'-methylthioadenosine to adenine and 5-methylthioribose. The amino acid sequences of the MTANs from Vibrio cholerae (VcMTAN) and Escherichia coli (EcMTAN) are 60% identical and 75% similar. Protein structure folds and kinetic properties are similar. However, binding of transition-state analogues is dominated by favorable entropy in VcMTAN and by enthalpy in EcMTAN. Catalytic sites of VcMTAN and EcMTAN in contact with reactants differ by two residues; Ala113 and Val153 in VcMTAN are Pro113 and Ile152, respectively, in EcMTAN. Here, we mutated the VcMTAN catalytic site residues to match those of EcMTAN in anticipation ofmore » altering its properties toward EcMTAN. Inhibition of VcMTAN by transition-state analogues required filling both active sites of the homodimer. However, in the Val153Ile mutant or double mutants, transition-state analogue binding at one site caused complete inhibition. Therefore, a single amino acid, Val153, alters the catalytic site cooperativity in VcMTAN. The transition-state analogue affinity and thermodynamics in mutant VcMTAN became even more unlike those of EcMTAN, the opposite of expectations from catalytic site similarity; thus, catalytic site contacts in VcMTAN are unable to recapitulate the properties of EcMTAN. X-ray crystal structures of EcMTAN, VcMTAN, and a multiple-site mutant of VcMTAN most closely resembling EcMTAN in catalytic site contacts show no major protein conformational differences. In conclusion, the overall protein architectures of these closely related proteins are implicated in contributing to the catalytic site differences.« less

  13. Active Site and Remote Contributions to Catalysis in Methylthioadenosine Nucleosidases

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Thomas, Keisha; Cameron, Scott A.; Almo, Steven C.

    5'-Methylthioadenosine/S-adenosyl-l-homocysteine nucleosidases (MTANs) catalyze the hydrolysis of 5'-methylthioadenosine to adenine and 5-methylthioribose. The amino acid sequences of the MTANs from Vibrio cholerae (VcMTAN) and Escherichia coli (EcMTAN) are 60% identical and 75% similar. Protein structure folds and kinetic properties are similar. However, binding of transition-state analogues is dominated by favorable entropy in VcMTAN and by enthalpy in EcMTAN. Catalytic sites of VcMTAN and EcMTAN in contact with reactants differ by two residues; Ala113 and Val153 in VcMTAN are Pro113 and Ile152, respectively, in EcMTAN. Here, we mutated the VcMTAN catalytic site residues to match those of EcMTAN in anticipation ofmore » altering its properties toward EcMTAN. Inhibition of VcMTAN by transition-state analogues required filling both active sites of the homodimer. However, in the Val153Ile mutant or double mutants, transition-state analogue binding at one site caused complete inhibition. Therefore, a single amino acid, Val153, alters the catalytic site cooperativity in VcMTAN. The transition-state analogue affinity and thermodynamics in mutant VcMTAN became even more unlike those of EcMTAN, the opposite of expectations from catalytic site similarity; thus, catalytic site contacts in VcMTAN are unable to recapitulate the properties of EcMTAN. X-ray crystal structures of EcMTAN, VcMTAN, and a multiple-site mutant of VcMTAN most closely resembling EcMTAN in catalytic site contacts show no major protein conformational differences. In conclusion, the overall protein architectures of these closely related proteins are implicated in contributing to the catalytic site differences.« less

  14. Disentangling Early Stone Age palimpsests: determining the functional independence of hominid- and carnivore-derived portions of archaeofaunas.

    PubMed

    Egeland, Charles P; Pickering, Travis Rayne; Domínguez-Rodrigo, Manuel; Brain, C K

    2004-11-01

    Determining the extent to which hominid- and carnivore-derived components of fossil bone palimpsests formed independently of each other can provide valuable information to paleoanthropologists interested in reconstructing the foraging adaptations of hominids. Because stone tool cutmarks, hammerstone percussion marks, and carnivore tooth marks are usually only imparted on bone during nutrient extraction from a carcass, these bone surface modifications are particularly amenable to the types of analyses that might meet this goal. This study compares the percentage of limb bone specimens that preserve evidence of both hominid- and carnivore-imparted bone damage from actualistic control samples and several Plio-Pleistocene archaeofaunas, including new data from Swartkrans Member 3 (South Africa). We argue that this procedure, which elucidates the degree of hominid-carnivore independence in assemblage formation, will allow researchers to extract for focused analyses high integrity components (hominid and carnivore) from presumably low integrity sites. Comparisons suggest that the hominid- and carnivore-derived components from sites in Olduvai Gorge Bed II (Tanzania), the ST Site Complex at Peninj (Tanzania), and Swartkrans Member 3 formed largely independent of each other, while data from the FLK 22 Zinjanthropus (FLK Zinj) site (Olduvai Gorge Bed I) indicate significant interdependence in assemblage formation. This contrast suggests that some Early Stone Age assemblages (e.g., the Olduvai Gorge Bed II sites, the Peninj ST Site Complex, and Swartkrans Member 3) are probably more useful than others (e.g., FLK Zinj) for assessing the maximal carcass-acquiring abilities of early hominids; in such assemblages as those in the former set, sole hominid-contribution is more confidently discerned and isolated for analysis than in assemblages such as FLK Zinj.

  15. Western Pond Turtle Head-starting and Reintroduction; 2003-2004 Annual Report.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Van Leuven, Susan; Allen, Harriet; Slavin, Kate

    This report covers the results of the western pond turtle head-starting and reintroduction project for the period of October 2003-September 2004. Wild hatchling western pond turtles from the Columbia River Gorge were reared at the Woodland Park and Oregon Zoos in 2003 and 2004 as part of the recovery effort for this Washington State endangered species. The objective of the program is to reduce losses to introduced predators like bullfrogs and largemouth bass by raising the hatchlings to a size where they are too large to be eaten by most of these predators. Sixty-nine turtles were over-wintered at the Woodlandmore » Park Zoo and 69 at the Oregon Zoo. Of these, 136 head-started juvenile turtles were released at three sites in the Columbia Gorge in 2004. Two were held back to attain more growth in captivity. Thirty-four were released at the Klickitat ponds, 19 at the Klickitat lake, 21 at the Skamania site, and 62 at Pierce National Wildlife Refuge (NWR). This brought the total number of head-start turtles released since 1991 to 246 for the Klickitat ponds, 114 for the Klickitat lake, 167 for the Skamania pond complex, and 250 at Pierce NWR. In 2004, 32 females from the two Columbia Gorge populations were equipped with transmitters and monitored for nesting activity. Twenty-one of the females nested and produced 85 hatchlings. The hatchlings were collected in September and October and transported to the Woodland Park and Oregon zoos for rearing in the head-start program. Data collection for a four-year telemetry study of survival and habitat use by juvenile western pond turtles at Pierce NWR concluded in 2004. Radio transmitters on study animals were replaced as needed until all replacements were in service; afterward, the turtles were monitored until their transmitters failed. The corps of study turtles ranged from 39 in August 2003 to 2 turtles at the end of August 2004. These turtles showed the same seasonal pattern of movements between summer water and upland

  16. Jack bean urease: the effect of active-site binding inhibitors on the reactivity of enzyme thiol groups.

    PubMed

    Krajewska, Barbara; Zaborska, Wiesława

    2007-10-01

    In view of the complexity of the role of the active site flap cysteine in the urease catalysis, in this work we studied how the presence of typical active-site binding inhibitors of urease, phenylphosphorodiamidate (PPD), acetohydroxamic acid (AHA), boric acid and fluoride, affects the reactivity of enzyme thiol groups, the active site flap thiol in particular. For that the inhibitor-urease complexes were prepared with excess inhibitors and had their thiol groups titrated with DTNB. The effects observed were analyzed in terms of the structures of the inhibitor-urease complexes reported in the literature. We found that the effectiveness in preventing the active site cysteine from the modification by disulfides, varied among the inhibitors studied, even though they all bind to the active site. The variations were accounted for by different extents of geometrical distortion in the active site that the inhibitors introduced upon binding, leaving the flap either open in AHA-, boric acid- and fluoride-inhibited urease, like in the native enzyme or closed in PPD-inhibited urease. Among the inhibitors, only PPD was found to be able to thoroughly protect the flap cysteines from the further reaction with disulfides, this apparently resulting from the closed conformation of the flap. Accordingly, in practical terms PPD may be regarded as the most suitable inhibitor for active-site protection experiments in inhibition studies of urease.

  17. Spatial and temporal assessment of driving and conditioning factors and their impact on land use / land cover change in the Xiangxi Catchment, Three Gorges Region

    NASA Astrophysics Data System (ADS)

    Seeber, Christoph; Hartmann, Heike; Xiang, Wei; King, Lorenz

    2010-05-01

    Land use / land cover change (LUCC) is the most important human alteration of the earth's surface and is primarily studied in cases where it leads to severe environmental problems. The construction of the Three Gorges Dam on the Yangtze River in China has an extensive impact on the ecosystems and the local population. To assess its impact, the Xiangxi Catchment is taken as an example. The outlet of the Xiangxi River, a northern tributary of the Yangtze River, is located about 40 km upstream of the Three Gorges Dam. Due to the loss of fertile arable land and residential land which is mainly induced by the inundation and measures of resettlement, enormous LUCC is observed in the study area by depicting the land use / land cover by classification of LandsatTM data retrieved in 1987 and 2007. LUCC in the Xiangxi Catchment during this period can generally be characterized as decrease of cultivated land, increase of woodland and fallow land, and a shift in cropping from traditional smallholder farming to the establishment of citrus orchards, which are implemented as cash crops. Not only the inundation and the resettlement have an impact on LUCC, also the newly built and improved traffic infrastructure, growth of urban structures and land use policies in terms of environmental protection are expected to play an important role concerning LUCC. To assess the spatial and temporal impact of influencing factors, a LUCC gradient is generated based on post-classification change analysis of multispectral data. Furthermore, inter-stages between 1987 and 2007 have to be examined, to reach for a higher temporal resolution, which shall help to figure out temporal relationships between LUCC and the occurrence of driving factors. Once influence factors and and their spatial and temporal impacts are identified, a basis for predicting LUCC in the future for is provided for this area.

  18. Stringency of the 2-His–1-Asp Active-Site Motif in Prolyl 4-Hydroxylase

    PubMed Central

    Gorres, Kelly L.; Pua, Khian Hong; Raines, Ronald T.

    2009-01-01

    The non-heme iron(II) dioxygenase family of enzymes contain a common 2-His–1-carboxylate iron-binding motif. These enzymes catalyze a wide variety of oxidative reactions, such as the hydroxylation of aliphatic C–H bonds. Prolyl 4-hydroxylase (P4H) is an α-ketoglutarate-dependent iron(II) dioxygenase that catalyzes the post-translational hydroxylation of proline residues in protocollagen strands, stabilizing the ensuing triple helix. Human P4H residues His412, Asp414, and His483 have been identified as an iron-coordinating 2-His–1-carboxylate motif. Enzymes that catalyze oxidative halogenation do so by a mechanism similar to that of P4H. These halogenases retain the active-site histidine residues, but the carboxylate ligand is replaced with a halide ion. We replaced Asp414 of P4H with alanine (to mimic the active site of a halogenase) and with glycine. These substitutions do not, however, convert P4H into a halogenase. Moreover, the hydroxylase activity of D414A P4H cannot be rescued with small molecules. In addition, rearranging the two His and one Asp residues in the active site eliminates hydroxylase activity. Our results demonstrate a high stringency for the iron-binding residues in the P4H active site. We conclude that P4H, which catalyzes an especially demanding chemical transformation, is recalcitrant to change. PMID:19890397

  19. Improving the neutral phytase activity from Bacillus amyloliquefaciens DSM 1061 by site-directed mutagenesis.

    PubMed

    Xu, Wei; Shao, Rong; Wang, Zupeng; Yan, Xiuhua

    2015-03-01

    Neutral phytase is used as a feed additive for degradation of anti-nutritional phytate in aquatic feed industry. Site-directed mutagenesis of Bacillus amyloliquefaciens DSM 1061 phytase was performed with an aim to increase its activity. Mutation residues were chosen based on multiple sequence alignments and structure analysis of neutral phytsaes from different microorganisms. The mutation sites on surface (D148E, S197E and N156E) and around the active site (D52E) of phytase were selected. Analysis of the phytase variants showed that the specific activities of mutants D148E and S197E remarkably increased by about 35 and 13% over a temperature range of 40-75 °C at pH 7.0, respectively. The k cat of mutants D148E and S197E were 1.50 and 1.25 times than that of the wild-type phytase, respectively. Both D148E and S197E showed much higher thermostability than that of the wild-type phytase. However, mutants N156E and D52E led to significant loss of specific activity of the enzyme. Structural analysis revealed that these mutations may affect conformation of the active site of phytase. The present mutant phytases D148E and S197E with increased activities and thermostabilities have application potential as additives in aquaculture feed.

  20. Transfer hydrogenation over sodium-modified ceria: Enrichment of redox sites active for alcohol dehydrogenation

    DOE PAGES

    Nelson, Nicholas C.; Boote, Brett W.; Naik, Pranjali; ...

    2017-01-17

    Ceria (CeO 2) and sodium-modified ceria (Ce-Na) were prepared through combustion synthesis. Palladium was deposited onto the supports (Pd/CeO 2 and Pd/Ce-Na) and their activity for the aqueous-phase transfer hydrogenation of phenol using 2-propanol under liquid flow conditions was studied. Pd/Ce-Na showed a marked increase (6×) in transfer hydrogenation activity over Pd/CeO 2. Material characterization indicated that water-stable sodium species were not doped into the ceria lattice, but rather existed as subsurface carbonates. Modification of ceria by sodium provided more adsorption and redox active sites (i.e. defects) for 2-propanol dehydrogenation. This effect was an intrinsic property of the Ce-Na supportmore » and independent of Pd. The redox sites active for 2-propanol dehydrogenation were thermodynamically equivalent on both supports/catalysts. At high phenol concentrations, the reaction was limited by 2-propanol adsorption. Furthermore, the difference in catalytic activity was attributed to the different numbers of 2-propanol adsorption and redox active sites on each catalyst.« less

  1. Improved ethanol electrooxidation performance by shortening Pd-Ni active site distance in Pd-Ni-P nanocatalysts

    NASA Astrophysics Data System (ADS)

    Chen, Lin; Lu, Lilin; Zhu, Hengli; Chen, Yueguang; Huang, Yu; Li, Yadong; Wang, Leyu

    2017-01-01

    Incorporating oxophilic metals into noble metal-based catalysts represents an emerging strategy to improve the catalytic performance of electrocatalysts in fuel cells. However, effects of the distance between the noble metal and oxophilic metal active sites on the catalytic performance have rarely been investigated. Herein, we report on ultrasmall (~5 nm) Pd-Ni-P ternary nanoparticles for ethanol electrooxidation. The activity is improved up to 4.95 A per mgPd, which is 6.88 times higher than commercial Pd/C (0.72 A per mgPd), by shortening the distance between Pd and Ni active sites, achieved through shape transformation from Pd/Ni-P heterodimers into Pd-Ni-P nanoparticles and tuning the Ni/Pd atomic ratio to 1:1. Density functional theory calculations reveal that the improved activity and stability stems from the promoted production of free OH radicals (on Ni active sites) which facilitate the oxidative removal of carbonaceous poison and combination with CH3CO radicals on adjacent Pd active sites.

  2. Testing the applicability of rapid on-site enzymatic activity detection for surface water monitoring

    NASA Astrophysics Data System (ADS)

    Stadler, Philipp; Vogl, Wolfgang; Juri, Koschelnik; Markus, Epp; Maximilian, Lackner; Markus, Oismüller; Monika, Kumpan; Peter, Strauss; Regina, Sommer; Gabriela, Ryzinska-Paier; Farnleitner Andreas, H.; Matthias, Zessner

    2015-04-01

    On-site detection of enzymatic activities has been suggested as a rapid surrogate for microbiological pollution monitoring of water resources (e.g. using glucuronidases, galactosidases, esterases). Due to the possible short measuring intervals enzymatic methods have high potential as near-real time water quality monitoring tools. This presentation describes results from a long termed field test. For twelve months, two ColiMinder devices (Vienna Water Monitoring, Austria) for on-site determination of enzymatic activity were tested for stream water monitoring at the experimental catchment HOAL (Hydrological Open Air Laboratory, Center for Water Resource Systems, Vienna University of Technology). The devices were overall able to follow and reflect the diverse hydrological and microbiological conditions of the monitored stream during the test period. Continuous data in high temporal resolution captured the course of enzymatic activity in stream water during diverse rainfall events. The method also proofed sensitive enough to determine diurnal fluctuations of enzymatic activity in stream water during dry periods. The method was able to capture a seasonal trend of enzymatic activity in stream water that matches the results gained from Colilert18 analysis for E. coli and coliform bacteria of monthly grab samples. Furthermore the comparison of ColiMinder data with measurements gained at the same test site with devices using the same method but having different construction design (BACTcontrol, microLAN) showed consistent measuring results. Comparative analysis showed significant differences between measured enzymatic activity (modified fishman units and pmol/min/100ml) and cultivation based analyses (most probable number, colony forming unit). Methods of enzymatic activity measures are capable to detect ideally the enzymatic activity caused by all active target bacteria members, including VBNC (viable but nonculturable) while cultivation based methods cannot detect VBNC

  3. Agonist activation of α7 nicotinic acetylcholine receptors via an allosteric transmembrane site

    PubMed Central

    Gill, JasKiran K.; Savolainen, Mari; Young, Gareth T.; Zwart, Ruud; Sher, Emanuele; Millar, Neil S.

    2011-01-01

    Conventional nicotinic acetylcholine receptor (nAChR) agonists, such as acetylcholine, act at an extracellular “orthosteric” binding site located at the interface between two adjacent subunits. Here, we present evidence of potent activation of α7 nAChRs via an allosteric transmembrane site. Previous studies have identified a series of nAChR-positive allosteric modulators (PAMs) that lack agonist activity but are able to potentiate responses to orthosteric agonists, such as acetylcholine. It has been shown, for example, that TQS acts as a conventional α7 nAChR PAM. In contrast, we have found that a compound with close chemical similarity to TQS (4BP-TQS) is a potent allosteric agonist of α7 nAChRs. Whereas the α7 nAChR antagonist metyllycaconitine acts competitively with conventional nicotinic agonists, metyllycaconitine is a noncompetitive antagonist of 4BP-TQS. Mutation of an amino acid (M253L), located in a transmembrane cavity that has been proposed as being the binding site for PAMs, completely blocks agonist activation by 4BP-TQS. In contrast, this mutation had no significant effect on agonist activation by acetylcholine. Conversely, mutation of an amino acid located within the known orthosteric binding site (W148F) has a profound effect on agonist potency of acetylcholine (resulting in a shift of ∼200-fold in the acetylcholine dose-response curve), but had little effect on the agonist dose-response curve for 4BP-TQS. Computer docking studies with an α7 homology model provides evidence that both TQS and 4BP-TQS bind within an intrasubunit transmembrane cavity. Taken together, these findings provide evidence that agonist activation of nAChRs can occur via an allosteric transmembrane site. PMID:21436053

  4. A remote palm domain residue of RB69 DNA polymerase is critical for enzyme activity and influences the conformation of the active site.

    PubMed

    Jacewicz, Agata; Trzemecka, Anna; Guja, Kip E; Plochocka, Danuta; Yakubovskaya, Elena; Bebenek, Anna; Garcia-Diaz, Miguel

    2013-01-01

    Non-conserved amino acids that are far removed from the active site can sometimes have an unexpected effect on enzyme catalysis. We have investigated the effects of alanine replacement of residues distant from the active site of the replicative RB69 DNA polymerase, and identified a substitution in a weakly conserved palm residue (D714A), that renders the enzyme incapable of sustaining phage replication in vivo. D714, located several angstroms away from the active site, does not contact the DNA or the incoming dNTP, and our apoenzyme and ternary crystal structures of the Pol(D714A) mutant demonstrate that D714A does not affect the overall structure of the protein. The structures reveal a conformational change of several amino acid side chains, which cascade out from the site of the substitution towards the catalytic center, substantially perturbing the geometry of the active site. Consistent with these structural observations, the mutant has a significantly reduced k pol for correct incorporation. We propose that the observed structural changes underlie the severe polymerization defect and thus D714 is a remote, non-catalytic residue that is nevertheless critical for maintaining an optimal active site conformation. This represents a striking example of an action-at-a-distance interaction.

  5. Responses of landscape pattern of China's two largest freshwater lakes to early dry season after the impoundment of Three-Gorges Dam

    NASA Astrophysics Data System (ADS)

    Wu, Haipeng; Zeng, Guangming; Liang, Jie; Chen, Jin; Xu, Jijun; Dai, Juan; Sang, Lianhai; Li, Xiaodong; Ye, Shujing

    2017-04-01

    The effects of hydrologic cycle change (caused by human activity and global climate change) on ecosystems attract the increasing attention around the world. As a result of impounding of the Three Gorges Dam (TGD), climate change and sand mining, the dry season of Poyang Lake and Dongting Lake (China's two largest freshwater lakes) came early after the TGD impoundment. It was the primary cause of the increasing need for sluice/dam construction to store water in the Lakes and attracted increasing attention. In this paper, we compared the landscape pattern between three hydrologic years with early dry season (EY) and three normal hydrologic years (NY) of each lake by remote sensing technology, to reveal the effect of early dry season on landscape pattern. The results showed that early dry season caused expanding of Phalaris to mudflat zone in Poyang Lake, while caused expanding of Carex to Phalaris zone and expanding of Phalaris to mudflat zone in Dongting Lake. In landscape level, there was no significant difference in landscape grain size, landscape grain shape, habitat connectivity and landscape diversity between EY and NY in the two lakes. While in habitat class level, there were significant changes in area of mudflat and Phalaris and grain size of mudflat in Poyang Lake, and in area of Carex, grain size of Phalaris and grain shape of Carex and Phalaris in Dongting Lake. These changes will impact migrating birds of East Asian and migratory fishes of Yangtze River.

  6. Work-site wellness programmes in Sweden: a cross-sectional study of physical activity, self-efficacy, and health.

    PubMed

    Gånedahl, H; Zsaludek Viklund, P; Carlén, K; Kylberg, E; Ekberg, J

    2015-05-01

    In Sweden, a work-site wellness programme implies reimbursing some of the expenses for health-promoting activities. Although work-site wellness programmes are readily available in Sweden, a large number of employees elect not to participate. The aim of this study was to investigate the association of physical activity, self-reported general health assessment and self-efficacy with participation in a work-site wellness programme. A cross-sectional study design was used. An online questionnaire was distributed to employees of a manufacturing company with 2500 employees in southwest Sweden. Those who took advantage of the work-site wellness programme assessed their general health as better and had higher assessment of physical activity. The study showed that being enlisted also implies a higher level of physical activity and general health; however, the effect sizes of these correlations were small. Self-efficacy, i.e. perceived behavioural control, was not associated with participation in the work-site wellness programme. However, self-efficacy was correlated with both general health assessment and physical activity. A regression analysis to determine explanatory contributions to the general health assessment score showed no significant contribution from participation in a work-site wellness programme, but was instead explained by perceived behavioural control and physical activity. Given the small effect size of the difference in physical activity between participators and non-participators in the work-site wellness programme, it is probable that only a small proportion of participators changed their health-promoting activities as a result of the work-site wellness programme. Copyright © 2015 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.

  7. Activity of N-coordinated multi-metal-atom active site structures for Pt-free oxygen reduction reaction catalysis: Role of *OH ligands

    DOE PAGES

    Holby, Edward F.; Taylor, Christopher D.

    2015-03-19

    We report calculated oxygen reduction reaction energy pathways on multi-metal-atom structures that have previously been shown to be thermodynamically favorable. We predict that such sites have the ability to spontaneously cleave the O₂ bond and then will proceed to over-bind reaction intermediates. In particular, the *OH bound state has lower energy than the final 2 H₂O state at positive potentials. Contrary to traditional surface catalysts, this *OH binding does not poison the multi-metal-atom site but acts as a modifying ligand that will spontaneously form in aqueous environments leading to new active sites that have higher catalytic activities. These *OH boundmore » structures have the highest calculated activity to date.« less

  8. The Diverse AAA+ Machines that Repair Inhibited Rubisco Active Sites

    PubMed Central

    Mueller-Cajar, Oliver

    2017-01-01

    Gaseous carbon dioxide enters the biosphere almost exclusively via the active site of the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco). This highly conserved catalyst has an almost universal propensity to non-productively interact with its substrate ribulose 1,5-bisphosphate, leading to the formation of dead-end inhibited complexes. In diverse autotrophic organisms this tendency has been counteracted by the recruitment of dedicated AAA+ (ATPases associated with various cellular activities) proteins that all use the energy of ATP hydrolysis to remodel inhibited Rubisco active sites leading to release of the inhibitor. Three evolutionarily distinct classes of these Rubisco activases (Rcas) have been discovered so far. Green and red-type Rca are mostly found in photosynthetic eukaryotes of the green and red plastid lineage respectively, whereas CbbQO is associated with chemoautotrophic bacteria. Ongoing mechanistic studies are elucidating how the various motors are utilizing both similar and contrasting strategies to ultimately perform their common function of cracking the inhibited Rubisco active site. The best studied mechanism utilized by red-type Rca appears to involve transient threading of the Rubisco large subunit C-terminal peptide, reminiscent of the action performed by Clp proteases. As well as providing a fascinating example of convergent molecular evolution, Rca proteins can be considered promising crop-improvement targets. Approaches aiming to replace Rubisco in plants with improved enzymes will need to ensure the presence of a compatible Rca protein. The thermolability of the Rca protein found in crop plants provides an opportunity to fortify photosynthesis against high temperature stress. Photosynthesis also appears to be limited by Rca when light conditions are fluctuating. Synthetic biology strategies aiming to enhance the autotrophic CO2 fixation machinery will need to take into consideration the requirement for Rubisco activases

  9. Diversity and above-ground biomass patterns of vascular flora induced by flooding in the drawdown area of China's Three Gorges Reservoir.

    PubMed

    Wang, Qiang; Yuan, Xingzhong; Willison, J H Martin; Zhang, Yuewei; Liu, Hong

    2014-01-01

    Hydrological alternation can dramatically influence riparian environments and shape riparian vegetation zonation. However, it was difficult to predict the status in the drawdown area of the Three Gorges Reservoir (TGR), because the hydrological regime created by the dam involves both short periods of summer flooding and long-term winter impoundment for half a year. In order to examine the effects of hydrological alternation on plant diversity and biomass in the drawdown area of TGR, twelve sites distributed along the length of the drawdown area of TGR were chosen to explore the lateral pattern of plant diversity and above-ground biomass at the ends of growing seasons in 2009 and 2010. We recorded 175 vascular plant species in 2009 and 127 in 2010, indicating that a significant loss of vascular flora in the drawdown area of TGR resulted from the new hydrological regimes. Cynodon dactylon and Cyperus rotundus had high tolerance to short periods of summer flooding and long-term winter flooding. Almost half of the remnant species were annuals. Species richness, Shannon-Wiener Index and above-ground biomass of vegetation exhibited an increasing pattern along the elevation gradient, being greater at higher elevations subjected to lower submergence stress. Plant diversity, above-ground biomass and species distribution were significantly influenced by the duration of submergence relative to elevation in both summer and previous winter. Several million tonnes of vegetation would be accumulated on the drawdown area of TGR in every summer and some adverse environmental problems may be introduced when it was submerged in winter. We conclude that vascular flora biodiversity in the drawdown area of TGR has dramatically declined after the impoundment to full capacity. The new hydrological condition, characterized by long-term winter flooding and short periods of summer flooding, determined vegetation biodiversity and above-ground biomass patterns along the elevation gradient in

  10. Diversity and Above-Ground Biomass Patterns of Vascular Flora Induced by Flooding in the Drawdown Area of China's Three Gorges Reservoir

    PubMed Central

    Wang, Qiang; Yuan, Xingzhong; Willison, J.H.Martin; Zhang, Yuewei; Liu, Hong

    2014-01-01

    Hydrological alternation can dramatically influence riparian environments and shape riparian vegetation zonation. However, it was difficult to predict the status in the drawdown area of the Three Gorges Reservoir (TGR), because the hydrological regime created by the dam involves both short periods of summer flooding and long-term winter impoundment for half a year. In order to examine the effects of hydrological alternation on plant diversity and biomass in the drawdown area of TGR, twelve sites distributed along the length of the drawdown area of TGR were chosen to explore the lateral pattern of plant diversity and above-ground biomass at the ends of growing seasons in 2009 and 2010. We recorded 175 vascular plant species in 2009 and 127 in 2010, indicating that a significant loss of vascular flora in the drawdown area of TGR resulted from the new hydrological regimes. Cynodon dactylon and Cyperus rotundus had high tolerance to short periods of summer flooding and long-term winter flooding. Almost half of the remnant species were annuals. Species richness, Shannon-Wiener Index and above-ground biomass of vegetation exhibited an increasing pattern along the elevation gradient, being greater at higher elevations subjected to lower submergence stress. Plant diversity, above-ground biomass and species distribution were significantly influenced by the duration of submergence relative to elevation in both summer and previous winter. Several million tonnes of vegetation would be accumulated on the drawdown area of TGR in every summer and some adverse environmental problems may be introduced when it was submerged in winter. We conclude that vascular flora biodiversity in the drawdown area of TGR has dramatically declined after the impoundment to full capacity. The new hydrological condition, characterized by long-term winter flooding and short periods of summer flooding, determined vegetation biodiversity and above-ground biomass patterns along the elevation gradient in

  11. Differential regulation of the transcriptional activity of the glucocorticoid receptor through site-specific phosphorylation.

    PubMed

    Kumar, Raj; Calhoun, William J

    2008-12-01

    Post-translational modifications such as phosphorylation are known to play an important role in the gene regulation by the transcription factors including the nuclear hormone receptor superfamily of which the glucocorticoid receptor (GR) is a member. Protein phosphorylation often switches cellular activity from one state to another. Like many other transcription factors, the GR is a phosphoprotein, and phosphorylation plays an important role in the regulation of GR activity. Cell signaling pathways that regulate phosphorylation of the GR and its associated proteins are important determinants of GR function under various physiological conditions. While the role of many phosphorylation sites in the GR is still not fully understood, the role of others is clearer. Several aspects of transcription factor function, including DNA binding affinity, interaction of transactivation domains with the transcription initiation complex, and shuttling between the cytoplasmic compartments, have all been linked to site-specific phosphorylation. All major phosphorylation sites in the human GR are located in the N-terminal domain including the major transactivation domain, AF1. Available literature clearly indicates that many of these potential phosphorylation sites are substrates for multiple kinases, suggesting the potential for a very complex regulatory network. Phosphorylated GR interacts favorably with critical coregulatory proteins and subsequently enhances transcriptional activity. In addition, the activities and specificities of coregulators may be subject to similar regulation by phosphorylation. Regulation of the GR activity due to phosphorylation appears to be site-specific and dependent upon specific cell signaling cascade. Taken together, site-specific phosphorylation and related kinase pathways play an important role in the action of the GR, and more precise mechanistic information will lead to fuller understanding of the complex nature of gene regulation by the GR- and

  12. MX Siting Investigation. Water Resources Program Industry Activity Inventory, Nevada-Utah.

    DTIC Science & Technology

    1980-09-02

    sites. New and revived mining activities and the cooling needs of possible new coal -fired electric power plants represent the chief competitors with MX...34 !- ---- ON CO. Figure .-. Ma showing araipce yUaIoto f𔃻XMsieCmlx 1 3 include new mining activity and coal -fired, geothermal, and hydroelectric j energy...in northeastern Juab County. The Soil Conservation Service has been actively pushing land treatment programs to increase the productivity of irrigated

  13. Enthalpic Breakdown of Water Structure on Protein Active-Site Surfaces

    PubMed Central

    Haider, Kamran; Wickstrom, Lauren; Ramsey, Steven; Gilson, Michael K.; Kurtzman, Tom

    2016-01-01

    The principles underlying water reorganization around simple non-polar solutes are well understood and provide the framework for classical hydrophobic effect, whereby water molecules structure themselves around solutes so that they maintain favorable energetic contacts with both the solute and with other water molecules. However, for certain solute surface topographies, water molecules, due to their geometry and size, are unable to simultaneously maintain favorable energetic contacts with both the surface and neighboring water molecules. In this study, we analyze the solvation of ligand-binding sites for six structurally diverse proteins using hydration site analysis and measures of local water structure, in order to identify surfaces at which water molecules are unable to structure themselves in a way that maintains favorable enthalpy relative to bulk water. These surfaces are characterized by a high degree of enclosure, weak solute-water interactions, and surface constraints that induce unfavorable pair interactions between neighboring water molecules. Additionally, we find that the solvation of charged side-chains in an active site generally results in favorable enthalpy but can also lead to pair interactions between neighboring water molecules that are significantly unfavorable relative to bulk water. We find that frustrated local structure can occur not only in apolar and weakly polar pockets, where overall enthalpy tends to be unfavorable, but also in charged pockets, where overall water enthalpy tends to be favorable. The characterization of local water structure in these terms may prove useful for evaluating the displacement of water from diverse protein active-site environments. PMID:27169482

  14. Nevada National Security Site Industrial Sites Project Closeout - 12498

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cabble, Kevin; Krauss, Mark; Matthews, Pat

    The U.S. Department of Energy (DOE), National Nuclear Security Administration Nevada Site Office is responsible for environmental restoration (ER) at the Nevada National Security Site (NNSS). This includes remediation at Industrial Sites where past nuclear testing activities and activities that supported nuclear testing may have or are known to have resulted in the release of contaminants into the environment. Industrial Sites at the NNSS have included nuclear facilities that supported the nuclear rocket/missile development programs, gas stations, landfills, spill sites, ordnance sites, and numerous other waste disposal and release sites. The NNSS Industrial Sites activities neared completion at the endmore » of fiscal year 2011 while other activities required under the Federal Facility Agreement and Consent Order (FFACO) and part of the same NNSS ER Project are forecasted to extend to 2027 or beyond. With the majority of Industrial Sites corrective action units (CAUs) completed (more than 250 CAUs and over 1,800 corrective action sites), it was determined that an activity closeout process should be implemented to ensure that the work completed over the past 15 years is well documented in a comprehensive and concise summary. While the process used to close each individual CAU is described in approved documents, no single document describes in summary fashion the work completed to close the many individual Industrial Sites. The activity closeout process will be used to develop an Industrial Sites closeout document that describes these years of work. This document will summarize the number of Industrial Sites closed under the FFACO and provide general descriptions of projects, contaminants removed, and sites closed in place with corresponding Use Restrictions. Other pertinent information related to Industrial Sites work such as the project history, closure decisions, historical declarations, remediation strategies, and final CAU status will be included in the

  15. Dual Active Site in the Endolytic Transglycosylase gp144 of Bacteriophage phiKZ.

    PubMed

    Chertkov, O V; Armeev, G A; Uporov, I V; Legotsky, S A; Sykilinda, N N; Shaytan, A K; Klyachko, N L; Miroshnikov, K A

    2017-01-01

    Lytic transglycosylases are abundant peptidoglycan lysing enzymes that degrade the heteropolymers of bacterial cell walls in metabolic processes or in the course of a bacteriophage infection. The conventional catalytic mechanism of transglycosylases involves only the Glu or Asp residue. Endolysin gp144 of Pseudomonas aeruginosa bacteriophage phiKZ belongs to the family of Gram-negative transglycosylases with a modular composition and C -terminal location of the catalytic domain. Glu115 of gp144 performs the predicted role of a catalytic residue. However, replacement of this residue does not completely eliminate the activity of the mutant protein. Site-directed mutagenesis has revealed the participation of Tyr197 in the catalytic mechanism, as well as the presence of a second active site involving Glu178 and Tyr147. The existence of the dual active site was supported by computer modeling and monitoring of the molecular dynamics of the changes in the conformation and surface charge distribution as a consequence of point mutations.

  16. Active sites for CO 2 hydrogenation to methanol on Cu/ZnO catalysts

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kattel, Shyam; Ramírez, Pedro J.; Chen, Jingguang G.

    The active sites over commercial copper/zinc oxide/aluminum oxide (Cu/ZnO/Al 2O 3) catalysts for carbon dioxide (CO 2) hydrogenation to methanol, the Zn-Cu bimetallic sites or ZnO-Cu interfacial sites, have recently been the subject of intense debate. Here, we report a direct comparison between the activity of ZnCu and ZnO/Cu model catalysts for methanol synthesis. By combining x-ray photoemission spectroscopy, density functional theory, and kinetic Monte Carlo simulations, we can identify and characterize the reactivity of each catalyst. Both experimental and theoretical results agree that ZnCu undergoes surface oxidation under the reaction conditions so that surface Zn transforms into ZnO andmore » allows ZnCu to reach the activity of ZnO/Cu with the same Zn coverage. These results highlight a synergy of Cu and ZnO at the interface that facilitates methanol synthesis via formate intermediates.« less

  17. Active sites for CO 2 hydrogenation to methanol on Cu/ZnO catalysts

    DOE PAGES

    Kattel, Shyam; Ramírez, Pedro J.; Chen, Jingguang G.; ...

    2017-03-23

    The active sites over commercial copper/zinc oxide/aluminum oxide (Cu/ZnO/Al 2O 3) catalysts for carbon dioxide (CO 2) hydrogenation to methanol, the Zn-Cu bimetallic sites or ZnO-Cu interfacial sites, have recently been the subject of intense debate. Here, we report a direct comparison between the activity of ZnCu and ZnO/Cu model catalysts for methanol synthesis. By combining x-ray photoemission spectroscopy, density functional theory, and kinetic Monte Carlo simulations, we can identify and characterize the reactivity of each catalyst. Both experimental and theoretical results agree that ZnCu undergoes surface oxidation under the reaction conditions so that surface Zn transforms into ZnO andmore » allows ZnCu to reach the activity of ZnO/Cu with the same Zn coverage. These results highlight a synergy of Cu and ZnO at the interface that facilitates methanol synthesis via formate intermediates.« less

  18. Influence of a Confined Methanol Solvent on the Reactivity of Active Sites in UiO-66.

    PubMed

    Caratelli, Chiara; Hajek, Julianna; Rogge, Sven M J; Vandenbrande, Steven; Meijer, Evert Jan; Waroquier, Michel; Van Speybroeck, Veronique

    2018-02-19

    UiO-66, composed of Zr-oxide bricks and terephthalate linkers, is currently one of the most studied metal-organic frameworks due to its exceptional stability. Defects can be introduced in the structure, creating undercoordinated Zr atoms which are Lewis acid sites. Here, additional Brønsted sites can be generated by coordinated protic species from the solvent. In this Article, a multilevel modeling approach was applied to unravel the effect of a confined methanol solvent on the active sites in UiO-66. First, active sites were explored with static periodic density functional theory calculations to investigate adsorption of water and methanol. Solvent was then introduced in the pores with grand canonical Monte Carlo simulations, followed by a series of molecular dynamics simulations at operating conditions. A hydrogen-bonded network of methanol molecules is formed, allowing the protons to shuttle between solvent methanol, adsorbed water, and the inorganic brick. Upon deprotonation of an active site, the methanol solvent aids the transfer of protons and stabilizes charged configurations via hydrogen bonding, which could be crucial in stabilizing reactive intermediates. The multilevel modeling approach adopted here sheds light on the important role of a confined solvent on the active sites in the UiO-66 material, introducing dynamic acidity in the system at finite temperatures by which protons may be easily shuttled from various positions at the active sites. © 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  19. Impact of single-site axonal GABAergic synaptic events on cerebellar interneuron activity.

    PubMed

    de San Martin, Javier Zorrilla; Jalil, Abdelali; Trigo, Federico F

    2015-12-01

    Axonal ionotropic receptors are present in a variety of neuronal types, and their function has largely been associated with the modulation of axonal activity and synaptic release. It is usually assumed that activation of axonal GABA(A)Rs comes from spillover, but in cerebellar molecular layer interneurons (MLIs) the GABA source is different: in these cells, GABA release activates presynaptic GABA(A) autoreceptors (autoRs) together with postsynaptic targets, producing an autoR-mediated synaptic event. The frequency of presynaptic, autoR-mediated miniature currents is twice that of their somatodendritic counterparts, suggesting that autoR-mediated responses have an important effect on interneuron activity. Here, we used local Ca(2+) photolysis in MLI axons of juvenile rats to evoke GABA release from individual varicosities to study the activation of axonal autoRs in single release sites. Our data show that single-site autoR conductances are similar to postsynaptic dendritic conductances. In conditions of high [Cl(-)](i), autoR-mediated conductances range from 1 to 5 nS; this corresponds to ∼30-150 GABA(A) channels per presynaptic varicosity, a value close to the number of channels in postsynaptic densities. Voltage responses produced by the activation of autoRs in single varicosities are amplified by a Na(v)-dependent mechanism and propagate along the axon with a length constant of 91 µm. Immunolabeling determination of synapse location shows that on average, one third of the synapses produce autoR-mediated signals that are large enough to reach the axon initial segment. Finally, we show that single-site activation of presynaptic GABA(A) autoRs leads to an increase in MLI excitability and thus conveys a strong feedback signal that contributes to spiking activity. © 2015 Zorrilla de San Martin et al.

  20. Impact of single-site axonal GABAergic synaptic events on cerebellar interneuron activity

    PubMed Central

    Zorrilla de San Martin, Javier; Jalil, Abdelali

    2015-01-01

    Axonal ionotropic receptors are present in a variety of neuronal types, and their function has largely been associated with the modulation of axonal activity and synaptic release. It is usually assumed that activation of axonal GABAARs comes from spillover, but in cerebellar molecular layer interneurons (MLIs) the GABA source is different: in these cells, GABA release activates presynaptic GABAA autoreceptors (autoRs) together with postsynaptic targets, producing an autoR-mediated synaptic event. The frequency of presynaptic, autoR-mediated miniature currents is twice that of their somatodendritic counterparts, suggesting that autoR-mediated responses have an important effect on interneuron activity. Here, we used local Ca2+ photolysis in MLI axons of juvenile rats to evoke GABA release from individual varicosities to study the activation of axonal autoRs in single release sites. Our data show that single-site autoR conductances are similar to postsynaptic dendritic conductances. In conditions of high [Cl−]i, autoR-mediated conductances range from 1 to 5 nS; this corresponds to ∼30–150 GABAA channels per presynaptic varicosity, a value close to the number of channels in postsynaptic densities. Voltage responses produced by the activation of autoRs in single varicosities are amplified by a Nav-dependent mechanism and propagate along the axon with a length constant of 91 µm. Immunolabeling determination of synapse location shows that on average, one third of the synapses produce autoR-mediated signals that are large enough to reach the axon initial segment. Finally, we show that single-site activation of presynaptic GABAA autoRs leads to an increase in MLI excitability and thus conveys a strong feedback signal that contributes to spiking activity. PMID:26621773

  1. Recent Experience Using Active Love Wave Techniques to Characterize Seismographic Station Sites

    NASA Astrophysics Data System (ADS)

    Martin, A. J.; Yong, A.; Salomone, L.

    2014-12-01

    Active-source Love waves recorded by the multi-channel analysis of surface wave (MASLW) technique were recently analyzed in two site characterization projects. Between 2010 and 2011, the 2009 American Recovery and Reinvestment Act (ARRA) funded GEOVision to conduct geophysical investigations at 189 seismographic stations—185 in California and 4 in the Central Eastern U.S. (CEUS). The original project plan was to utilize active and passive Rayleigh wave-based techniques to obtain shear-wave velocity (VS) profiles to a minimum depth of 30 m and the time-averaged VS of the upper 30 meters (VS30). Early in the investigation it became evident that Rayleigh wave techniques, such as multi-channel analysis of surface waves (MASRW), were not effective at characterizing all sites. Shear-wave seismic refraction and MASLW techniques were therefore applied. The MASLW technique was deployed at a total of 38 sites, in addition to other methods, and used as the primary technique to characterize 22 sites, 5 of which were also characterized using Rayleigh wave techniques. In 2012, the Electric Power Research Institute funded characterization of 33 CEUS station sites. Based on experience from the ARRA investigation, both MASRW and MASLW data were acquired by GEOVision at 24 CEUS sites—the remaining 9 sites and 2 overlapping sites were characterized by University of Texas, Austin. Of the 24 sites characterized by GEOVision, 16 were characterized using MASLW data, 4 using both MASLW and MASRW data and 4 using MASRW data. Love wave techniques were often found to perform better, or at least yield phase velocity data that could be more readily modeled using the fundamental mode assumption, at shallow rock sites, sites with steep velocity gradients, and, sites with a thin, low velocity, surficial soil layer overlying stiffer sediments. These types of velocity structure often excite dominant higher modes in Rayleigh wave data, but not in Love wave data. At such sites, it may be possible

  2. Human 15-LOX-1 active site mutations alter inhibitor binding and decrease potency.

    PubMed

    Armstrong, Michelle; van Hoorebeke, Christopher; Horn, Thomas; Deschamps, Joshua; Freedman, J Cody; Kalyanaraman, Chakrapani; Jacobson, Matthew P; Holman, Theodore

    2016-11-01

    Human 15-lipoxygenase-1 (h15-LOX-1 or h12/15-LOX) reacts with polyunsaturated fatty acids and produces bioactive lipid derivatives that are implicated in many important human diseases. One such disease is stroke, which is the fifth leading cause of death and the first leading cause of disability in America. The discovery of h15-LOX-1 inhibitors could potentially lead to novel therapeutics in the treatment of stroke, however, little is known about the inhibitor/active site interaction. This study utilizes site-directed mutagenesis, guided in part by molecular modeling, to gain a better structural understanding of inhibitor interactions within the active site. We have generated eight mutants (R402L, R404L, F414I, F414W, E356Q, Q547L, L407A, I417A) of h15-LOX-1 to determine whether these active site residues interact with two h15-LOX-1 inhibitors, ML351 and an ML094 derivative, compound 18. IC 50 values and steady-state inhibition kinetics were determined for the eight mutants, with four of the mutants affecting inhibitor potency relative to wild type h15-LOX-1 (F414I, F414W, E356Q and L407A). The data indicate that ML351 and compound 18, bind in a similar manner in the active site to an aromatic pocket close to F414 but have subtle differences in their specific binding modes. This information establishes the binding mode for ML094 and ML351 and will be leveraged to develop next-generation inhibitors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Active Site Detection by Spatial Conformity and Electrostatic Analysis—Unravelling a Proteolytic Function in Shrimp Alkaline Phosphatase

    PubMed Central

    Chakraborty, Sandeep; Minda, Renu; Salaye, Lipika; Bhattacharjee, Swapan K.; Rao, Basuthkar J.

    2011-01-01

    Computational methods are increasingly gaining importance as an aid in identifying active sites. Mostly these methods tend to have structural information that supplement sequence conservation based analyses. Development of tools that compute electrostatic potentials has further improved our ability to better characterize the active site residues in proteins. We have described a computational methodology for detecting active sites based on structural and electrostatic conformity - C ata L ytic A ctive S ite P rediction (CLASP). In our pipelined model, physical 3D signature of any particular enzymatic function as defined by its active sites is used to obtain spatially congruent matches. While previous work has revealed that catalytic residues have large pKa deviations from standard values, we show that for a given enzymatic activity, electrostatic potential difference (PD) between analogous residue pairs in an active site taken from different proteins of the same family are similar. False positives in spatially congruent matches are further pruned by PD analysis where cognate pairs with large deviations are rejected. We first present the results of active site prediction by CLASP for two enzymatic activities - β-lactamases and serine proteases, two of the most extensively investigated enzymes. The results of CLASP analysis on motifs extracted from Catalytic Site Atlas (CSA) are also presented in order to demonstrate its ability to accurately classify any protein, putative or otherwise, with known structure. The source code and database is made available at www.sanchak.com/clasp/. Subsequently, we probed alkaline phosphatases (AP), one of the well known promiscuous enzymes, for additional activities. Such a search has led us to predict a hitherto unknown function of shrimp alkaline phosphatase (SAP), where the protein acts as a protease. Finally, we present experimental evidence of the prediction by CLASP by showing that SAP indeed has protease activity in vitro

  4. The role of the peripheral benzodiazepine receptor in photodynamic activity of certain pyropheophorbide ether photosensitizers: albumin site II as a surrogate marker for activity.

    PubMed

    Dougherty, Thomas J; Sumlin, Adam B; Greco, William R; Weishaupt, Kenneth R; Vaughan, Lurine A; Pandey, Ravindra K

    2002-07-01

    A study has been carried out to define the importance of the peripheral benzodiazepine receptor (PBR) as a binding site for a series of chlorin-type photosensitizers, pyropheophorbide-a ethers, the subject of a previous quantitative structure-activity relationship study by us. The effects of the PBR ligand PK11195 on the photodynamic activity have been determined in vivo for certain members of this series of alkyl-substituted ethers: two of the most active derivatives (hexyl and heptyl), the least active derivative (dodecyl [C12]) and one of intermediate activity (octyl [C8]). The photodynamic therapy (PDT) effect was inhibited by PK11195 for both of the most active derivatives, but no effect on PDT activity was found for the less active C12 or C8 ethers. The inhibitory effects of PK11195 were predicted by the binding of only the active derivatives to the benzodiazepine site on albumin, ie. human serum albumin (HSA)-Site II. Thus, as with certain other types of photosensitizers, it has been demonstrated with this series of pyropheophorbide ethers that in vitro binding to HSA-Site II is a predictor of both optimal in vivo activity and binding to the PBR in vivo.

  5. Enzyme Active Site Interactions by Raman/FTIR, NMR, and Ab Initio Calculations

    PubMed Central

    Deng, Hua

    2017-01-01

    Characterization of enzyme active site structure and interactions at high resolution is important for the understanding of the enzyme catalysis. Vibrational frequency and NMR chemical shift measurements of enzyme-bound ligands are often used for such purpose when X-ray structures are not available or when higher resolution active site structures are desired. This review is focused on how ab initio calculations may be integrated with vibrational and NMR chemical shift measurements to quantitatively determine high-resolution ligand structures (up to 0.001 Å for bond length and 0.01 Å for hydrogen bonding distance) and how interaction energies between bound ligand and its surroundings at the active site may be determined. Quantitative characterization of substrate ionic states, bond polarizations, tautomeric forms, conformational changes and its interactions with surroundings in enzyme complexes that mimic ground state or transition state can provide snapshots for visualizing the substrate structural evolution along enzyme-catalyzed reaction pathway. Our results have shown that the integration of spectroscopic studies with theoretical computation greatly enhances our ability to interpret experimental data and significantly increases the reliability of the theoretical analysis. PMID:24018325

  6. Improved ethanol electrooxidation performance by shortening Pd–Ni active site distance in Pd–Ni–P nanocatalysts

    PubMed Central

    Chen, Lin; Lu, Lilin; Zhu, Hengli; Chen, Yueguang; Huang, Yu; Li, Yadong; Wang, Leyu

    2017-01-01

    Incorporating oxophilic metals into noble metal-based catalysts represents an emerging strategy to improve the catalytic performance of electrocatalysts in fuel cells. However, effects of the distance between the noble metal and oxophilic metal active sites on the catalytic performance have rarely been investigated. Herein, we report on ultrasmall (∼5 nm) Pd–Ni–P ternary nanoparticles for ethanol electrooxidation. The activity is improved up to 4.95 A per mgPd, which is 6.88 times higher than commercial Pd/C (0.72 A per mgPd), by shortening the distance between Pd and Ni active sites, achieved through shape transformation from Pd/Ni–P heterodimers into Pd–Ni–P nanoparticles and tuning the Ni/Pd atomic ratio to 1:1. Density functional theory calculations reveal that the improved activity and stability stems from the promoted production of free OH radicals (on Ni active sites) which facilitate the oxidative removal of carbonaceous poison and combination with CH3CO radicals on adjacent Pd active sites. PMID:28071650

  7. Dithiolato-bridged nickel-iron complexes as models for the active site of [NiFe]-hydrogenases.

    PubMed

    Song, Li-Cheng; Yang, Xi-Yue; Cao, Meng; Gao, Xiu-Yun; Liu, Bei-Bei; Zhu, Liang; Jiang, Feng

    2017-03-30

    The structural and functional modeling of the active site of [NiFe]-hydrogenases has been proved to be challenging to a great extent. Herein, we report the synthesis, structures, and some properties of the NiFe-based dicarbonyl, terminal hydride, and μ-hydroxo models for the active site of [NiFe]-hydrogenases.

  8. A Bayesian-based two-stage inexact optimization method for supporting stream water quality management in the Three Gorges Reservoir region.

    PubMed

    Hu, X H; Li, Y P; Huang, G H; Zhuang, X W; Ding, X W

    2016-05-01

    In this study, a Bayesian-based two-stage inexact optimization (BTIO) method is developed for supporting water quality management through coupling Bayesian analysis with interval two-stage stochastic programming (ITSP). The BTIO method is capable of addressing uncertainties caused by insufficient inputs in water quality model as well as uncertainties expressed as probabilistic distributions and interval numbers. The BTIO method is applied to a real case of water quality management for the Xiangxi River basin in the Three Gorges Reservoir region to seek optimal water quality management schemes under various uncertainties. Interval solutions for production patterns under a range of probabilistic water quality constraints have been generated. Results obtained demonstrate compromises between the system benefit and the system failure risk due to inherent uncertainties that exist in various system components. Moreover, information about pollutant emission is accomplished, which would help managers to adjust production patterns of regional industry and local policies considering interactions of water quality requirement, economic benefit, and industry structure.

  9. Use of an Improved Matching Algorithm to Select Scaffolds for Enzyme Design Based on a Complex Active Site Model.

    PubMed

    Huang, Xiaoqiang; Xue, Jing; Lin, Min; Zhu, Yushan

    2016-01-01

    Active site preorganization helps native enzymes electrostatically stabilize the transition state better than the ground state for their primary substrates and achieve significant rate enhancement. In this report, we hypothesize that a complex active site model for active site preorganization modeling should help to create preorganized active site design and afford higher starting activities towards target reactions. Our matching algorithm ProdaMatch was improved by invoking effective pruning strategies and the native active sites for ten scaffolds in a benchmark test set were reproduced. The root-mean squared deviations between the matched transition states and those in the crystal structures were < 1.0 Å for the ten scaffolds, and the repacking calculation results showed that 91% of the hydrogen bonds within the active sites are recovered, indicating that the active sites can be preorganized based on the predicted positions of transition states. The application of the complex active site model for de novo enzyme design was evaluated by scaffold selection using a classic catalytic triad motif for the hydrolysis of p-nitrophenyl acetate. Eighty scaffolds were identified from a scaffold library with 1,491 proteins and four scaffolds were native esterase. Furthermore, enzyme design for complicated substrates was investigated for the hydrolysis of cephalexin using scaffold selection based on two different catalytic motifs. Only three scaffolds were identified from the scaffold library by virtue of the classic catalytic triad-based motif. In contrast, 40 scaffolds were identified using a more flexible, but still preorganized catalytic motif, where one scaffold corresponded to the α-amino acid ester hydrolase that catalyzes the hydrolysis and synthesis of cephalexin. Thus, the complex active site modeling approach for de novo enzyme design with the aid of the improved ProdaMatch program is a promising approach for the creation of active sites with high catalytic

  10. Use of an Improved Matching Algorithm to Select Scaffolds for Enzyme Design Based on a Complex Active Site Model

    PubMed Central

    Huang, Xiaoqiang; Xue, Jing; Lin, Min; Zhu, Yushan

    2016-01-01

    Active site preorganization helps native enzymes electrostatically stabilize the transition state better than the ground state for their primary substrates and achieve significant rate enhancement. In this report, we hypothesize that a complex active site model for active site preorganization modeling should help to create preorganized active site design and afford higher starting activities towards target reactions. Our matching algorithm ProdaMatch was improved by invoking effective pruning strategies and the native active sites for ten scaffolds in a benchmark test set were reproduced. The root-mean squared deviations between the matched transition states and those in the crystal structures were < 1.0 Å for the ten scaffolds, and the repacking calculation results showed that 91% of the hydrogen bonds within the active sites are recovered, indicating that the active sites can be preorganized based on the predicted positions of transition states. The application of the complex active site model for de novo enzyme design was evaluated by scaffold selection using a classic catalytic triad motif for the hydrolysis of p-nitrophenyl acetate. Eighty scaffolds were identified from a scaffold library with 1,491 proteins and four scaffolds were native esterase. Furthermore, enzyme design for complicated substrates was investigated for the hydrolysis of cephalexin using scaffold selection based on two different catalytic motifs. Only three scaffolds were identified from the scaffold library by virtue of the classic catalytic triad-based motif. In contrast, 40 scaffolds were identified using a more flexible, but still preorganized catalytic motif, where one scaffold corresponded to the α-amino acid ester hydrolase that catalyzes the hydrolysis and synthesis of cephalexin. Thus, the complex active site modeling approach for de novo enzyme design with the aid of the improved ProdaMatch program is a promising approach for the creation of active sites with high catalytic

  11. Chemical Modification of Papain and Subtilisin: An Active Site Comparison

    ERIC Educational Resources Information Center

    St-Vincent, Mireille; Dickman, Michael

    2004-01-01

    An experiment using methyle methanethiosulfonate (MMTS) and phenylmethylsulfonyl flouride (PMSF) to specifically modify the cysteine and serine residues in the active sites of papain and subtilism respectively is demonstrated. The covalent modification of these enzymes and subsequent rescue of papain shows the beginning biochemist that proteins…

  12. Comparative evaluation of light-trap catches, electric motor mosquito catches and human biting catches of Anopheles in the Three Gorges Reservoir.

    PubMed

    Duo-quan, Wang; Lin-hua, Tang; Zhen-cheng, Gu; Xiang, Zheng; Man-ni, Yang; Wei-kang, Jiang

    2012-01-01

    The mosquito sampling efficiency of light-trap catches and electric motor mosquito catches were compared with that of human biting catches in the Three Gorges Reservoir. There was consistency in the sampling efficiency between light-trap catches and human biting catches for Anopheles sinensis (r = 0.82, P<0.01) and light-trap catches were 1.52 (1.35-1.71) times that of human biting catches regardless of mosquito density (r = 0.33, P>0.01), while the correlation between electric motor mosquito catches and human biting catches was found to be not statistically significant (r = 0.43, P>0.01) and its sampling efficiency was below that of human biting catches. It is concluded that light-traps can be used as an alternative to human biting catches of Anopheles sinensis in the study area and is a promising tool for sampling malaria vector populations.

  13. Active-site copper reduction promotes substrate binding of fungal lytic polysaccharide monooxygenase and reduces stability.

    PubMed

    Kracher, Daniel; Andlar, Martina; Furtmüller, Paul G; Ludwig, Roland

    2018-02-02

    Lytic polysaccharide monooxygenases (LPMOs) are a class of copper-containing enzymes that oxidatively degrade insoluble plant polysaccharides and soluble oligosaccharides. Upon reductive activation, they cleave the substrate and promote biomass degradation by hydrolytic enzymes. In this study, we employed LPMO9C from Neurospora crassa , which is active toward cellulose and soluble β-glucans, to study the enzyme-substrate interaction and thermal stability. Binding studies showed that the reduction of the mononuclear active-site copper by ascorbic acid increased the affinity and the maximum binding capacity of LPMO for cellulose. The reduced redox state of the active-site copper and not the subsequent formation of the activated oxygen species increased the affinity toward cellulose. The lower affinity of oxidized LPMO could support its desorption after catalysis and allow hydrolases to access the cleavage site. It also suggests that the copper reduction is not necessarily performed in the substrate-bound state of LPMO. Differential scanning fluorimetry showed a stabilizing effect of the substrates cellulose and xyloglucan on the apparent transition midpoint temperature of the reduced, catalytically active enzyme. Oxidative auto-inactivation and destabilization were observed in the absence of a suitable substrate. Our data reveal the determinants of LPMO stability under turnover and non-turnover conditions and indicate that the reduction of the active-site copper initiates substrate binding. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Critical Amino Acids in the Active Site of Meprin Metalloproteinases for Substrate and Peptide Bond Specificity*

    PubMed Central

    Villa, James P.; Bertenshaw, Greg P.; Bond, Judith S.

    2008-01-01

    SUMMARY The protease domains of the evolutionarily-related α and ß subunits of meprin metalloproteases are approximately 55% identical at the amino acid level, however, their substrate and peptide bond specificities differ markedly. The meprin ß subunit favors acidic residues proximal to the scissile bond, while the α subunit prefers small or aromatic amino acids flanking the scissile bond. Thus gastrin, a peptide that contains a string of five Glu residues, is an excellent substrate for meprin ß while it is not hydrolyzed by meprin α. Work herein aimed to identify critical amino acids in the meprin active sites that determine the substrate specificity differences. Sequence alignments and homology models, based on the crystal structure of the crayfish astacin, showed electrostatic differences within the meprin active sites. Site-directed mutagenesis of active site residues demonstrated that replacement of a hydrophobic residue by a basic amino acid enabled the meprin α protease to cleave gastrin. The meprin αY199K mutant was most effective; the corresponding mutation of meprin ßK185Y resulted in decreased activity toward gastrin. Peptide cleavage site determinations and kinetic analyses using a variety of peptides extended evidence that meprin αTyr199/ßLys185 are substrate specificity determinants in meprin active sites. These studies shed light on the molecular basis for the substrate specificity differences of astacin metalloproteinases. PMID:12888571

  15. Impact of socioeconomic and meteorological factors on reservoirs' air quality: a case in the Three Gorges Reservoir of Chongqing (TGRC), China over a 10-year period.

    PubMed

    Peng, Ying; Zhou, Fengwu; Cui, Jian; Du, Ke; Leng, Qiangmei; Yang, Fumo; Chan, Andy; Zhao, Hongting

    2017-07-01

    The Three Gorges Dam's construction and industrial transfer have resulted in a new air pollution pattern with the potential to threaten the reservoir eco-environment. To assess the impact of socioeconomic factors on the pattern of air quality vairation and economical risks, concentrations of SO 2 , NO 2 , and PM 10 , industry genres, and meteorological conditions were selected in the Three Gorges Reservoir of Chongqing (TGRC) during 2006-2015. Results showed that air quality had improved to some extent, but atmospheric NO 2 showed an increased trend during 2011-2015. Spatially, higher atmospheric NO 2 extended to the surrounding area. The primary industry, especially for agriculture, had shown to be responsible for the remarkable increase of atmospheric NO 2 (p < 0.01) due to the direct burning of agricultural straws and the emission of livestock breeding. The improvement of regional industrial structure and industrialization benefited air pollutant reductions, but construction industries had inhibited the improvement of regional air quality. In the tertiary industry, the cargo industry at ports had significantly decreased atmospheric NO 2 as a result of eliminating the obsoleted small ships. Contrarily, the highway transportation had brought more air pollutants. The relative humidity was shown to be the main meteorological factor, which had an extremely remarkable relation with atmospheric SO 2 (p < 0.01) and a significant correlation with atmospheric NO 2 (p < 0.05), respectively. In the future, the development of agriculture and livestock breeding would make regional air quality improvement difficult, and atmospheric SO 2 , NO 2 , and PM 10 deposition would aggravate regional soil and water acidification and reactivate heavy metal in soil and sediment, further to pose a high level of ecological risk in the TGRC and other countries with reservoirs in the world.

  16. How Force Might Activate Talin's Vinculin Binding Sites: SMD Reveals a Structural Mechanism

    PubMed Central

    Hytönen, Vesa P; Vogel, Viola

    2008-01-01

    Upon cell adhesion, talin physically couples the cytoskeleton via integrins to the extracellular matrix, and subsequent vinculin recruitment is enhanced by locally applied tensile force. Since the vinculin binding (VB) sites are buried in the talin rod under equilibrium conditions, the structural mechanism of how vinculin binding to talin is force-activated remains unknown. Taken together with experimental data, a biphasic vinculin binding model, as derived from steered molecular dynamics, provides high resolution structural insights how tensile mechanical force applied to the talin rod fragment (residues 486–889 constituting helices H1–H12) might activate the VB sites. Fragmentation of the rod into three helix subbundles is prerequisite to the sequential exposure of VB helices to water. Finally, unfolding of a VB helix into a completely stretched polypeptide might inhibit further binding of vinculin. The first events in fracturing the H1–H12 rods of talin1 and talin2 in subbundles are similar. The proposed force-activated α-helix swapping mechanism by which vinculin binding sites in talin rods are exposed works distinctly different from that of other force-activated bonds, including catch bonds. PMID:18282082

  17. Identification of the fatty acid activation site on human ClC-2.

    PubMed

    Cuppoletti, John; Tewari, Kirti P; Chakrabarti, Jayati; Malinowska, Danuta H

    2017-06-01

    Fatty acids (including lubiprostone and cobiprostone) are human ClC-2 (hClC-2) Cl - channel activators. Molecular and cellular mechanisms underlying this activation were examined. Role of a four-amino acid PKA activation site, RGET 691 , of hClC-2 was investigated using wild-type (WT) and mutant (AGET, RGEA, and AGAA) hClC-2 expressed in 293EBNA cells as well as involvement of PKA, intracellular cAMP concentration ([cAMP] i ), EP 2 , or EP 4 receptor agonist activity. All fatty acids [lubiprostone, cobiprostone, eicosatetraynoic acid (ETYA), oleic acid, and elaidic acid] caused significant rightward shifts in concentration-dependent Cl - current activation (increasing EC 50 s) with mutant compared with WT hClC-2 channels, without changing time and voltage dependence, current-voltage rectification, or methadone inhibition of the channel. As with lubiprostone, cobiprostone activation of hClC-2 occurred with PKA inhibitor (myristoylated protein kinase inhibitor) present or when using double PKA activation site (RRAA 655 /RGEA 691 ) mutant. Cobiprostone did not activate human CFTR. Fatty acids did not increase [cAMP] i in hClC-2/293EBNA or T84 cells. Using T84 CFTR knockdown cells, cobiprostone increased hClC-2 Cl - currents without increasing [cAMP] i, while PGE 2 and forskolin-IBMX increased both. Fatty acids were not agonists of EP 2 or EP 4 receptors. L-161,982, a supposed EP 4 -selective inhibitor, had no effect on lubiprostone-activated hClC-2 Cl - currents but significantly decreased T84 cell barrier function measured by transepithelial resistance and fluorescent dextran transepithelial movement. The present findings show that RGET 691 of hClC-2 (possible binding site) plays an important functional role in fatty acid activation of hClC-2. PKA, [cAMP] i , and EP 2 or EP 4 receptors are not involved. These studies provide the molecular basis for fatty acid regulation of hClC-2. Copyright © 2017 the American Physiological Society.

  18. Computational active site analysis of molecular pathways to improve functional classification of enzymes.

    PubMed

    Ozyurt, A Sinem; Selby, Thomas L

    2008-07-01

    This study describes a method to computationally assess the function of homologous enzymes through small molecule binding interaction energy. Three experimentally determined X-ray structures and four enzyme models from ornithine cyclo-deaminase, alanine dehydrogenase, and mu-crystallin were used in combination with nine small molecules to derive a function score (FS) for each enzyme-model combination. While energy values varied for a single molecule-enzyme combination due to differences in the active sites, we observe that the binding energies for the entire pathway were proportional for each set of small molecules investigated. This proportionality of energies for a reaction pathway appears to be dependent on the amino acids in the active site and their direct interactions with the small molecules, which allows a function score (FS) to be calculated to assess the specificity of each enzyme. Potential of mean force (PMF) calculations were used to obtain the energies, and the resulting FS values demonstrate that a measurement of function may be obtained using differences between these PMF values. Additionally, limitations of this method are discussed based on: (a) larger substrates with significant conformational flexibility; (b) low homology enzymes; and (c) open active sites. This method should be useful in accurately predicting specificity for single enzymes that have multiple steps in their reactions and in high throughput computational methods to accurately annotate uncharacterized proteins based on active site interaction analysis. 2008 Wiley-Liss, Inc.

  19. Elucidating Oxygen Reduction Active Sites in Pyrolyzed Metal–Nitrogen Coordinated Non-Precious-Metal Electrocatalyst Systems

    PubMed Central

    2015-01-01

    Detailed understanding of the nature of the active centers in non-precious-metal-based electrocatalyst, and their role in oxygen reduction reaction (ORR) mechanistic pathways will have a profound effect on successful commercialization of emission-free energy devices such as fuel cells. Recently, using pyrolyzed model structures of iron porphyrins, we have demonstrated that a covalent integration of the Fe–Nx sites into π-conjugated carbon basal plane modifies electron donating/withdrawing capability of the carbonaceous ligand, consequently improving ORR activity. Here, we employ a combination of in situ X-ray spectroscopy and electrochemical methods to identify the various structural and functional forms of the active centers in non-heme Fe/N/C catalysts. Both methods corroboratively confirm the single site 2e– × 2e– mechanism in alkaline media on the primary Fe2+–N4 centers and the dual-site 2e– × 2e– mechanism in acid media with the significant role of the surface bound coexisting Fe/FexOy nanoparticles (NPs) as the secondary active sites. PMID:24817921

  20. Elucidating Oxygen Reduction Active Sites in Pyrolyzed Metal-Nitrogen Coordinated Non-Precious-Metal Electrocatalyst Systems.

    PubMed

    Tylus, Urszula; Jia, Qingying; Strickland, Kara; Ramaswamy, Nagappan; Serov, Alexey; Atanassov, Plamen; Mukerjee, Sanjeev

    2014-05-01

    Detailed understanding of the nature of the active centers in non-precious-metal-based electrocatalyst, and their role in oxygen reduction reaction (ORR) mechanistic pathways will have a profound effect on successful commercialization of emission-free energy devices such as fuel cells. Recently, using pyrolyzed model structures of iron porphyrins, we have demonstrated that a covalent integration of the Fe-N x sites into π-conjugated carbon basal plane modifies electron donating/withdrawing capability of the carbonaceous ligand, consequently improving ORR activity. Here, we employ a combination of in situ X-ray spectroscopy and electrochemical methods to identify the various structural and functional forms of the active centers in non-heme Fe/N/C catalysts. Both methods corroboratively confirm the single site 2e - × 2e - mechanism in alkaline media on the primary Fe 2+ -N 4 centers and the dual-site 2e - × 2e - mechanism in acid media with the significant role of the surface bound coexisting Fe/Fe x O y nanoparticles (NPs) as the secondary active sites.

  1. Linde FUSRAP Site Remediation: Engineering Challenges and Solutions of Remedial Activities on an Active Industrial Facility - 13506

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Beres, Christopher M.; Fort, E. Joseph; Boyle, James D.

    2013-07-01

    The Linde FUSRAP Site (Linde) is located in Tonawanda, New York at a major research and development facility for Praxair, Inc. (Praxair). Successful remediation activities at Linde combines meeting cleanup objectives of radiological contamination while minimizing impacts to Praxair business operations. The unique use of Praxair's property coupled with an array of active and abandoned utilities poses many engineering and operational challenges; each of which has been overcome during the remedial action at Linde. The U.S. Army Corps of Engineers - Buffalo District (USACE) and CABRERA SERVICES, INC. (CABRERA) have successfully faced engineering challenges such as relocation of an abovegroundmore » structure, structural protection of an active water line, and installation of active mechanical, electrical, and communication utilities to perform remediation. As remediation nears completion, continued success of engineering challenges is critical as remaining activities exist in the vicinity of infrastructure essential to business operations; an electrical substation and duct bank providing power throughout the Praxair facility. Emphasis on engineering and operations through final remediation and into site restoration will allow for the safe and successful completion of the project. (authors)« less

  2. Greenhouse Gas Fluxes at the Tablelands, NL, Canada: A Site of Active Serpentinization

    NASA Astrophysics Data System (ADS)

    Morrill, P. L.; Morrissey, L. S.; Cumming, E.

    2016-12-01

    Active sites of serpentinization have been proposed as sites for carbon capture and storage (CCS) projects. However, in addition to their ability to convert carbon dioxide to carbonate rock, sites of serpentinization also have the potential release methane, which is a more power greenhouse gas than carbon dioxide. Very little is known about the natural flux of carbon dioxide sequestered and methane released into the atmosphere from active sites of serpentinization. In this study we measured carbon dioxide, methane, and nitrous oxide gas fluxes at a pool of ultra-basic water discharging from serpentinized rock in Winterhouse Canyon, Gros Morne, Newfoundland. We found that the flux of methane released was 4.6 x 10-7 mol/m2/min and the carbon dioxide sequestered was 1.9 x 10-5 mol/m2/min, while the concentrations of nitrous oxide showed little change. Based on these fluxes we calculated predictive climate change parameters such as net radiative forcing and global warming potential which predicted that despite the methane being released the site still had an overall long-term atmospheric cooling effect based on the natural rate of carbon dioxide sequestration.

  3. Quantum delocalization of protons in the hydrogen-bond network of an enzyme active site.

    PubMed

    Wang, Lu; Fried, Stephen D; Boxer, Steven G; Markland, Thomas E

    2014-12-30

    Enzymes use protein architectures to create highly specialized structural motifs that can greatly enhance the rates of complex chemical transformations. Here, we use experiments, combined with ab initio simulations that exactly include nuclear quantum effects, to show that a triad of strongly hydrogen-bonded tyrosine residues within the active site of the enzyme ketosteroid isomerase (KSI) facilitates quantum proton delocalization. This delocalization dramatically stabilizes the deprotonation of an active-site tyrosine residue, resulting in a very large isotope effect on its acidity. When an intermediate analog is docked, it is incorporated into the hydrogen-bond network, giving rise to extended quantum proton delocalization in the active site. These results shed light on the role of nuclear quantum effects in the hydrogen-bond network that stabilizes the reactive intermediate of KSI, and the behavior of protons in biological systems containing strong hydrogen bonds.

  4. Substituting Tyr138 in the active site loop of human phenylalanine hydroxylase affects catalysis and substrate activation.

    PubMed

    Leandro, João; Stokka, Anne J; Teigen, Knut; Andersen, Ole A; Flatmark, Torgeir

    2017-07-01

    Mammalian phenylalanine hydroxylase (PAH) is a key enzyme in l-phenylalanine (l-Phe) metabolism and is active as a homotetramer. Biochemical and biophysical work has demonstrated that it cycles between two states with a variably low and a high activity, and that the substrate l-Phe is the key player in this transition. X-ray structures of the catalytic domain have shown mobility of a partially intrinsically disordered Tyr 138 -loop to the active site in the presence of l-Phe. The mechanism by which the loop dynamics are coupled to substrate binding at the active site in tetrameric PAH is not fully understood. We have here conducted functional studies of four Tyr 138 point mutants. A high linear correlation ( r 2 = 0.99) was observed between their effects on the catalytic efficiency of the catalytic domain dimers and the corresponding effect on the catalytic efficiency of substrate-activated full-length tetramers. In the tetramers, a correlation ( r 2 = 0.96) was also observed between the increase in catalytic efficiency (activation) and the global conformational change (surface plasmon resonance signal response) at the same l-Phe concentration. The new data support a similar functional importance of the Tyr 138 -loop in the catalytic domain and the full-length enzyme homotetramer.

  5. Inhibitor-based validation of a homology model of the active-site of tripeptidyl peptidase II.

    PubMed

    De Winter, Hans; Breslin, Henry; Miskowski, Tamara; Kavash, Robert; Somers, Marijke

    2005-04-01

    A homology model of the active site region of tripeptidyl peptidase II (TPP II) was constructed based on the crystal structures of four subtilisin-like templates. The resulting model was subsequently validated by judging expectations of the model versus observed activities for a broad set of prepared TPP II inhibitors. The structure-activity relationships observed for the prepared TPP II inhibitors correlated nicely with the structural details of the TPP II active site model, supporting the validity of this model and its usefulness for structure-based drug design and pharmacophore searching experiments.

  6. Effects of mutations on active site conformation and dynamics of RNA-dependent RNA polymerase from Coxsackievirus B3.

    PubMed

    Shen, Hujun; Deng, Mingsen; Zhang, Yachao

    2017-10-01

    Recent crystal structures of RNA-dependent RNA polymerase (3D pol ) from Coxsackievirus B3 (CVB3) revealed that a tyrosine mutation at Phe364 (F364Y) resulted in structures with open active site whereas a hydrophobic mutation at Phe364 (F364A) led to conformations with closed active site. Besides, the crystal structures showed that the F364W mutation had no preference between the open and closed active sites, similar to wild-type. In this paper, we present a molecular dynamics (MD) study on CVB3 3D pol in order to address some important questions raised by experiments. First, MD simulations of F364Y and F364A were carried out to explore how these mutations at Phe364 influence active site dynamics and conformations. Second, MD simulations of wild-type and mutants were performed to discover the connection between active site dynamics and polymerase function. MD simulations reveal that the effect of mutations on active site dynamics is associated with the interaction between the structural motifs A and D in CVB3 3D pol . Interestingly, we discover that the active site state is influenced by the formation of a hydrogen bond between backbone atoms of Ala231 (in motif A) and Ala358 (in motif D), which has never been revealed before. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Asymmetric mutations in the tetrameric R67 dihydrofolate reductase reveal high tolerance to active-site substitutions.

    PubMed

    Ebert, Maximilian C C J C; Morley, Krista L; Volpato, Jordan P; Schmitzer, Andreea R; Pelletier, Joelle N

    2015-04-01

    Type II R67 dihydrofolate reductase (DHFR) is a bacterial plasmid-encoded enzyme that is intrinsically resistant to the widely-administered antibiotic trimethoprim. R67 DHFR is genetically and structurally unrelated to E. coli chromosomal DHFR and has an unusual architecture, in that four identical protomers form a single symmetrical active site tunnel that allows only one substrate binding/catalytic event at any given time. As a result, substitution of an active-site residue has as many as four distinct consequences on catalysis, constituting an atypical model of enzyme evolution. Although we previously demonstrated that no single residue of the native active site is indispensable for function, library selection here revealed a strong bias toward maintenance of two native protomers per mutated tetramer. A variety of such "half-native" tetramers were shown to procure native-like catalytic activity, with similar KM values but kcat values 5- to 33-fold lower, illustrating a high tolerance for active-site substitutions. The selected variants showed a reduced thermal stability (Tm ∼12°C lower), which appears to result from looser association of the protomers, but generally showed a marked increase in resilience to heat denaturation, recovering activity to a significantly greater extent than the variant with no active-site substitutions. Our results suggest that the presence of two native protomers in the R67 DHFR tetramer is sufficient to provide native-like catalytic rate and thus ensure cellular proliferation. © 2014 The Protein Society.

  8. Dynamics of dissolved organic matter (DOM) in a typical inland lake of the Three Gorges Reservoir area: Fluorescent properties and their implications for dissolved mercury species.

    PubMed

    Jiang, Tao; Chen, Xueshuang; Wang, Dingyong; Liang, Jian; Bai, Weiyang; Zhang, Cheng; Wang, Qilei; Wei, Shiqiang

    2018-01-15

    Dissolved organic matter (DOM) plays an important environmental and ecological role in inland aquatic systems, including lakes. In this study, using fluorescence analysis, we investigated the seasonal dynamics of DOM characteristics in Changshou Lake, which is a typical inland lake in the Three Gorges Reservoir (TGR) area. We also discuss the environmental implications of DOM for mercury (Hg) dynamics. Based on the origins of two end-members, the variations in DOM observed in this study in Changshou Lake suggest that hydrological processes (e.g., terrestrial inputs resulting from runoff and humic-like component residences) and biological activities (e.g., microbial and algae growth) are the two main principal components controlling the seasonal dynamics of DOM characteristics. Furthermore, the dynamics of dissolved Hg co-varied with variations in DOM properties, rather than with dissolved organic carbon (DOC) concentrations. This indicates that the previously reported simple correlations between DOC and Hg were not comprehensive and may lead to misunderstanding the interactions between DOM and Hg. Therefore, we recommend that when using DOM-Hg correlations to evaluate the role of DOM in the environmental fate of Hg, especially in field investigations of the spatial and temporal distribution of Hg, the properties of DOM must be taken into account. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. LASIC: Light Activated Site-Specific Conjugation of Native IgGs.

    PubMed

    Hui, James Z; Tamsen, Shereen; Song, Yang; Tsourkas, Andrew

    2015-08-19

    Numerous biological applications, from diagnostic assays to immunotherapies, rely on the use of antibody-conjugates. The efficacy of these conjugates can be significantly influenced by the site at which Immunoglobulin G (IgG) is modified. Current methods that provide control over the conjugation site, however, suffer from a number of shortfalls and often require large investments of time and cost. We have developed a novel adapter protein that, when activated by long wavelength UV light, can covalently and site-specifically label the Fc region of nearly any native, full-length IgG, including all human IgG subclasses. Labeling occurs with unprecedented efficiency and speed (>90% after 30 min), with no effect on IgG affinity. The adapter domain can be bacterially expressed and customized to contain a variety of moieties (e.g., biotin, azide, fluorophores), making reliable and efficient conjugation of antibodies widely accessible to researchers at large.

  10. ATP binding at noncatalytic sites of soluble chloroplast F1-ATPase is required for expression of the enzyme activity.

    PubMed

    Milgrom, Y M; Ehler, L L; Boyer, P D

    1990-11-05

    The F1-ATPase from chloroplasts (CF1) lacks catalytic capacity for ATP hydrolysis if ATP is not bound at noncatalytic sites. CF1 heat activated in the presence of ADP, with less than one ADP and no ATP at non-catalytic sites, shows a pronounced lag in the onset of ATP hydrolysis after exposure to 5-20 microM ATP. The onset of activity correlates well with the binding of ATP at the last two of the three noncatalytic sites. The dependence of activity on the presence of ATP at non-catalytic sites is shown at relatively low or high free Mg2+ concentrations, with or without bicarbonate as an activating anion, and when the binding of ATP at noncatalytic sites is slowed 3-4-fold by sulfate. The latent CF1 activated by dithiothreitol also requires ATP at noncatalytic sites for ATPase activity. A similar requirement by other F1-ATPases and by ATP synthases seems plausible.

  11. c-rel activates but v-rel suppresses transcription from kappa B sites.

    PubMed Central

    Inoue, J; Kerr, L D; Ransone, L J; Bengal, E; Hunter, T; Verma, I M

    1991-01-01

    We show that the product of the protooncogene c-rel is a constituent of an NF-kappa B-like complex that binds to the kappa B site originally identified in the enhancer of immunoglobulin kappa light chain gene. c-rel protein synthesized in bacteria binds to the kappa B site in a sequence-specific manner. The rel-kappa B complex can be disrupted by incubation with anti-rel antibodies. The rel protein can form oligomers. The c-rel protein can activate transcription from promoters containing kappa B sites; v-rel, on the other hand, suppresses the transcription of genes linked to kappa B sites. Thus, v-rel may interfere with the normal transcriptional machinery of the cell by acting as a dominant negative mutant. Images PMID:2023921

  12. Active-site monovalent cations revealed in a 1.55-Å-resolution hammerhead ribozyme structure.

    PubMed

    Anderson, Michael; Schultz, Eric P; Martick, Monika; Scott, William G

    2013-10-23

    We have obtained a 1.55-Å crystal structure of a hammerhead ribozyme derived from Schistosoma mansoni under conditions that permit detailed observations of Na(+) ion binding in the ribozyme's active site. At least two such Na(+) ions are observed. The first Na(+) ion binds to the N7 of G10.1 and the adjacent A9 phosphate in a manner identical with that previously observed for divalent cations. A second Na(+) ion binds to the Hoogsteen face of G12, the general base in the hammerhead cleavage reaction, thereby potentially dissipating the negative charge of the catalytically active enolate form of the nucleotide base. A potential but more ambiguous third site bridges the A9 and scissile phosphates in a manner consistent with that of previous predictions. Hammerhead ribozymes have been observed to be active in the presence of high concentrations of monovalent cations, including Na(+), but the mechanism by which monovalent cations substitute for divalent cations in hammerhead catalysis remains unclear. Our results enable us to suggest that Na(+) directly and specifically substitutes for divalent cations in the hammerhead active site. The detailed geometry of the pre-catalytic active-site complex is also revealed with a new level of precision, thanks to the quality of the electron density maps obtained from what is currently the highest-resolution ribozyme structure in the Protein Data Bank. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Computer-aided active-site-directed modeling of the Herpes Simplex Virus 1 and human thymidine kinase

    NASA Astrophysics Data System (ADS)

    Folkers, Gerd; Trumpp-Kallmeyer, Susanne; Gutbrod, Oliver; Krickl, Sabine; Fetzer, Jürgen; Keil, Günther M.

    1991-10-01

    Thymidine kinase (TK), which is induced by Herpes Simplex Virus 1 (HSV1), plays a key role in the antiviral activity of guanine derivatives such as aciclovir (ACV). In contrast, ACV shows only low affinity to the corresponding host cell enzyme. In order to define the differences in substrate binding of the two enzymes on molecular level, models for the three-dimensional (3-D) structures of the active sites of HSV1-TK and human TK were developed. The reconstruction of the active sites started from primary and secondary structure analysis of various kinases. The results were validated to homologous enzymes with known 3-D structures. The models predict that both enzymes consist of a central core β-sheet structure, connected by loops and α-helices very similar to the overall structure of other nucleotide binding enzymes. The phosphate binding is made up of a highly conserved glycine-rich loop at the N-terminus of the proteins and a conserved region at the C-terminus. The thymidine recognition site was found about 100 amino acids downstream from the phosphate binding loop. The differing substrate specificity of human and HSV1-TK can be explained by amino-acid substitutions in the homologous regions. To achieve a better understanding of the structure of the active site and how the thymidine kinase proteins interact with their substrates, the corresponding complexes of thymidine and dihydroxypropoxyguanine (DHPG) with HSV1 and human TK were built. For the docking of the guanine derivative, the X-ray structure of Elongation Factor Tu (EF-Tu), co-crystallized with guanosine diphosphate, was taken as reference. Fitting of thymidine into the active sites was done with respect to similar interactions found in thymidylate kinase. To complement the analysis of the 3-D structures of the two kinases and the substrate enzyme interactions, site-directed mutagenesis of the thymidine recognition site of HSV1-TK has been undertaken, changing Asp162 in the thymidine recognition site

  14. Evidence for histidine in the active sites of ficin and stem-bromelain

    PubMed Central

    Husain, S. S.; Lowe, G.

    1968-01-01

    1. Ficin and stem-bromelain are irreversibly inhibited by 1,3-dibromoacetone, a reagent designed to react first with the active-site cysteine residue and subsequently with a second nucleophile. Evidence is presented that establishes that a histidine residue is within a 5Å locus of the active-site cysteine residue in both enzymes. The histidine residue in both enzymes is alkylated at N-1 by dibromoacetone. It is suggested that, as with papain, the thiol and imidazole groups act in concert in the hydrolysis of substrates by these enzymes. 2. The inhibition of thiol-subtilisin with 1,3-dibromoacetone is shown to be due to the alkylation of a cysteine residue only. PMID:5722692

  15. Analysis on biomass and productivity of epilithic algae and their relations to environmental factors in the Gufu River basin, Three Gorges Reservoir area, China.

    PubMed

    Ge, Jiwen; Wu, Shuyuan; Touré, Dado; Cheng, Lamei; Miao, Wenjie; Cao, Huafen; Pan, Xiaoying; Li, Jianfeng; Yao, Minmin; Feng, Liang

    2017-12-01

    The main purpose of this study conducted from August 2010 was to find biomass and productivity of epilithic algae and their relations to environmental factors and try to explore the restrictive factors affecting the growth of algae in the Gufu River, the one of the branches of Xiangxi River located in the Three Gorges Reservoir of the Yangtze River, Hubei Province, Central China. An improved method of in situ primary productivity measurement was utilized to estimate the primary production of the epilithic algae. It was shown that in rivers, lakes, and reservoirs, algae are the main primary producers and have a central role in the ecosystem. Chlorophyll a concentration and ash-free dry mass (AFDM) were estimated for epilithic algae of the Gufu River basin in Three Gorges Reservoir area. Environmental factors in the Gufu River ecosystem highlighted differences in periphyton chlorophyll a ranging from 1.49 mg m -2 (origin) to 69.58 mg m -2 (terminal point). The minimum and maximum gross primary productivity of epilithic algae were 96.12 and 1439.89 mg C m -2  day -1 , respectively. The mean net primary productivity was 290.24 mg C m -2  day -1 . The mean autotrophic index (AFDM:chlorophyll a) was 407.40. The net primary productivity, community respiration ratio (P/R ratio) ranged from 0.98 to 9.25 with a mean of 2.76, showed that autotrophic productivity was dominant in the river. Relationship between physicochemical characteristics and biomass was discussed through cluster and stepwise regression analysis which indicated that altitude, total nitrogen (TN), NO 3 - -N, and NH 4 + -N were significant environmental factors affecting the biomass of epilithic algae. However, a negative logarithmic relationship between altitude and the chlorophyll a of epilithic algae was high. The results also highlighted the importance of epilithic algae in maintaining the Gufu River basin ecosystems health.

  16. Crystal structure of an avian influenza polymerase PA[subscript N] reveals an endonuclease active site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yuan, Puwei; Bartlam, Mark; Lou, Zhiyong

    2009-11-10

    The heterotrimeric influenza virus polymerase, containing the PA, PB1 and PB2 proteins, catalyses viral RNA replication and transcription in the nucleus of infected cells. PB1 holds the polymerase active site and reportedly harbours endonuclease activity, whereas PB2 is responsible for cap binding. The PA amino terminus is understood to be the major functional part of the PA protein and has been implicated in several roles, including endonuclease and protease activities as well as viral RNA/complementary RNA promoter binding. Here we report the 2.2 angstrom (A) crystal structure of the N-terminal 197 residues of PA, termed PA(N), from an avian influenzamore » H5N1 virus. The PA(N) structure has an alpha/beta architecture and reveals a bound magnesium ion coordinated by a motif similar to the (P)DX(N)(D/E)XK motif characteristic of many endonucleases. Structural comparisons and mutagenesis analysis of the motif identified in PA(N) provide further evidence that PA(N) holds an endonuclease active site. Furthermore, functional analysis with in vivo ribonucleoprotein reconstitution and direct in vitro endonuclease assays strongly suggest that PA(N) holds the endonuclease active site and has critical roles in endonuclease activity of the influenza virus polymerase, rather than PB1. The high conservation of this endonuclease active site among influenza strains indicates that PA(N) is an important target for the design of new anti-influenza therapeutics.« less

  17. Discovery of HDAC Inhibitors That Lack an Active Site Zn(2+)-Binding Functional Group.

    PubMed

    Vickers, Chris J; Olsen, Christian A; Leman, Luke J; Ghadiri, M Reza

    2012-06-14

    Natural and synthetic histone deacetylase (HDAC) inhibitors generally derive their strong binding affinity and high potency from a key functional group that binds to the Zn(2+) ion within the enzyme active site. However, this feature is also thought to carry the potential liability of undesirable off-target interactions with other metalloenzymes. As a step toward mitigating this issue, here, we describe the design, synthesis, and structure-activity characterizations of cyclic α3β-tetrapeptide HDAC inhibitors that lack the presumed indispensable Zn(2+)-binding group. The lead compounds (e.g., 15 and 26) display good potency against class 1 HDACs and are active in tissue culture against various human cancer cell lines. Importantly, enzymological analysis of 26 indicates that the cyclic α3β-tetrapeptide is a fast-on/off competitive inhibitor of HDACs 1-3 with K i values of 49, 33, and 37 nM, respectively. Our proof of principle study supports the idea that novel classes of HDAC inhibitors, which interact at the active-site opening, but not with the active site Zn(2+), can have potential in drug design.

  18. Structural insight into the active site of mushroom tyrosinase using phenylbenzoic acid derivatives.

    PubMed

    Oyama, Takahiro; Yoshimori, Atsushi; Takahashi, Satoshi; Yamamoto, Tetsuya; Sato, Akira; Kamiya, Takanori; Abe, Hideaki; Abe, Takehiko; Tanuma, Sei-Ichi

    2017-07-01

    So far, many inhibitors of tyrosinase have been discovered for cosmetic and clinical agents. However, the molecular mechanisms underlying the inhibition in the active site of tyrosinase have not been well understood. To explore this problem, we examined here the inhibitory effects of 4'-hydroxylation and methoxylation of phenylbenzoic acid (PBA) isomers, which have a unique scaffold to inhibit mushroom tyrosinase. The inhibitory effect of 3-PBA, which has the most potent inhibitory activity among the isomers, was slightly decreased by 4'-hydroxylation and further decreased by 4'-methoxylation against mushroom tyrosinase. Surprisingly, 4'-hydroxylation but not methoxylation of 2-PBA appeared inhibitory activity. On the other hand, both 4'-hydroxylation and methoxylation of 4-PBA increased the inhibitory activity against mushroom tyrosinase. In silico docking analyses using the crystallographic structure of mushroom tyrosinase indicated that the carboxylic acid or 4'-hydroxyl group of PBA derivatives could chelate with cupric ions in the active site of mushroom tyrosinase, and that the interactions of Asn260 and Phe264 in the active site with the adequate-angled biphenyl group are involved in the inhibitory activities of the modified PBAs, by parallel and T-shaped π-π interactions, respectively. Furthermore, Arg268 could fix the angle of the aromatic ring of Phe264, and Val248 is supposed to interact with the inhibitors as a hydrophobic manner. These results may enhance the structural insight into mushroom tyrosinase for the creation of novel tyrosinase inhibitors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Nature of the Active Sites for CO Reduction on Copper Nanoparticles; Suggestions for Optimizing Performance.

    PubMed

    Cheng, Tao; Xiao, Hai; Goddard, William A

    2017-08-30

    Recent experiments show that the grain boundaries (GBs) of copper nanoparticles (NPs) lead to an outstanding performance in reducing CO 2 and CO to alcohol products. We report here multiscale simulations that simulate experimental synthesis conditions to predict the structure of a 10 nm Cu NP (158 555 atoms). To identify active sites, we first predict the CO binding at a large number of sites and select four exhibiting CO binding stronger than the (211) step surface. Then, we predict the formation energy of the *OCCOH intermediate as a descriptor for C-C coupling, identifying two active sites, both of which have an under-coordinated surface square site adjacent to a subsurface stacking fault. We then propose a periodic Cu surface (4 by 4 supercell) with a similar site that substantially decreases the formation energy of *OCCOH, by 0.14 eV.

  20. Wobble pairs of the HDV ribozyme play specific roles in stabilization of active site dynamics.

    PubMed

    Sripathi, Kamali N; Banáš, Pavel; Réblová, Kamila; Šponer, Jiří; Otyepka, Michal; Walter, Nils G

    2015-02-28

    The hepatitis delta virus (HDV) is the only known human pathogen whose genome contains a catalytic RNA motif (ribozyme). The overall architecture of the HDV ribozyme is that of a double-nested pseudoknot, with two GU pairs flanking the active site. Although extensive studies have shown that mutation of either wobble results in decreased catalytic activity, little work has focused on linking these mutations to specific structural effects on catalytic fitness. Here we use molecular dynamics simulations based on an activated structure to probe the active site dynamics as a result of wobble pair mutations. In both wild-type and mutant ribozymes, the in-line fitness of the active site (as a measure of catalytic proficiency) strongly depends on the presence of a C75(N3H3+)N1(O5') hydrogen bond, which positions C75 as the general acid for the reaction. Our mutational analyses show that each GU wobble supports catalytically fit conformations in distinct ways; the reverse G25U20 wobble promotes high in-line fitness, high occupancy of the C75(N3H3+)G1(O5') general-acid hydrogen bond and stabilization of the G1U37 wobble, while the G1U37 wobble acts more locally by stabilizing high in-line fitness and the C75(N3H3+)G1(O5') hydrogen bond. We also find that stable type I A-minor and P1.1 hydrogen bonding above and below the active site, respectively, prevent local structural disorder from spreading and disrupting global conformation. Taken together, our results define specific, often redundant architectural roles for several structural motifs of the HDV ribozyme active site, expanding the known roles of these motifs within all HDV-like ribozymes and other structured RNAs.

  1. Wobble Pairs of the HDV Ribozyme Play Specific Roles in Stabilization of Active Site Dynamics

    PubMed Central

    Sripathi, Kamali N.; Banáš, Pavel; Reblova, Kamila; Šponer, Jiři; Otyepka, Michal

    2015-01-01

    The hepatitis delta virus (HDV) is the only known human pathogen whose genome contains a catalytic RNA motif (ribozyme). The overall architecture of the HDV ribozyme is that of a double-nested pseudoknot, with two GU pairs flanking the active site. Although extensive studies have shown that mutation of either wobble results in decreased catalytic activity, little work has focused on linking these mutations to specific structural effects on catalytic fitness. Here we use molecular dynamics simulations based on an activated structure to probe the active site dynamics as a result of wobble pair mutations. In both wild-type and mutant ribozymes, the in-line fitness of the active site (as a measure of catalytic proficiency) strongly depends on the presence of a C75(N3H3+)N1(O5′) hydrogen bond, which positions C75 as the general acid for the reaction. Our mutational analyses show that each GU wobble supports catalytically fit conformations in distinct ways; the reverse G25U20 wobble promotes high in-line fitness, high occupancy of the C75(N3H3+)G1(O5′) general-acid hydrogen bond and stabilization of the G1U37 wobble, while the G1U37 wobble acts more locally by stabilizing high in-line fitness and the C75(N3H3+)G1(O5′) hydrogen bond. We also find that stable type I A-minor and P1.1 hydrogen bonding above and below the active site, respectively, prevent local structural disorder from spreading and disrupting global conformation. Taken together, our results define specific, often redundant architectural roles for several structural motifs of the HDV ribozyme active site, expanding the known roles of these motifs within all HDV-like ribozymes and other structured RNAs. PMID:25631765

  2. Severe situation of rural nonpoint source pollution and efficient utilization of agricultural wastes in the Three Gorges Reservoir Area.

    PubMed

    Zhang, Tong; Ni, Jiupai; Xie, Deti

    2015-11-01

    Rural nonpoint source (NPS) pollution caused by agricultural wastes has become increasingly serious in the Three Gorges Reservoir Area (TGRA), significantly affecting the reservoir water quality. The grim situation of rural NPS pollution in the TGRA indicated that agrochemicals (chemical fertilizer and pesticide) were currently the highest contributor of rural NPS pollution (50.38%). The harmless disposal rates of livestock excrement, crop straws, rural domestic refuse, and sewage also cause severe water pollution. More importantly, the backward agricultural economy and the poor environmental awareness of farmers in the hinterland of the TGRA contribute to high levels of rural NPS pollution. Over the past decade, researchers and the local people have carried out various successful studies and practices to realize the effective control of rural NPS pollution by efficiently utilizing agricultural wastes in the TGRA, including agricultural waste biogas-oriented utilization, crop straw gasification, decentralized land treatment of livestock excrement technology, and crop straw modification. These technologies have greatly increased the renewable resource utilization of agricultural wastes and improved water quality and ecological environment in the TGRA.

  3. Quantitative functional characterization of conserved molecular interactions in the active site of mannitol 2-dehydrogenase

    PubMed Central

    Lucas, James E; Siegel, Justin B

    2015-01-01

    Enzyme active site residues are often highly conserved, indicating a significant role in function. In this study we quantitate the functional contribution for all conserved molecular interactions occurring within a Michaelis complex for mannitol 2-dehydrogenase derived from Pseudomonas fluorescens (pfMDH). Through systematic mutagenesis of active site residues, we reveal that the molecular interactions in pfMDH mediated by highly conserved residues not directly involved in reaction chemistry can be as important to catalysis as those directly involved in the reaction chemistry. This quantitative analysis of the molecular interactions within the pfMDH active site provides direct insight into the functional role of each molecular interaction, several of which were unexpected based on canonical sequence conservation and structural analyses. PMID:25752240

  4. A gratuitous β-Lactamase inducer uncovers hidden active site dynamics of the Staphylococcus aureus BlaR1 sensor domain.

    PubMed

    Frederick, Thomas E; Peng, Jeffrey W

    2018-01-01

    Increasing evidence shows that active sites of proteins have non-trivial conformational dynamics. These dynamics include active site residues sampling different local conformations that allow for multiple, and possibly novel, inhibitor binding poses. Yet, active site dynamics garner only marginal attention in most inhibitor design efforts and exert little influence on synthesis strategies. This is partly because synthesis requires a level of atomic structural detail that is frequently missing in current characterizations of conformational dynamics. In particular, while the identity of the mobile protein residues may be clear, the specific conformations they sample remain obscure. Here, we show how an appropriate choice of ligand can significantly sharpen our abilities to describe the interconverting binding poses (conformations) of protein active sites. Specifically, we show how 2-(2'-carboxyphenyl)-benzoyl-6-aminopenicillanic acid (CBAP) exposes otherwise hidden dynamics of a protein active site that binds β-lactam antibiotics. When CBAP acylates (binds) the active site serine of the β-lactam sensor domain of BlaR1 (BlaRS), it shifts the time scale of the active site dynamics to the slow exchange regime. Slow exchange enables direct characterization of inter-converting protein and bound ligand conformations using NMR methods. These methods include chemical shift analysis, 2-d exchange spectroscopy, off-resonance ROESY of the bound ligand, and reduced spectral density mapping. The active site architecture of BlaRS is shared by many β-lactamases of therapeutic interest, suggesting CBAP could expose functional motions in other β-lactam binding proteins. More broadly, CBAP highlights the utility of identifying chemical probes common to structurally homologous proteins to better expose functional motions of active sites.

  5. Target-classification approach applied to active UXO sites

    NASA Astrophysics Data System (ADS)

    Shubitidze, F.; Fernández, J. P.; Shamatava, Irma; Barrowes, B. E.; O'Neill, K.

    2013-06-01

    This study is designed to illustrate the discrimination performance at two UXO active sites (Oklahoma's Fort Sill and the Massachusetts Military Reservation) of a set of advanced electromagnetic induction (EMI) inversion/discrimination models which include the orthonormalized volume magnetic source (ONVMS), joint diagonalization (JD), and differential evolution (DE) approaches and whose power and flexibility greatly exceed those of the simple dipole model. The Fort Sill site is highly contaminated by a mix of the following types of munitions: 37-mm target practice tracers, 60-mm illumination mortars, 75-mm and 4.5'' projectiles, 3.5'', 2.36'', and LAAW rockets, antitank mine fuzes with and without hex nuts, practice MK2 and M67 grenades, 2.5'' ballistic windshields, M2A1-mines with/without bases, M19-14 time fuzes, and 40-mm practice grenades with/without cartridges. The site at the MMR site contains targets of yet different sizes. In this work we apply our models to EMI data collected using the MetalMapper (MM) and 2 × 2 TEMTADS sensors. The data for each anomaly are inverted to extract estimates of the extrinsic and intrinsic parameters associated with each buried target. (The latter include the total volume magnetic source or NVMS, which relates to size, shape, and material properties; the former includes location, depth, and orientation). The estimated intrinsic parameters are then used for classification performed via library matching and the use of statistical classification algorithms; this process yielded prioritized dig-lists that were submitted to the Institute for Defense Analyses (IDA) for independent scoring. The models' classification performance is illustrated and assessed based on these independent evaluations.

  6. Cyanide does more to inhibit heme enzymes, than merely serving as an active-site ligand.

    PubMed

    Parashar, Abhinav; Venkatachalam, Avanthika; Gideon, Daniel Andrew; Manoj, Kelath Murali

    2014-12-12

    The toxicity of cyanide is hitherto attributed to its ability to bind to heme proteins' active site and thereby inhibit their activity. It is shown herein that the long-held interpretation is inadequate to explain several observations in heme-enzyme reaction systems. Generation of cyanide-based diffusible radicals in heme-enzyme reaction milieu could shunt electron transfers (by non-active site processes), and thus be detrimental to the efficiency of oxidative outcomes. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. An evaluation of neonicotinoids' potential to inhibit human cholinesterases: Protein-ligand docking and interaction profiling studies.

    PubMed

    Teralı, Kerem

    2018-06-16

    Many so-called neuroactive insecticides target invertebrate neurotransmitter systems, including the cholinergic system. With their relatively low toxicity to vertebrates, neonicotinoids represent a new class of neuroactive insecticides that bind to nicotinic receptors for acetylcholine in the insect central nervous system and result in paralysis and eventual death due to receptor overstimulation. On the understanding that, today, cholinesterase inhibitors are used to obtain the symptomatic relief of Alzheimer disease (AD), the aforementioned direct cholinomimetic action of neonicotinoids could, perhaps, confer anti-AD drug-like attributes to these compounds. It is shown here, using protein-ligand docking and interaction profiling, that neonicotinoids penetrate deep into the active-site gorge of both acetylcholinesterase and butyrylcholinesterase and that they form relatively strong noncovalent bonds with multiple critical residues that normally bind/hydrolyze choline esters. With their gorge-spanning shape and dual-binding specificity, neonicotinoids (first-generation compounds in particular) represent promising leads for the development of reversible, mixed-type cholinesterase inhibitors in the fight against AD. Copyright © 2018 Elsevier Inc. All rights reserved.

  8. Quantum delocalization of protons in the hydrogen-bond network of an enzyme active site

    PubMed Central

    Wang, Lu; Fried, Stephen D.; Boxer, Steven G.; Markland, Thomas E.

    2014-01-01

    Enzymes use protein architectures to create highly specialized structural motifs that can greatly enhance the rates of complex chemical transformations. Here, we use experiments, combined with ab initio simulations that exactly include nuclear quantum effects, to show that a triad of strongly hydrogen-bonded tyrosine residues within the active site of the enzyme ketosteroid isomerase (KSI) facilitates quantum proton delocalization. This delocalization dramatically stabilizes the deprotonation of an active-site tyrosine residue, resulting in a very large isotope effect on its acidity. When an intermediate analog is docked, it is incorporated into the hydrogen-bond network, giving rise to extended quantum proton delocalization in the active site. These results shed light on the role of nuclear quantum effects in the hydrogen-bond network that stabilizes the reactive intermediate of KSI, and the behavior of protons in biological systems containing strong hydrogen bonds. PMID:25503367

  9. Evaluating the Substrate Selectivity of Alkyladenine DNA Glycosylase: The Synergistic Interplay of Active Site Flexibility and Water Reorganization.

    PubMed

    Lenz, Stefan A P; Wetmore, Stacey D

    2016-02-09

    Human alkyladenine DNA glycosylase (AAG) functions as part of the base excision repair (BER) pathway by cleaving the N-glycosidic bond that connects nucleobases to the sugar-phosphate backbone in DNA. AAG targets a range of structurally diverse purine lesions using nonspecific DNA-protein π-π interactions. Nevertheless, the enzyme discriminates against the natural purines and is inhibited by pyrimidine lesions. This study uses molecular dynamics simulations and seven different neutral or charged substrates, inhibitors, or canonical purines to probe how the bound nucleotide affects the conformation of the AAG active site, and the role of active site residues in dictating substrate selectivity. The neutral substrates form a common DNA-protein hydrogen bond, which results in a consistent active site conformation that maximizes π-π interactions between the aromatic residues and the nucleobase required for catalysis. Nevertheless, subtle differences in DNA-enzyme contacts for different neutral substrates explain observed differential catalytic efficiencies. In contrast, the exocyclic amino groups of the natural purines clash with active site residues, which leads to catalytically incompetent DNA-enzyme complexes due to significant reorganization of active site water. Specifically, water resides between the A nucleobase and the active site aromatic amino acids required for catalysis, while a shift in the position of the general base (E125) repositions (potentially nucleophilic) water away from G. Despite sharing common amino groups, the methyl substituents in cationic purine lesions (3MeA and 7MeG) exhibit repulsion with active site residues, which repositions the damaged bases in the active site in a manner that promotes their excision. Overall, we provide a structural explanation for the diverse yet discriminatory substrate selectivity of AAG and rationalize key kinetic data available for the enzyme. Specifically, our results highlight the complex interplay of many

  10. Parkin-phosphoubiquitin complex reveals cryptic ubiquitin-binding site required for RBR ligase activity.

    PubMed

    Kumar, Atul; Chaugule, Viduth K; Condos, Tara E C; Barber, Kathryn R; Johnson, Clare; Toth, Rachel; Sundaramoorthy, Ramasubramanian; Knebel, Axel; Shaw, Gary S; Walden, Helen

    2017-05-01

    RING-between-RING (RBR) E3 ligases are a class of ubiquitin ligases distinct from RING or HECT E3 ligases. An important RBR ligase is Parkin, mutations in which lead to early-onset hereditary Parkinsonism. Parkin and other RBR ligases share a catalytic RBR module but are usually autoinhibited and activated via distinct mechanisms. Recent insights into Parkin regulation predict large, unknown conformational changes during Parkin activation. However, current data on active RBR ligases reflect the absence of regulatory domains. Therefore, it remains unclear how individual RBR ligases are activated, and whether they share a common mechanism. We now report the crystal structure of a human Parkin-phosphoubiquitin complex, which shows that phosphoubiquitin binding induces movement in the 'in-between RING' (IBR) domain to reveal a cryptic ubiquitin-binding site. Mutation of this site negatively affects Parkin's activity. Furthermore, ubiquitin binding promotes cooperation between Parkin molecules, which suggests a role for interdomain association in the RBR ligase mechanism.

  11. The specific interaction of the photosensitizer methylene blue with acetylcholinesterase provides a model system for studying the molecular consequences of photodynamic therapy.

    PubMed

    Silman, Israel; Roth, Esther; Paz, Aviv; Triquigneaux, Mathilde M; Ehrenshaft, Marilyn; Xu, Yechun; Shnyrov, Valery L; Sussman, Joel L; Deterding, Leesa J; Ashani, Yacov; Mason, Ronald P; Weiner, Lev

    2013-03-25

    The photosensitizer, methylene blue (MB), generates singlet oxygen ((1)O2) that irreversibly inhibits Torpedo californica acetylcholinesterase (TcAChE). In the dark MB inhibits reversibly, binding being accompanied by a bathochromic shift that can be used to show its displacement by other reversible inhibitors binding to the catalytic 'anionic' subsite (CAS), the peripheral 'anionic' subsite (PAS), or bridging them. Data concerning both reversible and irreversible inhibition are here reviewed. MB protects TcAChE from thermal denaturation, and differential scanning calorimetry reveals a ~8 °C increase in the denaturation temperature. The crystal structure of the MB/TcAChE complex reveals a single MB stacked against W279 in the PAS, pointing down the gorge towards the CAS. The intrinsic fluorescence of the irreversibly inhibited enzyme displays new emission bands that can be ascribed to N'-formylkynurenine (NFK); this was indeed confirmed using anti-NFK antibodies. Mass spectroscopy revealed that two Trp residues, Trp84 in the CAS, and Trp279 in the PAS, were the only Trp residues, out of a total of 14, significantly modified by photo-oxidation, both being converted to NFK. In the presence of competitive inhibitors that displace MB from the gorge, their modification is completely prevented. Thus, photo-oxidative damage caused by MB involves targeted release of (1)O2 by the bound photosensitizer within the aqueous milieu of the active-site gorge. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  12. Regulation of Dpp activity by tissue-specific cleavage of an upstream site within the prodomain

    PubMed Central

    Sopory, Shailaja; Kwon, Sunjong; Wehrli, Marcel; Christian, Jan L.

    2010-01-01

    BMP4 is synthesized as an inactive precursor that is cleaved at two sites during maturation: initially at a site (S1) adjacent to the ligand domain, and then at an upstream site (S2) within the prodomain. Cleavage at the second site regulates the stability of mature BMP4 and this in turn influences its signaling intensity and range of action. The Drosophila ortholog of BMP4, Dpp, functions as a long- or short-range signaling molecule in the wing disc or embryonic midgut, respectively but mechanisms that differentially regulate its bioactivity in these tissues have not been explored. In the current studies we demonstrate, by dpp mutant rescue, that cleavage at the S2 site of proDpp is required for development of the wing and leg imaginal discs, whereas cleavage at the S1 site is sufficient to rescue Dpp function in the midgut. Both the S1 and S2 site of proDpp are cleaved in the wing disc, and S2-cleavage is essential to generate sufficient ligand to exceed the threshold for pMAD activation at both short- and long-range in most cells. By contrast, proDpp is cleaved at the S1 site alone in the embryonic mesoderm and this generates sufficient ligand to activate physiological target genes in neighboring cells. These studies provide the first biochemical and genetic evidence that that selective cleavage of the S2 site of proDPP provides a tissue-specific mechanism for regulating Dpp activity, and that differential cleavage can contribute to, but is not an absolute determinant of signaling range. PMID:20659445

  13. A three-dimensional model of mammalian tyrosinase active site accounting for loss of function mutations.

    PubMed

    Schweikardt, Thorsten; Olivares, Concepción; Solano, Francisco; Jaenicke, Elmar; García-Borrón, José Carlos; Decker, Heinz

    2007-10-01

    Tyrosinases are the first and rate-limiting enzymes in the synthesis of melanin pigments responsible for colouring hair, skin and eyes. Mutation of tyrosinases often decreases melanin production resulting in albinism, but the effects are not always understood at the molecular level. Homology modelling of mouse tyrosinase based on recently published crystal structures of non-mammalian tyrosinases provides an active site model accounting for loss-of-function mutations. According to the model, the copper-binding histidines are located in a helix bundle comprising four densely packed helices. A loop containing residues M374, S375 and V377 connects the CuA and CuB centres, with the peptide oxygens of M374 and V377 serving as hydrogen acceptors for the NH-groups of the imidazole rings of the copper-binding His367 and His180. Therefore, this loop is essential for the stability of the active site architecture. A double substitution (374)MS(375) --> (374)GG(375) or a single M374G mutation lead to a local perturbation of the protein matrix at the active site affecting the orientation of the H367 side chain, that may be unable to bind CuB reliably, resulting in loss of activity. The model also accounts for loss of function in two naturally occurring albino mutations, S380P and V393F. The hydroxyl group in S380 contributes to the correct orientation of M374, and the substitution of V393 for a bulkier phenylalanine sterically impedes correct side chain packing at the active site. Therefore, our model explains the mechanistic necessity for conservation of not only active site histidines but also adjacent amino acids in tyrosinase.

  14. Evaluation of physical activity web sites for use of behavior change theories.

    PubMed

    Doshi, Amol; Patrick, Kevin; Sallis, James F; Calfas, Karen

    2003-01-01

    Physical activity (PA) Web sites were assessed for their use of behavior change theories, including constructs of the health belief model, Transtheoretical Model, social cognitive theory, and the theory of reasoned action and planned behavior. An evaluation template for assessing PA Web sites was developed, and content validity and interrater reliability were demonstrated. Two independent raters evaluated 24 PA Web sites. Web sites varied widely in application of theory-based constructs, ranging from 5 to 48 on a 100-point scale. The most common intervention strategies were general information, social support, and realistic goal areas. Coverage of theory-based strategies was low, varying from 26% for social cognitive theory to 39% for health belief model. Overall, PA Web sites provided little assessment, feedback, or individually tailored assistance for users. They were unable to substantially tailor the on-line experience for users at different stages of change or different demographic characteristics.

  15. Active-Site Plasticity Is Essential to Carbapenem Hydrolysis by OXA-58 Class D β-Lactamase of Acinetobacter baumannii.

    PubMed

    Pratap, Shivendra; Katiki, Madhusudhanarao; Gill, Preet; Kumar, Pravindra; Golemi-Kotra, Dasantila

    2016-01-01

    Carbapenem-hydrolyzing class D β-lactamases (CHDLs) are a subgroup of class D β-lactamases, which are enzymes that hydrolyze β-lactams. They have attracted interest due to the emergence of multidrug-resistant Acinetobacter baumannii, which is not responsive to treatment with carbapenems, the usual antibiotics of choice for this bacterium. Unlike other class D β-lactamases, these enzymes efficiently hydrolyze carbapenem antibiotics. To explore the structural requirements for the catalysis of carbapenems by these enzymes, we determined the crystal structure of the OXA-58 CHDL of A. baumannii following acylation of its active-site serine by a 6α-hydroxymethyl penicillin derivative that is a structural mimetic for a carbapenem. In addition, several point mutation variants of the active site of OXA-58, as identified by the crystal structure analysis, were characterized kinetically. These combined studies confirm the mechanistic relevance of a hydrophobic bridge formed over the active site. This structural feature is suggested to stabilize the hydrolysis-productive acyl-enzyme species formed from the carbapenem substrates of this enzyme. Furthermore, our structural studies provide strong evidence that the hydroxyethyl group of carbapenems samples different orientations in the active sites of CHDLs, and the optimum orientation for catalysis depends on the topology of the active site allowing proper closure of the active site. We propose that CHDLs use the plasticity of the active site to drive the mechanism of carbapenem hydrolysis toward efficiency. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  16. Dynamic asymmetry and the role of the conserved active-site thiol in rabbit muscle creatine kinase.

    PubMed

    Londergan, Casey H; Baskin, Rachel; Bischak, Connor G; Hoffman, Kevin W; Snead, David M; Reynoso, Christopher

    2015-01-13

    Symmetric and asymmetric crystal structures of the apo and transition state analogue forms, respectively, of the dimeric rabbit muscle creatine kinase have invoked an "induced fit" explanation for asymmetry between the two subunits and their active sites. However, previously reported thiol reactivity studies at the dual active-site cysteine 283 residues suggest a more latent asymmetry between the two subunits. The role of that highly conserved active-site cysteine has also not been clearly determined. In this work, the S-H vibrations of Cys283 were observed in the unmodified MM isoform enzyme via Raman scattering, and then one and both Cys283 residues in the same dimeric enzyme were modified to covalently attach a cyano group that reports on the active-site environment via its infrared CN stretching absorption band while maintaining the catalytic activity of the enzyme. Unmodified and Cys283-modified enzymes were investigated in the apo and transition state analogue forms of the enzyme. The narrow and invariant S-H vibrational bands report a homogeneous environment for the unmodified active-site cysteines, indicating that their thiols are hydrogen bonded to the same H-bond acceptor in the presence and absence of the substrate. The S-H peak persists at all physiologically relevant pH's, indicating that Cys283 is protonated at all pH's relevant to enzymatic activity. Molecular dynamics simulations identify the S-H hydrogen bond acceptor as a single, long-resident water molecule and suggest that the role of the conserved yet catalytically unnecessary thiol may be to dynamically rigidify that part of the active site through specific H-bonding to water. The asymmetric and broad CN stretching bands from the CN-modified Cys283 suggest an asymmetric structure in the apo form of the enzyme in which there is a dynamic exchange between spectral subpopulations associated with water-exposed and water-excluded probe environments. Molecular dynamics simulations indicate a

  17. 78 FR 8190 - Commercial Wind Leasing and Site Assessment Activities on the Atlantic Outer Continental Shelf...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-05

    ...] Commercial Wind Leasing and Site Assessment Activities on the Atlantic Outer Continental Shelf (OCS) Offshore... Assessment (EA) for Commercial Wind Leasing and Site Assessment Activities on the OCS Offshore North Carolina... INFORMATION: Background: On December 13, 2012, BOEM published the Notice in the Federal Register (77 FR 74218...

  18. Altered binding of thioflavin t to the peripheral anionic site of acetylcholinesterase after phosphorylation of the active site by chlorpyrifos oxon or dichlorvos

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sultatos, L.G.; Kaushik, R.

    2008-08-01

    The peripheral anionic site of acetylcholinesterase, when occupied by a ligand, is known to modulate reaction rates at the active site of this important enzyme. The current report utilized the peripheral anionic site specific fluorogenic probe thioflavin t to determine if the organophosphates chlorpyrifos oxon and dichlorvos bind to the peripheral anionic site of human recombinant acetylcholinesterase, since certain organophosphates display concentration-dependent kinetics when inhibiting this enzyme. Incubation of 3 nM acetylcholinesterase active sites with 50 nM or 2000 nM inhibitor altered both the B{sub max} and K{sub d} for thioflavin t binding to the peripheral anionic site. However, thesemore » changes resulted from phosphorylation of Ser203 since increasing either inhibitor from 50 nM to 2000 nM did not alter further thioflavin t binding kinetics. Moreover, the organophosphate-induced decrease in B{sub max} did not represent an actual reduction in binding sites, but instead likely resulted from conformational interactions between the acylation and peripheral anionic sites that led to a decrease in the rigidity of bound thioflavin t. A drop in fluorescence quantum yield, leading to an apparent decrease in B{sub max}, would accompany the decreased rigidity of bound thioflavin t molecules. The organophosphate-induced alterations in K{sub d} represented changes in binding affinity of thioflavin t, with diethylphosphorylation of Ser203 increasing K{sub d}, and dimethylphosphorylation of Ser203 decreasing K{sub d}. These results indicate that chlorpyrifos oxon and dichlorvos do not bind directly to the peripheral anionic site of acetylcholinesterase, but can affect binding to that site through phosphorylation of Ser203.« less

  19. Cyanide does more to inhibit heme enzymes, than merely serving as an active-site ligand

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Parashar, Abhinav; Venkatachalam, Avanthika; Gideon, Daniel Andrew

    Highlights: • Cyanide (CN) is a well-studied toxic principle, known to inhibit heme-enzymes. • Inhibition is supposed to result from CN binding at the active site as a ligand. • Diverse heme enzymes’ CN inhibition profiles challenge prevailing mechanism. • Poor binding efficiency of CN at low enzyme concentrations and ligand pressures. • CN-based diffusible radicals cause ‘non-productive electron transfers’ (inhibition). - Abstract: The toxicity of cyanide is hitherto attributed to its ability to bind to heme proteins’ active site and thereby inhibit their activity. It is shown herein that the long-held interpretation is inadequate to explain several observations inmore » heme-enzyme reaction systems. Generation of cyanide-based diffusible radicals in heme-enzyme reaction milieu could shunt electron transfers (by non-active site processes), and thus be detrimental to the efficiency of oxidative outcomes.« less

  20. A facile reflux procedure to increase active surface sites form highly active and durable supported palladium@platinum bimetallic nanodendrites

    NASA Astrophysics Data System (ADS)

    Wang, Qin; Li, Yingjun; Liu, Baocang; Xu, Guangran; Zhang, Geng; Zhao, Qi; Zhang, Jun

    2015-11-01

    A series of well-dispersed bimetallic Pd@Pt nanodendrites uniformly supported on XC-72 carbon black are fabricated by using different capping agents. These capping agents are essential for the branched morphology control. However, the surfactant adsorbed on the nanodendrites surface blocks the access of reactant molecules to the active surface sites, and the catalytic activities of these bimetallic nanodendrites are significantly restricted. Herein, a facile reflux procedure to effectively remove the capping agent molecules without significantly affecting their sizes is reported for activating supported nanocatalysts. More significantly, the structure and morphology of the nanodendrites can also be retained, enhancing the numbers of active surface sites, catalytic activity and stability toward methanol and ethanol electro-oxidation reactions. The as-obtained hot water reflux-treated Pd@Pt/C catalyst manifests superior catalytic activity and stability both in terms of surface and mass specific activities, as compared to the untreated catalysts and the commercial Pt/C and Pd/C catalysts. We anticipate that this effective and facile removal method has more general applicability to highly active nanocatalysts prepared with various surfactants, and should lead to improvements in environmental protection and energy production.

  1. Site-specific Interaction Mapping of Phosphorylated Ubiquitin to Uncover Parkin Activation*♦

    PubMed Central

    Yamano, Koji; Queliconi, Bruno B.; Koyano, Fumika; Saeki, Yasushi; Hirokawa, Takatsugu; Tanaka, Keiji; Matsuda, Noriyuki

    2015-01-01

    Damaged mitochondria are eliminated through autophagy machinery. A cytosolic E3 ubiquitin ligase Parkin, a gene product mutated in familial Parkinsonism, is essential for this pathway. Recent progress has revealed that phosphorylation of both Parkin and ubiquitin at Ser65 by PINK1 are crucial for activation and recruitment of Parkin to the damaged mitochondria. However, the mechanism by which phosphorylated ubiquitin associates with and activates phosphorylated Parkin E3 ligase activity remains largely unknown. Here, we analyze interactions between phosphorylated forms of both Parkin and ubiquitin at a spatial resolution of the amino acid residue by site-specific photo-crosslinking. We reveal that the in-between-RING (IBR) domain along with RING1 domain of Parkin preferentially binds to ubiquitin in a phosphorylation-dependent manner. Furthermore, another approach, the Fluoppi (fluorescent-based technology detecting protein-protein interaction) assay, also showed that pathogenic mutations in these domains blocked interactions with phosphomimetic ubiquitin in mammalian cells. Molecular modeling based on the site-specific photo-crosslinking interaction map combined with mass spectrometry strongly suggests that a novel binding mechanism between Parkin and ubiquitin leads to a Parkin conformational change with subsequent activation of Parkin E3 ligase activity. PMID:26260794

  2. Tricyclic Covalent Inhibitors Selectively Target Jak3 through an Active Site Thiol*

    PubMed Central

    Goedken, Eric R.; Argiriadi, Maria A.; Banach, David L.; Fiamengo, Bryan A.; Foley, Sage E.; Frank, Kristine E.; George, Jonathan S.; Harris, Christopher M.; Hobson, Adrian D.; Ihle, David C.; Marcotte, Douglas; Merta, Philip J.; Michalak, Mark E.; Murdock, Sara E.; Tomlinson, Medha J.; Voss, Jeffrey W.

    2015-01-01

    The action of Janus kinases (JAKs) is required for multiple cytokine signaling pathways, and as such, JAK inhibitors hold promise for treatment of autoimmune disorders, including rheumatoid arthritis, inflammatory bowel disease, and psoriasis. However, due to high similarity in the active sites of the four members (Jak1, Jak2, Jak3, and Tyk2), developing selective inhibitors within this family is challenging. We have designed and characterized substituted, tricyclic Jak3 inhibitors that selectively avoid inhibition of the other JAKs. This is accomplished through a covalent interaction between an inhibitor containing a terminal electrophile and an active site cysteine (Cys-909). We found that these ATP competitive compounds are irreversible inhibitors of Jak3 enzyme activity in vitro. They possess high selectivity against other kinases and can potently (IC50 < 100 nm) inhibit Jak3 activity in cell-based assays. These results suggest irreversible inhibitors of this class may be useful selective agents, both as tools to probe Jak3 biology and potentially as therapies for autoimmune diseases. PMID:25552479

  3. Tricyclic Covalent Inhibitors Selectively Target Jak3 through an Active Site Thiol

    DOE PAGES

    Goedken, Eric R.; Argiriadi, Maria A.; Banach, David L.; ...

    2014-12-31

    The action of Janus kinases (JAKs) is required for multiple cytokine signaling pathways, and as such, JAK inhibitors hold promise for treatment of autoimmune disorders, including rheumatoid arthritis, inflammatory bowel disease, and psoriasis. However, due to high similarity in the active sites of the four members (Jak1, Jak2, Jak3, and Tyk2), developing selective inhibitors within this family is challenging. In this paper, we have designed and characterized substituted, tricyclic Jak3 inhibitors that selectively avoid inhibition of the other JAKs. This is accomplished through a covalent interaction between an inhibitor containing a terminal electrophile and an active site cysteine (Cys-909). Wemore » found that these ATP competitive compounds are irreversible inhibitors of Jak3 enzyme activity in vitro. They possess high selectivity against other kinases and can potently (IC 50 < 100 nM) inhibit Jak3 activity in cell-based assays. Finally, these results suggest irreversible inhibitors of this class may be useful selective agents, both as tools to probe Jak3 biology and potentially as therapies for autoimmune diseases.« less

  4. Tricyclic Covalent Inhibitors Selectively Target Jak3 through an Active Site Thiol

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Goedken, Eric R.; Argiriadi, Maria A.; Banach, David L.

    The action of Janus kinases (JAKs) is required for multiple cytokine signaling pathways, and as such, JAK inhibitors hold promise for treatment of autoimmune disorders, including rheumatoid arthritis, inflammatory bowel disease, and psoriasis. However, due to high similarity in the active sites of the four members (Jak1, Jak2, Jak3, and Tyk2), developing selective inhibitors within this family is challenging. In this paper, we have designed and characterized substituted, tricyclic Jak3 inhibitors that selectively avoid inhibition of the other JAKs. This is accomplished through a covalent interaction between an inhibitor containing a terminal electrophile and an active site cysteine (Cys-909). Wemore » found that these ATP competitive compounds are irreversible inhibitors of Jak3 enzyme activity in vitro. They possess high selectivity against other kinases and can potently (IC 50 < 100 nM) inhibit Jak3 activity in cell-based assays. Finally, these results suggest irreversible inhibitors of this class may be useful selective agents, both as tools to probe Jak3 biology and potentially as therapies for autoimmune diseases.« less

  5. Novel Autophosphorylation Sites of Src Family Kinases Regulate Kinase Activity and SH2 Domain Binding Capacity

    PubMed Central

    Weir, Marion E.; Mann, Jacqueline E.; Corwin, Thomas; Fulton, Zachary W.; Hao, Jennifer M.; Maniscalco, Jeanine F.; Kenney, Marie C.; Roque, Kristal M. Roman; Chapdelaine, Elizabeth F.; Stelzl, Ulrich; Deming, Paula B.; Ballif, Bryan A.; Hinkle, Karen L.

    2016-01-01

    Src family tyrosine kinases (SFKs) are critical players in normal and aberrant biological processes. While phosphorylation importantly-regulates SFKs at two known tyrosines, large-scale phosphoproteomics have revealed four additional tyrosines commonly-phosphorylated in SFKs. We found these novel tyrosines to be autophosphorylation sites. Mimicking phosphorylation at the site C-terminal to the activation loop decreased Fyn activity. Phosphomimetics and direct phosphorylation at the three SH2 domain sites increased Fyn activity while reducing phosphotyrosine-dependent interactions. While 68% of human SH2 domains exhibit conservation of at least one of these tyrosines, few have been found phosphorylated except when found in cis to a kinase domain. PMID:27001024

  6. Comparative Evaluation of Light-Trap Catches, Electric Motor Mosquito Catches and Human Biting Catches of Anopheles in the Three Gorges Reservoir

    PubMed Central

    Duo-quan, Wang; Lin-hua, Tang; Zhen-cheng, Gu; Xiang, Zheng; Man-ni, Yang; Wei-kang, Jiang

    2012-01-01

    The mosquito sampling efficiency of light-trap catches and electric motor mosquito catches were compared with that of human biting catches in the Three Gorges Reservoir. There was consistency in the sampling efficiency between light-trap catches and human biting catches for Anopheles sinensis (r = 0.82, P<0.01) and light-trap catches were 1.52 (1.35–1.71) times that of human biting catches regardless of mosquito density (r = 0.33, P>0.01), while the correlation between electric motor mosquito catches and human biting catches was found to be not statistically significant (r = 0.43, P>0.01) and its sampling efficiency was below that of human biting catches. It is concluded that light-traps can be used as an alternative to human biting catches of Anopheles sinensis in the study area and is a promising tool for sampling malaria vector populations. PMID:22235256

  7. Crystallographic Analysis of Active Site Contributions to Regiospecificity in the Diiron Enzyme Toluene 4-Monooxygenase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bailey, Lucas J.; Acheson, Justin F.; McCoy, Jason G.

    Crystal structures of toluene 4-monooxygenase hydroxylase in complex with reaction products and effector protein reveal active site interactions leading to regiospecificity. Complexes with phenolic products yield an asymmetric {mu}-phenoxo-bridged diiron center and a shift of diiron ligand E231 into a hydrogen bonding position with conserved T201. In contrast, complexes with inhibitors p-NH{sub 2}-benzoate and p-Br-benzoate showed a {mu}-1,1 coordination of carboxylate oxygen between the iron atoms and only a partial shift in the position of E231. Among active site residues, F176 trapped the aromatic ring of products against a surface of the active site cavity formed by G103, E104 andmore » A107, while F196 positioned the aromatic ring against this surface via a {pi}-stacking interaction. The proximity of G103 and F176 to the para substituent of the substrate aromatic ring and the structure of G103L T4moHD suggest how changes in regiospecificity arise from mutations at G103. Although effector protein binding produced significant shifts in the positions of residues along the outer portion of the active site (T201, N202, and Q228) and in some iron ligands (E231 and E197), surprisingly minor shifts (<1 {angstrom}) were produced in F176, F196, and other interior residues of the active site. Likewise, products bound to the diiron center in either the presence or absence of effector protein did not significantly shift the position of the interior residues, suggesting that positioning of the cognate substrates will not be strongly influenced by effector protein binding. Thus, changes in product distributions in the absence of the effector protein are proposed to arise from differences in rates of chemical steps of the reaction relative to motion of substrates within the active site channel of the uncomplexed, less efficient enzyme, while structural changes in diiron ligand geometry associated with cycling between diferrous and diferric states are discussed for their

  8. Evidence from molecular dynamics simulations of conformational preorganization in the ribonuclease H active site

    PubMed Central

    Stafford, Kate A.; Palmer III, Arthur G.

    2014-01-01

    Ribonuclease H1 (RNase H) enzymes are well-conserved endonucleases that are present in all domains of life and are particularly important in the life cycle of retroviruses as domains within reverse transcriptase. Despite extensive study, especially of the E. coli homolog, the interaction of the highly negatively charged active site with catalytically required magnesium ions remains poorly understood. In this work, we describe molecular dynamics simulations of the E. coli homolog in complex with magnesium ions, as well as simulations of other homologs in their apo states. Collectively, these results suggest that the active site is highly rigid in the apo state of all homologs studied and is conformationally preorganized to favor the binding of a magnesium ion. Notably, representatives of bacterial, eukaryotic, and retroviral RNases H all exhibit similar active-site rigidity, suggesting that this dynamic feature is only subtly modulated by amino acid sequence and is primarily imposed by the distinctive RNase H protein fold. PMID:25075292

  9. Active site structure and catalytic mechanism of phosphodiesterase for degradation of intracellular second messengers

    NASA Astrophysics Data System (ADS)

    Zhan, Chang-Guo

    2002-03-01

    Phosphodiesterases are clinical targets for a variety of biological disorders, because this superfamily of enzymes regulate intracellular concentration of cyclic nucleotides that serve as the second messengers playing a critical role in a variety of physiological processes. Understanding structure and mechanism of a phosphodiesterase will provide a solid basis for rational design of the more efficient therapeutics. Although a three-dimensional X-ray crystal structure of the catalytic domain of human phosphodiesterase 4B2B was recently reported, it was uncertain whether a critical bridging ligand in the active site is a water molecule or a hydroxide ion. The identity of this bridging ligand has been determined by performing first-principles quantum chemical calculations on models of the active site. All the results obtained indicate that this critical bridging ligand in the active site of the reported X-ray crystal structure is a hydroxide ion, rather than a water molecule, expected to serve as the nucleophile to initialize the catalytic degradation of the intracellular second messengers.

  10. Mutational Analysis of Escherichia coli MoeA: Two Functional Activities Map to the Active Site Cleft

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nichols,J.; Xiang, S.; Schindelin, H.

    2007-01-01

    The molybdenum cofactor is ubiquitous in nature, and the pathway for Moco biosynthesis is conserved in all three domains of life. Recent work has helped to illuminate one of the most enigmatic steps in Moco biosynthesis, ligation of metal to molybdopterin (the organic component of the cofactor) to form the active cofactor. In Escherichia coli, the MoeA protein mediates ligation of Mo to molybdopterin while the MogA protein enhances this process in an ATP-dependent manner. The X-ray crystal structures for both proteins have been previously described as well as two essential MogA residues, Asp49 and Asp82. Here we describe amore » detailed mutational analysis of the MoeA protein. Variants of conserved residues at the putative active site of MoeA were analyzed for a loss of function in two different, previously described assays, one employing moeA{sup -} crude extracts and the other utilizing a defined system. Oddly, no correlation was observed between the activity in the two assays. In fact, our results showed a general trend toward an inverse relationship between the activity in each assay. Moco binding studies indicated a strong correlation between a variant's ability to bind Moco and its activity in the purified component assay. Crystal structures of the functionally characterized MoeA variants revealed no major structural changes, indicating that the functional differences observed are not due to disruption of the protein structure. On the basis of these results, two different functional areas were assigned to regions at or near the MoeA active site cleft.« less

  11. Occurrence and source apportionment of polycyclic aromatic hydrocarbons in soils and sediment from Hanfeng Lake, Three Gorges, China.

    PubMed

    Cai, Jing; Gao, Shutao; Zhu, Like; Jia, Xuwei; Zeng, Xiangying; Yu, Zhiqiang

    2017-11-10

    This study was conducted to investigate the pollutant status and the retention mechanism of polycyclic aromatic hydrocarbons (PAHs) in soils and sediment from bank-water-level-fluctuating zone (WLFZ)-water systems in Hanfeng Lake, Three Gorges, China. The concentrations of the 16 PAHs ranged from 21.8 to 1324 ng g -1 dry wt for all 20 soil and sediment samples. These concentration levels were remarkably lower than those in soils and sediment collected domestically and worldwide. PAHs with two and three rings were found to be dominant in all the samples, with phenanthrene being most abundant. The spatial distribution of PAHs in bank soil, WLFZ soil, and sediment implied that the transfer and fate of PAHs in the bank soil-WLFZ soil-sediment systems were influenced by both water dynamic factors and physicochemical properties of PAHs. Diagnostic ratio analysis and principal component analysis suggested that the PAHs in the areas of Hanfeng Lake were primarily (>75%) derived from coal combustion and vehicle emissions . Use of natural gas, improving gasoline/diesel quality and phasing out old and nonstandard vehicles and ships are proposed to control PAH contamination and protect drinking water safety in the region.

  12. Assessing the potential for change in the middle Yangtze River channel following impoundment of the Three Gorges Dam

    NASA Astrophysics Data System (ADS)

    Yuan, Wenhao; Yin, Daowei; Finlayson, Brian; Chen, Zhongyuan

    2012-04-01

    The geomorphic impacts of dams on downstream river channels are complex, not readily predictable for specific cases, but widely reported in the literature. For the Three Gorges Dam on the Yangtze (Changjiang) River in China, no studies of the impact of the changed flow and sediment conditions below the dam on the behaviour of the channel were included in the pre-dam feasibility report. We have assembled a database of flow and sediment data for the middle Yangtze River from Yichang to Hankou and used this to analyse changes following the closure of the dam. While total flow is little affected, the operating strategy for the dam that provides for storage of part of the summer high flows to maintain hydroelectric power generation in winter (the low flow season) is reflected in changes to the seasonal distribution of flow below the dam. We calculated potential sediment carrying capacity and compared it with measured sediment concentrations for both pre- and post-dam conditions. While channel sedimentation is indicated along the middle Yangtze for pre-dam conditions, scour is indicated for post-dam conditions, highest at Yichang immediately below the dam and decreasing downstream.

  13. Silver-coated nylon dressing plus active DC microcurrent for healing of autogenous skin donor sites.

    PubMed

    Malin, Edward W; Galin, Chaya M; Lairet, Kimberley F; Huzar, Todd F; Williams, James F; Renz, Evan M; Wolf, Steven E; Cancio, Leopoldo C

    2013-11-01

    Burn wounds are a significant cause of morbidity and mortality, and improved outcomes are demonstrated with early closure of both primary burn wounds and skin donor sites. Thus, technology that decreases the healing time of burns and donor sites would be potentially lifesaving. We present the results of a single-center, prospective, double-blinded, randomized controlled trial to evaluate the efficacy of silver-coated dressing with active microcurrent in comparison to silver-coated dressing with sham microcurrent on wound-closure time for autogenous skin donor sites. Four hundred five patients were screened for treatment of their donor sites using a silver-coated nylon dressing with either sham or active microcurrent stimulation. Thirty patients were enrolled in the study and then randomized. Of these, 5 patients were removed from analysis due to protocol deviations. Differences in time-to-closure were analyzed using Kaplan-Meier analysis and the proportional hazard regression model. Subjective verbal pain rating scores (0-10; 0, no pain; 10, worst pain) were also recorded. All devices were blinded and programmed at an outside facility, so that every patient had either an active or sham device. The study was unblinded only after the final patient's donor site had healed. All patients achieved donor-site healing before postoperative day 20. The 14 patients in the active microcurrent group [mean, 10.8 (2.9) days; range, 7-15 days] experienced no difference in time to wound healing as compared to the remaining patients in the sham microcurrent group [mean, 11.1 (2.0) days; range, 8-14 days; P = 0.75]. There were no differences in pain from one group compared to the other. None of the donor sites exhibited clinical signs of infection. In a sample size of 25 burn patients, the addition of direct microcurrent to silver-nylon dressings did not decrease time to wound closure of skin donor sites, and it did not show a difference in reported pain levels.

  14. STRUCTURAL AND FUNCTIONAL CONSEQUENCES OF CIRCULAR PERMUTATION ON THE ACTIVE SITE OF OLD YELLOW ENZYME.

    PubMed

    Daugherty, Ashley B; Horton, John R; Cheng, Xiaodong; Lutz, Stefan

    2015-02-06

    Circular permutation of the NADPH-dependent oxidoreductase Old Yellow Enzyme from Saccharomyces pastorianus (OYE1) can significantly enhance the enzyme's catalytic performance. Termini relocation into four regions of the protein (sectors I-IV) near the active site has proven effective in altering enzyme function. To better understand the structural consequences and rationalize the observed functional gains in these OYE1 variants, we selected representatives from sectors I-III for further characterization by biophysical methods and X-ray crystallography. These investigations not only show trends in enzyme stability and quaternary structure as a function of termini location, but also provide a possible explanation for the catalytic gains in our top-performing OYE variant (new N-terminus at residue 303; sector III). Crystallographic analysis indicates that termini relocation into sector III affects the loop β6 region (amino acid positions: 290-310) of OYE1 which forms a lid over the active site. Peptide backbone cleavage greatly enhances local flexibility, effectively converting the loop into a tether and consequently increasing the environmental exposure of the active site. Interestingly, such active site remodeling does not negatively impact the enzyme's activity and stereoselectivity, nor does it perturb the conformation of other key active site residues with the exception of Y375. These observations were confirmed in truncation experiments, deleting all residues of the loop β6 region in our OYE variant. Intrigued by the finding that circular permutation leaves most of the key catalytic residues unchanged, we also tested OYE permutants for possible additive or synergistic effects of amino acid substitutions. Distinct functional changes in these OYE variants were detected upon mutations at W116, known in native OYE1 to cause inversion of diastereo-selectivity for ( S )-carvone reduction. Our findings demonstrate the contribution of loop β6 toward determining the

  15. Non-canonical active site architecture of the radical SAM thiamin pyrimidine synthase

    DOE PAGES

    Fenwick, Michael K.; Mehta, Angad P.; Zhang, Yang; ...

    2015-03-27

    Radical S-adenosylmethionine (SAM) enzymes use a [4Fe-4S] cluster to generate a 5'-deoxyadenosyl radical. Canonical radical SAM enzymes are characterized by a β-barrel-like fold and SAM anchors to the differentiated iron of the cluster, which is located near the amino terminus and within the β-barrel, through its amino and carboxylate groups. Here we show that ThiC, the thiamin pyrimidine synthase in plants and bacteria, contains a tethered cluster-binding domain at its carboxy terminus that moves in and out of the active site during catalysis. In contrast to canonical radical SAM enzymes, we predict that SAM anchors to an additional active sitemore » metal through its amino and carboxylate groups. Superimposition of the catalytic domains of ThiC and glutamate mutase shows that these two enzymes share similar active site architectures, thus providing strong evidence for an evolutionary link between the radical SAM and adenosylcobalamin-dependent enzyme superfamilies.« less

  16. Active site-directed double mutants of dihydrofolate reductase.

    PubMed

    Ercikan-Abali, E A; Mineishi, S; Tong, Y; Nakahara, S; Waltham, M C; Banerjee, D; Chen, W; Sadelain, M; Bertino, J R

    1996-09-15

    Variants of dihydrofolate reductase (DHFR), which confer resistance to antifolates, are used as dominant selectable markers in vitro and in vivo and may be useful in the context of gene therapy. To identify improved mutant human DHFRs with increased catalytic efficiency and decreased binding to methotrexate, we constructed by site-directed mutagenesis four variants with substitutions at both Leu22 and Phe31 (i.e., Phe22-Ser31, Tyr22-Ser31, Phe22-Gly31, and Tyr22-Gly31). Antifolate resistance has been observed previously when individual changes are made at these active-site residues. Substrate and antifolate binding properties of these "double" mutants revealed that each have greatly diminished affinity for antifolates (> 10,000-fold) yet only slightly reduced substrate affinity. Comparison of in vitro measured properties with those of single-residue variants indicates that double mutants are indeed significantly superior. This was verified for one of the double mutants that provided high-level methotrexate resistance following retrovirus-mediated gene transfer in NIH3T3 cells.

  17. Impact of the Three-Gorges Dam and water transfer project on Changjiang floods

    NASA Astrophysics Data System (ADS)

    Nakayama, Tadanobu; Shankman, David

    2013-01-01

    Increasing frequency of severe floods on the middle and lower Changjiang (Yangtze) River during the past few decades can be attributed to both abnormal monsoon rainfall and landscape changes that include extensive deforestation affecting river sedimentation, and shrinking lakes and levee construction that reduced the areas available for floodwater storage. The Three-Gorges Dam (TGD) and the South-to-North Water Transfer Project (SNWTP) will also affect frequency and intensity of severe floods in the Poyang Lake region of the middle Changjiang. Process-based National Integrated Catchment-based Eco-hydrology (NICE) model predicts that the TGD will increase flood risk during the early summer monsoon against the original justifications for building the dam, relating to complex river-lake-groundwater interactions. Several scenarios predict that morphological change will increase flood risk around the lake. This indicates the importance of managing both flood discharge and sediment deposition for the entire basin. Further, the authors assessed the impact of sand mining in the lake after its prohibition on the Changjiang, and clarified that alternative scenario of sand mining in lakes currently disconnected from the mainstream would reduce the flood risk to a greater extent than intensive dredging along junction channel. Because dry biomasses simulated by the model were linearly related to the Time-Integrated Normalized Difference Vegetation Index (TINDVI) estimated from satellite images, its decadal gradient during 1982-1999 showed a spatially heterogeneous distribution and generally decreasing trends beside the lakes, indicating that the increases in lake reclamation and the resultant decrease in rice productivity are closely related to the hydrologic changes. This integrated approach could help to minimize flood damage and promote better decisions addressing sustainable development.

  18. Stereoselectivity of supported alkene metathesis catalysts: a goal and a tool to characterize active sites.

    PubMed

    Copéret, Christophe

    2011-01-05

    Stereoselectivity in alkene metathesis is a challenge and can be used as a tool to study active sites under working conditions. This review describes the stereochemical relevance and problems in alkene metathesis (kinetic vs. thermodynamic issues), the use of (E/Z) ratio at low conversions as a tool to characterize active sites of heterogeneous catalysts and finally to propose strategies to improve catalysts based on the current state of the art.

  19. Interaction of mining activities and aquatic environment: A review from Greek mine sites.

    NASA Astrophysics Data System (ADS)

    Vasileiou, Eleni; Kallioras, Andreas

    2016-04-01

    In Greece a significant amount of mineral and ore deposits have been recorded accompanied by large industrial interest and a long mining history. Today many active and/or abandoned mine sites are scattered within the country; while mining activities take place in different sites for exploiting various deposits (clay, limestone, slate, gypsum, kaolin, mixed sulphide ores (lead, zinc, olivine, pozzolan, quartz lignite, nickel, magnesite, aluminum, bauxite, gold, marbles etc). The most prominent recent ones are: (i) the lignite exploitation that is extended in the area of Ptolemais (Western Macedonia) and Megalopolis (Central Peloponnese); and (ii) the major bauxite deposits located in central Greece within the Parnassos-Ghiona geotectonic zone and on Euboea Island. In the latter area, significant ores of magnesite were exploited and mixed sulphide ores. Centuries of intensive mining exploitation and metallurgical treatment of lead-silver deposits in Greece, have also resulted in significant abandoned sites, such as the one in Lavrion. Mining activities in Lavrio, were initiated in ancient times and continued until the 1980s, resulting in the production of significant waste stockpiles deposited in the area, crucial for the local water resources. Ιn many mining sites, environmental pressures are also recorded after the mine closure to the aquatic environment, as the surface waters flow through waste dump areas and contaminated soils. This paper aims to the geospatial visualization of the mining activities in Greece, in connection to their negative (surface- and/or ground-water pollution; overpumping due to extensive dewatering practices) or positive (enhanced groundwater recharge; pit lakes, improvement of water budget in the catchment scale) impacts on local water resources.

  20. Resonance energy transfer between the active sites of creatine kinase from rabbit brain.

    PubMed

    Grossman, S H

    1990-09-03

    Resonance energy transfer was measured between the active site domains of the brain isozyme of creatine kinase (CK-BB). The reactive thiol near the active sites, one on each subunit of the dimeric protein, was derivatized using 5-[2-[iodoacetyl)amino)ethyl]aminonaphthalene-1-sulfonic acid (AED), 2-[4'-iodoacetamidoanilino]naphthalene-6-sulfonic acid (AANS) and 5-iodoacetamidofluorescein (AF). Suitable donor/acceptor protein conjugated hybrids were prepared by controlled kinetics producing CK-BB-AED/AF and CK-BB-AANS/AF. Transfer efficiencies, measured from the quenching of the donor lifetime and steady-state sensitized acceptor emission, ranged from 0.10 to 0.17. From determination of the donor/acceptor overlap integrals, donor quantum yields and attempts to delimit the orientation factor using steady-state and phase-resolved anisotropy measurements, it was found that a suitable estimate of the range between the active sites was between 45 and 57 A. This range is similar to that reported previously for the muscle isozyme of creatine kinase (Grossman, S.H. (1989) Biochemistry 28, 4894-4902) but is a significantly greater distance than detected for the hybrid, myocardial specific isozyme (Grossman, S.H. (1983) Biochemistry 22, 5369-5375).