Impacts of global warming on phenology of spring leaf unfolding remain stable in the long run.

PubMed

Wang, Huanjiong; Rutishauser, This; Tao, Zexing; Zhong, Shuying; Ge, Quansheng; Dai, Junhu

2017-02-01

The impact of spring temperature forcing on the timing of leaf unfolding of plants (temperature sensitivity, S T ) is one important indicator of how and to what degree plant species track climate change. Fu et al. (Nature 526:104-107, 2015) found that S T has significantly decreased from the 1980-1994 to the 1999-2013 period for seven mid-latitude tree species in Europe. However, long-term changes in S T over the past 60 years are still not clear. Here, using in situ observations of leaf unfolding for seven dominant European tree species, we analyze the temporal change in S T over decadal time scales extending the data series back to 1951. Our results demonstrate that S T shows no statistically significant change within shifting 30-year windows from 1951 to 2013 and remains stable between 1951-1980 and 1984-2013 (3.6 versus 3.7 days °C -1 ). This result suggests that the significant decrease in S T over the past 33 years could not be sustained when examining the trends of phenological responses in the long run. Therefore, we could not conclude that tree spring phenology advances will slow down in the future, and the S T changes in warming scenarios are still uncertain.

Phenology, Canopy Aging and Seasonal Carbon Balance as Related to Delayed Winter Pruning of Vitis vinifera L. cv. Sangiovese Grapevines

PubMed Central

Gatti, Matteo; Pirez, Facundo J.; Chiari, Giorgio; Tombesi, Sergio; Palliotti, Alberto; Merli, Maria C.; Poni, Stefano

2016-01-01

Manipulating or shifting annual grapevine growing cycle to offset limitations imposed by global warming is a must today, and delayed winter pruning is a tool to achieve it. However, no information is available about its physiological background, especially in relation to modifications in canopy phenology, demography and seasonal carbon budget. Mechanistic hypothesis underlying this work was that very late winter pruning (LWP) can achieve significant postponement of phenological stages so that ripening might occur in a cooler period and, concurrently, ripening potential can be improved due to higher efficiency and prolonged longevity of the canopy. Variability in the dynamics of the annual cycle was created in mature potted cv. Sangiovese grapevines subjected to either standard winter pruning (SWP) or late and very late winter pruning (LWP, VLWP) performed when apical shoots on the unpruned canes were at the stage of 2 and 7 unfolded leaves. Vegetative growth, phenology and canopy net CO2 exchange (NCER) were followed throughout the season. Despite LWP and VLWP induced a bud-burst delay of 17 and 31 days vs. SWP, the delay was fully offset at harvest for LWP and was reduced to 6 days in VLWP. LWP showed notably higher canopy efficiency as shorter time needed to reach maximum NCER/leaf area (22 days vs. 34 in SWP), highest maximum NCER/leaf area (+37% as compared to SWP) and higher NCER/leaf area rates from veraison to end of season. As a result, seasonal cumulated carbon in LWP was 17% higher than SWP. A negative functional relationship was also established between amount of leaf area removed at winter pruning and yield per vine and berry number per cluster. Although retarded winter pruning was not able to postpone late-season phenological stages under the warm conditions of this study, it showed a remarkable potential to limit yield while improving grape quality, thereby fostering the hypothesis that it could be used to replace time-consuming and costly cluster thinning. This preliminary study indicates that proper winter pruning date should be timed so as not to exceed the stage of two unfolded leaves. PMID:27242860

Effects of extreme spring temperatures on phenology: a case study from Munich and Ingolstadt

NASA Astrophysics Data System (ADS)

Jochner, Susanne; Menzel, Annette

2010-05-01

Extreme events - e.g. warm spells or heavy precipitation events - are likely to increase in the future both in frequency and intensity. Therefore, research on extreme events gains new importance; also in terms of plant development which is mostly triggered by temperatures. An arising question is how plants respond to an extreme warm spell when following an extreme cold winter season. This situation could be studied in spring 2009 in the greater area of Munich and Ingolstadt by phenological observations of flowering and leaf unfolding of birch (Betula pendula L.) and flowering of horse chestnut (Aesculus hippocastanum L.). The long chilling period of winter 2008 and spring 2009 was followed by an immediate strong forcing of flowering and leaf unfolding, especially for birch. This extreme weather situation diminished the difference between urban and rural dates of onset. Another important fact that could be observed in the proceeding period of December 2008 to April 2009 was the reduced temperature difference among urban and rural sites (urban heat island effect). Long-term observations (1951-2008) of the phenological network of the German Meteorological Service (DWD) were used to identify years with reduced urban-rural differences between onset times in the greater area of Munich in the past. Statistical analyses were conducted in order to answer the question whether the sequence of extreme warm and cold events leads to a decreased difference in phenological onset times or if this behaviour can be attributed to extreme warm springs themselves or to the decreased urban heat island effect which is mostly affected by general atmospheric circulation patterns.

Trend of earlier spring in central Europe continued

NASA Astrophysics Data System (ADS)

Ungersböck, Markus; Jurkovic, Anita; Koch, Elisabeth; Lipa, Wolfgang; Scheifinger, Helfried; Zach-Hermann, Susanne

2013-04-01

Modern phenology is the study of the timing of recurring biological events in the animal and plant world, the causes of their timing with regard to biotic and abiotic forces, and the interrelation among phases of the same or different species. The relationship between phenology and climate explains the importance of plant phenology for Climate Change studies. Plants require light, water, oxygen mineral nutrients and suitable temperature to grow. In temperate zones the seasonal life cycle of plants is primarily controlled by temperature and day length. Higher spring air temperatures are resulting in an earlier onset of the phenological spring in temperate and cool climate. On the other hand changes in phenology due to climate change do have impact on the climate system itself. Vegetation is a dynamic factor in the earth - climate system and has positive and negative feedback mechanisms to the biogeochemical and biogeophysical fluxes to the atmosphere Since the mid of the 1980s spring springs earlier in Europe and autumn is shifting back to the end of the year resulting in a longer vegetation period. The advancement of spring can be clearly attributed to temperature increase in the months prior to leaf unfolding and flowering, the timing of autumn is more complex and cannot easily be attributed to one or some few parameters. To demonstrate that the observed advancement of spring since the mid of 1980s is pro-longed in 2001 to 2010 and the delay of fall and the lengthening of the growing season is confirmed in the last decade we picked out several indicator plants from the PEP725 database www.pep725.eu. The PEP725 database collects data from different European network operators and thus offers a unique compilation of phenological observations; the database is regularly updated. The data follow the same classification scheme, the so called BBCH coding system so they can be compared. Lilac Syringa vulgaris, birch Betula pendula, beech Fagus and horse chestnut Aesculus hippocastanum are well represented in the PEP725 database. Flowering of lilac Syringa vulgaris is also used in the US as spring indicator . The flowering and/or leaf unfolding dates of lilac, horse chestnut show a clear advance to an earlier entrance in the last two decades 1991 to 2000 and 2001 to 2010 compared with the reference period 1961 to 1990, being more pronounced in northwestern regions of Central Europe. The growing season defined here as time span between leaf unfolding and leaf coloration of birch and beech has been lengthening up to two weeks in 2001 to 2010 compared to 1961 to 1990 in northeastern parts of Central Europe.

Change in First Leaf Date Between 1951-1960 and 2006-2015

EPA Pesticide Factsheets

This figure shows modeled trends in lilac and honeysuckle first leaf dates at weather stations across the contiguous 48 states. This map compares the average first leaf date during two 10-year periods, developed using data from the USA National Phenology Network. Blue circles represent later leaf dates, and red circles represent earlier. For more information: www.epa.gov/climatechange/science/indicators

Phenological behaviour of early spring flowering trees in Spain in response to recent climate changes

NASA Astrophysics Data System (ADS)

Hidalgo-Galvez, M. D.; García-Mozo, H.; Oteros, J.; Mestre, A.; Botey, R.; Galán, C.

2018-04-01

This research reports the phenological trends of four early spring and late winter flowering trees in Spain (south Europe) from a recent period (1986-2012). The studied species were deciduous trees growing in different climatic areas: hazel ( Corylus avellana L.), willow ( Salix alba L.), ash ( Fraxinus angustifolia Vahl.) and white mulberry ( Morus alba L.). We analysed the response to climate and the trends of the following phenophases observed at the field: budburst, leaf unfolding, flowering, fruit ripening, fruit harvesting, leaf colour change and leaf-fall. The study was carried out in 17 sampling sites in the country with the aim of detecting the recent phenological response to the climate of these species, and the possible effect of climate change. We have observed differences in the phenological response to climate depending on each species. Sixty-one percent of studied sites suffered an advance of early spring phenophases, especially budburst on average by -0.67 days and flowering on average by -0.15 days during the studied period, and also in the subsequent fruit ripening and harvesting phases on average by -1.06 days. By contrast, it has been detected that 63% of sampling sites showed a delay in autumn vegetative phases, especially leaf-fall events on average by +1.15 days. The statistic correlation analysis shows in the 55% of the studied localities that phenological advances are the consequence of the increasing trend detected for temperature—being minimum temperature the most influential factor—and in the 52% of them, phenological advances occurred by rainfall variations. In general, leaf unfolding and flowering from these species showed negative correlations in relation to temperature and rainfall, whereas that leaf colour change and leaf-fall presented positive correlations. The results obtained have a great relevance due to the fact that they can be considered as reliable bio-indicators of the impact of the recent climate changes in southern Europe.

Day length unlikely to constrain climate-driven shifts in leaf-out times of northern woody plants

NASA Astrophysics Data System (ADS)

Zohner, Constantin M.; Benito, Blas M.; Svenning, Jens-Christian; Renner, Susanne S.

2016-12-01

The relative roles of temperature and day length in driving spring leaf unfolding are known for few species, limiting our ability to predict phenology under climate warming. Using experimental data, we assess the importance of photoperiod as a leaf-out regulator in 173 woody species from throughout the Northern Hemisphere, and we also infer the influence of winter duration, temperature seasonality, and inter-annual temperature variability. We combine results from climate- and light-controlled chambers with species’ native climate niches inferred from georeferenced occurrences and range maps. Of the 173 species, only 35% relied on spring photoperiod as a leaf-out signal. Contrary to previous suggestions, these species come from lower latitudes, whereas species from high latitudes with long winters leafed out independent of photoperiod. The strong effect of species’ geographic-climatic history on phenological strategies complicates the prediction of community-wide phenological change.

Quantitative inheritance of crop timing traits in interspecific hybrid Petunia populations and interactions with crop quality parameters.

PubMed

Warner, Ryan M; Walworth, Aaron E

2010-01-01

The leaf unfolding rate (i.e., development rate) and the number of nodes forming prior to floral initiation are 2 factors determining production times for floriculture crops. Wild relative species of the cultivated petunia (Petunia x hybrida Vilm.) that exhibited faster development rates than modern cultivars and may therefore be useful genetic sources to develop cultivars with decreased production time were identified. Three interspecific F(2) families, Petunia exserta Stehmann x P. axillaris (Lam.) Britton et al., P. x hybrida 'Mitchell' x P. axillaris, and P. axillaris x P. integrifolia (Hook.) Schinz & Thell. all exhibited transgressive segregation for development rate and node number below the first flower. Development rate and time to flower segregated independently in all families. Leaf number below the first flower was positively correlated with leaf unfolding rate in all families except P. axillaris x P. integrifolia. Time to flower was positively correlated with flower bud number in the P. x hybrida 'Mitchell' x P. axillaris and P. axillaris x P. integrifolia families only. Based on these results, wild Petunia germplasm should be useful for developing petunia cultivars with reduced crop production times, but some negative effects on crop quality parameters may need to be overcome.

Modelling of the spring phenological phases of the Silver birch Betula pendula and Bird cherry Padus racemosa in Baltic region

NASA Astrophysics Data System (ADS)

Kalvāns, Andis; Kalvāne, Gunta; Bitāne, Māra; Cepīte-Frišfelde, Daiga; Sīle, Tija; Seņņikovs, Juris

2014-05-01

The air temperature is the strongest driving factor of the plant development during spring time in moderate climate conditions. However other factors such as the air temperature during the dormancy period and light conditions can play a role as well. The full potential of the recent and historical phenological observation data can be utilised by modelling tools. We have calibrated seven phenological models described in scientific literature to calculate the likely dates leaf unfolding and start of flowering of the Silver birch Betula pendula and bird cherry Padus racemosa (Kalvāns at al, accepted). Phenological observations are derived from voluntary observation network for period 1960-2009 in Latvia. The number of used observations for each phase range from 149 to 172. Air temperature data measured in meteorological stations closest to the corresponding phenological observation sites are obtained from Latvian Environment, Geology and Meteorology Centre. We used 33 random data subsamples for model calibration to produce a range of model coefficients enabling the estimation of the phenological model uncertainty. It is found that the best reproduction of the observational data are obtained using a simple linear degree day model considering daily minimum and maximum temperature and more complex sigmoidal model honouring the need for low temperatures for dormancy release (UniChill, Chuine, 2000). The median calibration base temperature in the degree day model for the silver birch leaf unfolding is 5.6°C and for start of the flowering 6.7°C; for the bird cherry the corresponding base temperatures are 3.2°C and 3.4°C. The calibrated models and air temperature archive data derived from the Danish Meteorological Institute is used to simulate the respective phase onset in the Estonia, Latvia and Lithuania in 2009. Significant regional differences between modelled phase onset times are observed. There is a wide regional variation of the model uncertainty as well, indicated by the confidence intervals calculated from the 33 model calibrations: in some regions all the coefficient sets give similar phase onset times (within two to three day interval) while on other cases the spread can be more than a weak. In the spring 2014 field campaign is planned to obtain considerable data set for leaf unfolding and start of flowering of the bird cherry in Estonia, Latvia and Lithuania. The data will be used to evaluate performance of phenological models driven by short, medium and long term air temperature forecasts. The research is supported by the European Union through the European Social Fund Mobilitas grant No MJD309. References Chuine, I. (2000). A unified model for budburst of trees. Journal of theoretical biology, 207 (3), 337-347 Kalvāns, A., Bitāne, M., Kalvāne, G., accepted. Forecasting plant phenology: evaluating the phenological models for Betula pendula and Padus racemosa spring phases, Latvia. International Journal of Biometeorology, accepted.

Dating Violence, Bullying, and Sexual Harassment: Longitudinal Profiles and Transitions over Time

ERIC Educational Resources Information Center

Miller, Shari; Williams, Jason; Cutbush, Stacey; Gibbs, Deborah; Clinton-Sherrod, Monique; Jones, Sarah

2013-01-01

Although there is growing recognition of the problem of dating violence, little is known about how it unfolds among young adolescents who are just beginning to date. This study examined classes (subgroups) and transitions between classes over three time points based on dating violence, bullying, and sexual harassment perpetration and victimization…

Phylogenetic conservatism and trait correlates of spring phenological responses to climate change in northeast China.

PubMed

Du, Yanjun; Chen, Jingru; Willis, Charles G; Zhou, Zhiqiang; Liu, Tong; Dai, Wujun; Zhao, Yuan; Ma, Keping

2017-09-01

Climate change has resulted in major changes in plant phenology across the globe that includes leaf-out date and flowering time. The ability of species to respond to climate change, in part, depends on their response to climate as a phenological cue in general. Species that are not phenologically responsive may suffer in the face of continued climate change. Comparative studies of phenology have found phylogeny to be a reliable predictor of mean leaf-out date and flowering time at both the local and global scales. This is less true for flowering time response (i.e., the correlation between phenological timing and climate factors), while no study to date has explored whether the response of leaf-out date to climate factors exhibits phylogenetic signal. We used a 52-year observational phenological dataset for 52 woody species from the Forest Botanical Garden of Heilongjiang Province, China, to test phylogenetic signal in leaf-out date and flowering time, as well as, the response of these two phenological traits to both temperature and winter precipitation. Leaf-out date and flowering time were significantly responsive to temperature for most species, advancing, on average, 3.11 and 2.87 day/°C, respectively. Both leaf-out and flowering, and their responses to temperature exhibited significant phylogenetic signals. The response of leaf-out date to precipitation exhibited no phylogenetic signal, while flowering time response to precipitation did. Native species tended to have a weaker flowering response to temperature than non-native species. Earlier leaf-out species tended to have a greater response to winter precipitation. This study is the first to assess phylogenetic signal of leaf-out response to climate change, which suggests, that climate change has the potential to shape the plant communities, not only through flowering sensitivity, but also through leaf-out sensitivity.

Influences of the El Nino Southern Oscillation and the Pacific Decadal Oscillation on the Timing of the North American Spring

NASA Technical Reports Server (NTRS)

McCabe, Gregory J.; Ault, Toby R.; Cook, Benjamin I.; Betancourt, Julio L.; Schwartz, Mark D.

2012-01-01

Detrended, modelled first leaf dates for 856 sites across North America for the period 1900-2008 are used to examine how the El Nino Southern Oscillation (ENSO) and the Pacific Decadal Oscillation (PDO) separately and together might influence the timing of spring. Although spring (mean March through April) ENSO and PDO signals are apparent in first leaf dates, the signals are not statistically significant (at a 95% confidence level (p <0.05)) for most sites. The most significant ENSO/PDO signal in first leaf dates occurs for El Nino and positive PDO conditions. An analysis of the spatial distributions of first leaf dates for separate and combined ENSO/PDO conditions features a northwest-southeast dipole that is significantly (at p <0.05) different than the distributions for neutral conditions. The nature of the teleconnection between Pacific SST's and first leaf dates is evident in comparable composites for detrended sea level pressure (SLP) in the spring months. During positive ENSO/PDO, there is an anomalous flow of warm air from the southwestern US into the northwestern US and an anomalous northeasterly flow of cold air from polar regions into the eastern and southeastern US. These flow patterns are reversed during negative ENSO/PDO. Although the magnitudes of first leaf date departures are not necessarily significantly related to ENSO and PDO, the spatial patterns of departures are significantly related to ENSO and PDO. These significant relations and the long-lived persistence of SSTs provide a potential tool for forecasting the tendencies for first leaf dates to be early or late.

[Several changes of Indocalamus leaf active ingredients contents].

PubMed

Su, Chun-hua; Liu, Guo-hua; Wang, Fu-sheng; Ding, Yu-long; Xue, Jian-hui

2011-09-01

In this paper, the leaves of Indocalamus herklotsii, Indocalamus decorus, and Indocalamus latifolius were collected from Nanjing in different seasons to study the seasonal changes of the total flavonoids, tea polyphenols, and soluble sugar contents in the leaves. There existed significant differences in the test active ingredients contents among the leaves of the three Indocalamus species. The leaf total flavonoids content of the three Indocalamus species in different seasons ranged in 1.7%-2.7%, being the highest for I. herklotsii and I. decorus in spring and for I. latifolius in winter. The leaf tea polyphenols content varied from 5.5% to 7.6%; and the leaf soluble sugar content was 1.0%-8.5%, with the maximum in spring. Within the three months after leaf unfolding, the active ingredients contents in I. herklotsii and I. decorus leaves increased with leaf age. The optimal period for harvesting Indocalamus leaves was from December to next March. Among the three Indocalamus species, I. latifolius had the highest contents of the three active ingredients in leaves, suggesting that I. latifolius had greater potential value in the utilization of its leaf active ingredients than the other two species.

Determining past leaf-out times of New England's deciduous forests from herbarium specimens.

PubMed

Everill, Peter H; Primack, Richard B; Ellwood, Elizabeth R; Melaas, Eli K

2014-08-01

• There is great interest in studying leaf-out times of temperate forests because of the importance of leaf-out in controlling ecosystem processes, especially in the face of a changing climate. Remote sensing and modeling, combined with weather records and field observations, are increasing our knowledge of factors affecting variation in leaf-out times. Herbarium specimens represent a potential new source of information to determine whether the variation in leaf-out times observed in recent decades is comparable to longer time frames over past centuries.• Here we introduce the use of herbarium specimens as a method for studying long-term changes in leaf-out times of deciduous trees. We collected historical leaf-out data for the years 1834-2008 from common deciduous trees in New England using 1599 dated herbarium specimens with young leaves.• We found that leaf-out dates are strongly affected by spring temperature, with trees leafing out 2.70 d earlier for each degree C increase in mean April temperature. For each degree C increase in local temperature, trees leafed out 2.06 d earlier. Additionally, the mean response of leaf-out dates across all species and sites over time was 0.4 d earlier per decade. Our results are of comparable magnitude to results from studies using remote sensing and direct field observations.• Across New England, mean leaf-out dates varied geographically in close correspondence with those observed in studies using satellite data. This study demonstrates that herbarium specimens can be a valuable source of data on past leaf-out times of deciduous trees. © 2014 Botanical Society of America, Inc.

Influences of the El Niño Southern Oscillation and the Pacific Decadal Oscillation on the timing of the North American spring

USGS Publications Warehouse

McCabe, Gregory J.; Ault, Toby R.; Cook, Benjamin I.; Betancourt, Julio L.; Schwartz, Mark D.

2012-01-01

Detrended, modelled first leaf dates for 856 sites across North America for the period 1900–2008 are used to examine how the El Niño Southern Oscillation (ENSO) and the Pacific Decadal Oscillation (PDO) separately and together might influence the timing of spring. Although spring (mean March through April) ENSO and PDO signals are apparent in first leaf dates, the signals are not statistically significant (at a 95% confidence level (p < 0.05)) for most sites. The most significant ENSO/PDO signal in first leaf dates occurs for El Niño and positive PDO conditions. An analysis of the spatial distributions of first leaf dates for separate and combined ENSO/PDO conditions features a northwest–southeast dipole that is significantly (at p < 0.05) different than the distributions for neutral conditions. The nature of the teleconnection between Pacific SST's and first leaf dates is evident in comparable composites for detrended sea level pressure (SLP) in the spring months. During positive ENSO/PDO, there is an anomalous flow of warm air from the southwestern US into the northwestern US and an anomalous northeasterly flow of cold air from polar regions into the eastern and southeastern US. These flow patterns are reversed during negative ENSO/PDO. Although the magnitudes of first leaf date departures are not necessarily significantly related to ENSO and PDO, the spatial patterns of departures are significantly related to ENSO and PDO. These significant relations and the long-lived persistence of SSTs provide a potential tool for forecasting the tendencies for first leaf dates to be early or late.

PEP725: real time monitoring of phenological events in Austria, Germany, Sweden and Switzerland

NASA Astrophysics Data System (ADS)

Ungersboeck, Markus; Bolmgren, Kjell; Huebner, Thomas; Kaspar, Frank; Langvall, Ola; Paul, Anita; Pietragalla, Barbara; Scheifinger, Helfried; Koch, Elisabeth

2017-04-01

The main objective of PEP725 (Pan European Phenological database; http://www.pep725.eu/) is to promote and facilitate phenological research by delivering a pan European phenological database with an open, unrestricted data access for science, research and education. The first datasets in PEP725 date back to 1868; however, there are only a few observations available until 1950. From 1951 onwards, the phenological networks all over Europe developed rapidly. So far more than 11 923 489 of observations of 121 different plants are now available in the PEP725 database. Approximately 40 % of all data are flowering records, 10 % are fruit ripeness observations and also 10 % are leaf unfolding observations. The PEP725 database is updated annually. But since recently Deutscher Wetterdienst and MeteoSwiss offer their observers to upload their observations via web in real time mode, ZAMG introduced this web-based feature already in 2007 (phenowatch.at) and the observers of SWE-NPN (the Swedish National Phenology Network) can submit their observations through the web application naturenskalender.se since the start in 2008. Since spring 2016 one you can find a real time animated monitoring tool showing how the "green wave" in spring is moving from 46° northern latitude up to the Arctic Circle and the "brown wave" in autumn in the opposite direction. In 2015 the "green wave" speeds up from app. 4.4 days/degree latitude for hazel flowering to 2.9 days/ degree latitude for willow flowering and 2.25 days/degree latitude for birch leaf unfolding. There are other European countries as for instance Italy, The Netherlands, UK that have been doing visualizations of ground phenology in real time for some years, but these efforts always end at the national borders. PEP725 is funded by ZAMG, the Austrian ministry of science, research and economy and EUMETNET, the network of European meteorological services. So far 21 European meteorological services and 7 partners from different phenological network operators have joined PEP725.

A Global Regulation Inducing the Shape of Growing Folded Leaves

PubMed Central

Couturier, Etienne; Courrech du Pont, Sylvain; Douady, Stéphane

2009-01-01

Shape is one of the important characteristics for the structures observed in living organisms. Whereas biologists have proposed models where the shape is controlled on a molecular level [1], physicists, following Turing [2] and d'Arcy Thomson [3], have developed theories where patterns arise spontaneously [4]. Here, we propose that volume constraints restrict the possible shapes of leaves. Focusing on palmate leaves (with lobes), the central observation is that developing leaves first grow folded inside a bud, limited by the previous and subsequent leaves. We show that the lobe perimeters end at the border of this small volume. This induces a direct relationship between the way it was folded and the final unfolded shape of the leaf. These dependencies can be approximated as simple geometrical relationships that we confirm on both folded embryonic and unfolded mature leaves. We find that independent of their position in the phylogenetic tree, these relationships work for folded species, but do not work for non-folded species. This global regulation for the leaf growth could come from a mechanical steric constraint. Such steric regulation should be more general and considered as a new simple means of global regulation. PMID:19956690

Multidecadal Changes and Interannual Variation in Springtime Phenology of North American Temperate and Boreal Deciduous Forests

NASA Astrophysics Data System (ADS)

Melaas, Eli K.; Sulla-Menashe, Damien; Friedl, Mark A.

2018-03-01

The timing of leaf emergence is an important diagnostic of climate change impacts on ecosystems. Here we present the first continental-scale analysis of multidecadal changes in the timing of spring onset across North American temperate and boreal forests based on Landsat imagery. Our results show that leaf emergence in Eastern Temperate Forests has consistently trended earlier, with a median change of about 1 week over the 30 year study period. Changes in leaf emergence dates in boreal forests were more heterogeneous, with some sites showing trends toward later dates. Interannual variability in leaf emergence dates was strongly sensitive to springtime accumulated growing degree days across all sites, and geographic patterns of changes in onset dates were highly correlated with changes in regional springtime temperatures. These results provide a refined characterization of recent changes in springtime forest phenology and improve understanding regarding the sensitivity of North American forests to climate change.

Spring leaf phenology and the diurnal temperature range in a temperate maple forest.

PubMed

Hanes, Jonathan M

2014-03-01

Spring leaf phenology in temperate climates is intricately related to numerous aspects of the lower atmosphere [e.g., surface energy balance, carbon flux, humidity, the diurnal temperature range (DTR)]. To further develop and improve the accuracy of ecosystem and climate models, additional investigations of the specific nature of the relationships between spring leaf phenology and various ecosystem and climate processes are required in different environments. This study used visual observations of maple leaf phenology, below-canopy light intensities, and micrometeorological data collected during the spring seasons of 2008, 2009, and 2010 to examine the potential influence of leaf phenology on a seasonal transition in the trend of the DTR. The timing of a reversal in the DTR trend occurred near the time when the leaves were unfolding and expanding. The results suggest that the spring decline in the DTR can be attributed primarily to the effect of canopy closure on daily maximum temperature. These findings improve our understanding of the relationship between leaf phenology and the diurnal temperature range in temperate maple forests during the spring. They also demonstrate the necessity of incorporating accurate phenological data into ecosystem and climate models and warrant a careful examination of the extent to which canopy phenology is currently incorporated into existing models.

Larger temperature response of autumn leaf senescence than spring leaf-out phenology.

PubMed

Fu, Yongshuo H; Piao, Shilong; Delpierre, Nicolas; Hao, Fanghua; Hänninen, Heikki; Liu, Yongjie; Sun, Wenchao; Janssens, Ivan A; Campioli, Matteo

2018-05-01

Climate warming is substantially shifting the leaf phenological events of plants, and thereby impacting on their individual fitness and also on the structure and functioning of ecosystems. Previous studies have largely focused on the climate impact on spring phenology, and to date the processes underlying leaf senescence and their associated environmental drivers remain poorly understood. In this study, experiments with temperature gradients imposed during the summer and autumn were conducted on saplings of European beech to explore the temperature responses of leaf senescence. An additional warming experiment during winter enabled us to assess the differences in temperature responses of spring leaf-out and autumn leaf senescence. We found that warming significantly delayed the dates of leaf senescence both during summer and autumn warming, with similar temperature sensitivities (6-8 days delay per °C warming), suggesting that, in the absence of water and nutrient limitation, temperature may be a dominant factor controlling the leaf senescence in European beech. Interestingly, we found a significantly larger temperature response of autumn leaf senescence than of spring leaf-out. This suggests a possible larger contribution of delays in autumn senescence, than of the advancement in spring leaf-out, to extending the growing season under future warmer conditions. © 2017 John Wiley & Sons Ltd.

High-resolution prediction of leaf onset date in Japan in the 21st century under the IPCC A1B scenario.

PubMed

Hadano, Mayumi; Nasahara, Kenlo Nishida; Motohka, Takeshi; Noda, Hibiki Muraoka; Murakami, Kazutaka; Hosaka, Masahiro

2013-06-01

Reports indicate that leaf onset (leaf flush) of deciduous trees in cool-temperate ecosystems is occurring earlier in the spring in response to global warming. In this study, we created two types of phenology models, one driven only by warmth (spring warming [SW] model) and another driven by both warmth and winter chilling (parallel chill [PC] model), to predict such phenomena in the Japanese Islands at high spatial resolution (500 m). We calibrated these models using leaf onset dates derived from satellite data (Terra/MODIS) and in situ temperature data derived from a dense network of ground stations Automated Meteorological Data Acquisition System. We ran the model using future climate predictions created by the Japanese Meteorological Agency's MRI-AGCM3.1S model. In comparison to the first decade of the 2000s, our results predict that the date of leaf onset in the 2030s will advance by an average of 12 days under the SW model and 7 days under the PC model throughout the study area. The date of onset in the 2090s will advance by 26 days under the SW model and by 15 days under the PC model. The greatest impact will occur on Hokkaido (the northernmost island) and in the central mountains.

Stabilization of Particle Discrimination Efficiencies for Neutron Spectrum Unfolding With Organic Scintillators

NASA Astrophysics Data System (ADS)

Lawrence, Chris C.; Polack, J. K.; Febbraro, Michael; Kolata, J. J.; Flaska, Marek; Pozzi, S. A.; Becchetti, F. D.

2017-02-01

The literature discussing pulse-shape discrimination (PSD) in organic scintillators dates back several decades. However, little has been written about PSD techniques that are optimized for neutron spectrum unfolding. Variation in n-γ misclassification rates and in γ/n ratio of incident fields can distort the neutron pulse-height response of scintillators and these distortions can in turn cause large errors in unfolded spectra. New applications in arms-control verification call for detection of lower-energy neutrons, for which PSD is particularly problematic. In this article, we propose techniques for removing distortions on pulse-height response that result from the merging of PSD distributions in the low-pulse-height region. These techniques take advantage of the repeatable shapes of PSD distributions that are governed by the counting statistics of scintillation-photon populations. We validate the proposed techniques using accelerator-based time-of-flight measurements and then demonstrate them by unfolding the Watt spectrum from measurement with a 252Cf neutron source.

Insect mimicry of plants dates back to the Permian

PubMed Central

Garrouste, Romain; Hugel, Sylvain; Jacquelin, Lauriane; Rostan, Pierre; Steyer, J.-Sébastien; Desutter-Grandcolas, Laure; Nel, André

2016-01-01

In response to predation pressure, some insects have developed spectacular plant mimicry strategies (homomorphy), involving important changes in their morphology. The fossil record of plant mimicry provides clues to the importance of predation pressure in the deep past. Surprisingly, to date, the oldest confirmed records of insect leaf mimicry are Mesozoic. Here we document a crucial step in the story of adaptive responses to predation by describing a leaf-mimicking katydid from the Middle Permian. Our morphometric analysis demonstrates that leaf-mimicking wings of katydids can be morphologically characterized in a non-arbitrary manner and shows that the new genus and species Permotettigonia gallica developed a mimicking pattern of forewings very similar to those of the modern leaf-like katydids. Our finding suggests that predation pressure was already high enough during the Permian to favour investment in leaf mimicry. PMID:27996977

Insect mimicry of plants dates back to the Permian.

PubMed

Garrouste, Romain; Hugel, Sylvain; Jacquelin, Lauriane; Rostan, Pierre; Steyer, J-Sébastien; Desutter-Grandcolas, Laure; Nel, André

2016-12-20

In response to predation pressure, some insects have developed spectacular plant mimicry strategies (homomorphy), involving important changes in their morphology. The fossil record of plant mimicry provides clues to the importance of predation pressure in the deep past. Surprisingly, to date, the oldest confirmed records of insect leaf mimicry are Mesozoic. Here we document a crucial step in the story of adaptive responses to predation by describing a leaf-mimicking katydid from the Middle Permian. Our morphometric analysis demonstrates that leaf-mimicking wings of katydids can be morphologically characterized in a non-arbitrary manner and shows that the new genus and species Permotettigonia gallica developed a mimicking pattern of forewings very similar to those of the modern leaf-like katydids. Our finding suggests that predation pressure was already high enough during the Permian to favour investment in leaf mimicry.

Insect mimicry of plants dates back to the Permian

NASA Astrophysics Data System (ADS)

Garrouste, Romain; Hugel, Sylvain; Jacquelin, Lauriane; Rostan, Pierre; Steyer, J.-Sébastien; Desutter-Grandcolas, Laure; Nel, André

2016-12-01

In response to predation pressure, some insects have developed spectacular plant mimicry strategies (homomorphy), involving important changes in their morphology. The fossil record of plant mimicry provides clues to the importance of predation pressure in the deep past. Surprisingly, to date, the oldest confirmed records of insect leaf mimicry are Mesozoic. Here we document a crucial step in the story of adaptive responses to predation by describing a leaf-mimicking katydid from the Middle Permian. Our morphometric analysis demonstrates that leaf-mimicking wings of katydids can be morphologically characterized in a non-arbitrary manner and shows that the new genus and species Permotettigonia gallica developed a mimicking pattern of forewings very similar to those of the modern leaf-like katydids. Our finding suggests that predation pressure was already high enough during the Permian to favour investment in leaf mimicry.

High-resolution prediction of leaf onset date in Japan in the 21st century under the IPCC A1B scenario

PubMed Central

Hadano, Mayumi; Nasahara, Kenlo Nishida; Motohka, Takeshi; Noda, Hibiki Muraoka; Murakami, Kazutaka; Hosaka, Masahiro

2013-01-01

Reports indicate that leaf onset (leaf flush) of deciduous trees in cool-temperate ecosystems is occurring earlier in the spring in response to global warming. In this study, we created two types of phenology models, one driven only by warmth (spring warming [SW] model) and another driven by both warmth and winter chilling (parallel chill [PC] model), to predict such phenomena in the Japanese Islands at high spatial resolution (500 m). We calibrated these models using leaf onset dates derived from satellite data (Terra/MODIS) and in situ temperature data derived from a dense network of ground stations Automated Meteorological Data Acquisition System. We ran the model using future climate predictions created by the Japanese Meteorological Agency's MRI-AGCM3.1S model. In comparison to the first decade of the 2000s, our results predict that the date of leaf onset in the 2030s will advance by an average of 12 days under the SW model and 7 days under the PC model throughout the study area. The date of onset in the 2090s will advance by 26 days under the SW model and by 15 days under the PC model. The greatest impact will occur on Hokkaido (the northernmost island) and in the central mountains. PMID:23789086

The leaf phenophase of deciduous species altered by land pavements

NASA Astrophysics Data System (ADS)

Chen, Yuanyuan; Wang, Xiaoke; Jiang, Bo; Li, Li

2018-06-01

It has been widely reported that the urban environment alters leaf and flowering phenophases; however, it remains unclear if land pavement is correlated with these alterations. In this paper, two popular deciduous urban trees in northern China, ash ( Fraxinus chinensis) and maple ( Acer truncatum), were planted in pervious and impervious pavements at three spacings (0.5 m × 0.5 m, 1.0 m × 1.0 m, and 2.0 m × 2.0 m apart). The beginning and end dates of the processes of leaf budburst and senescence were recorded in spring and fall of 2015, respectively. The results show that leaf budburst and senescence were significantly advanced in pavement compared to non-pavement lands. The date of full leaf budburst was earlier by 0.7-9.3 days for ash and by 0.3-2.3 days for maple under pavements than non-pavements, respectively. As tree spacing increases, the advanced days of leaf budburst became longer. Our results clearly indicate that alteration of leaf phenophases is attributed to land pavement, which should be taken into consideration in urban planning and urban plant management.

Effects of forest management practices in temperate beech forests on bacterial and fungal communities involved in leaf litter degradation.

PubMed

Purahong, Witoon; Kapturska, Danuta; Pecyna, Marek J; Jariyavidyanont, Katalee; Kaunzner, Jennifer; Juncheed, Kantida; Uengwetwanit, Tanaporn; Rudloff, Renate; Schulz, Elke; Hofrichter, Martin; Schloter, Michael; Krüger, Dirk; Buscot, François

2015-05-01

Forest management practices (FMPs) significantly influence important ecological processes and services in Central European forests, such as leaf litter decomposition and nutrient cycling. Changes in leaf litter diversity, and thus, its quality as well as microbial community structure and function induced by different FMPs were hypothesized to be the main drivers causing shifts in decomposition rates and nutrient release in managed forests. In a litterbag experiment lasting 473 days, we aimed to investigate the effects of FMPs (even-aged timber management, selective logging and unmanaged) on bacterial and fungal communities involved in leaf litter degradation over time. Our results showed that microbial communities in leaf litter were strongly influenced by both FMPs and sampling date. The results from nonmetric multidimensional scaling (NMDS) ordination revealed distinct patterns of bacterial and fungal successions over time in leaf litter. We demonstrated that FMPs and sampling dates can influence a range of factors, including leaf litter quality, microbial macronutrients, and pH, which significantly correlate with microbial community successions.

The leaf phenophase of deciduous species altered by land pavements

NASA Astrophysics Data System (ADS)

Chen, Yuanyuan; Wang, Xiaoke; Jiang, Bo; Li, Li

2018-02-01

It has been widely reported that the urban environment alters leaf and flowering phenophases; however, it remains unclear if land pavement is correlated with these alterations. In this paper, two popular deciduous urban trees in northern China, ash (Fraxinus chinensis) and maple (Acer truncatum), were planted in pervious and impervious pavements at three spacings (0.5 m × 0.5 m, 1.0 m × 1.0 m, and 2.0 m × 2.0 m apart). The beginning and end dates of the processes of leaf budburst and senescence were recorded in spring and fall of 2015, respectively. The results show that leaf budburst and senescence were significantly advanced in pavement compared to non-pavement lands. The date of full leaf budburst was earlier by 0.7-9.3 days for ash and by 0.3-2.3 days for maple under pavements than non-pavements, respectively. As tree spacing increases, the advanced days of leaf budburst became longer. Our results clearly indicate that alteration of leaf phenophases is attributed to land pavement, which should be taken into consideration in urban planning and urban plant management.

Variation in leaf flushing date influences autumnal senescence and next year’s flushing date in two temperate tree species

PubMed Central

Fu, Yongshuo S. H.; Campioli, Matteo; Vitasse, Yann; De Boeck, Hans J.; Van den Berge, Joke; AbdElgawad, Hamada; Asard, Han; Piao, Shilong; Deckmyn, Gaby; Janssens, Ivan A.

2014-01-01

Recent temperature increases have elicited strong phenological shifts in temperate tree species, with subsequent effects on photosynthesis. Here, we assess the impact of advanced leaf flushing in a winter warming experiment on the current year’s senescence and next year’s leaf flushing dates in two common tree species: Quercus robur L. and Fagus sylvatica L. Results suggest that earlier leaf flushing translated into earlier senescence, thereby partially offsetting the lengthening of the growing season. Moreover, saplings that were warmed in winter–spring 2009–2010 still exhibited earlier leaf flushing in 2011, even though the saplings had been exposed to similar ambient conditions for almost 1 y. Interestingly, for both species similar trends were found in mature trees using a long-term series of phenological records gathered from various locations in Europe. We hypothesize that this long-term legacy effect is related to an advancement of the endormancy phase (chilling phase) in response to the earlier autumnal senescence. Given the importance of phenology in plant and ecosystem functioning, and the prediction of more frequent extremely warm winters, our observations and postulated underlying mechanisms should be tested in other species. PMID:24799708

Variation in leaf flushing date influences autumnal senescence and next year's flushing date in two temperate tree species.

PubMed

Fu, Yongshuo S H; Campioli, Matteo; Vitasse, Yann; De Boeck, Hans J; Van den Berge, Joke; AbdElgawad, Hamada; Asard, Han; Piao, Shilong; Deckmyn, Gaby; Janssens, Ivan A

2014-05-20

Recent temperature increases have elicited strong phenological shifts in temperate tree species, with subsequent effects on photosynthesis. Here, we assess the impact of advanced leaf flushing in a winter warming experiment on the current year's senescence and next year's leaf flushing dates in two common tree species: Quercus robur L. and Fagus sylvatica L. Results suggest that earlier leaf flushing translated into earlier senescence, thereby partially offsetting the lengthening of the growing season. Moreover, saplings that were warmed in winter-spring 2009-2010 still exhibited earlier leaf flushing in 2011, even though the saplings had been exposed to similar ambient conditions for almost 1 y. Interestingly, for both species similar trends were found in mature trees using a long-term series of phenological records gathered from various locations in Europe. We hypothesize that this long-term legacy effect is related to an advancement of the endormancy phase (chilling phase) in response to the earlier autumnal senescence. Given the importance of phenology in plant and ecosystem functioning, and the prediction of more frequent extremely warm winters, our observations and postulated underlying mechanisms should be tested in other species.

Variations in the temperature sensitivity of spring leaf phenology from 1978 to 2014 in Mudanjiang, China

NASA Astrophysics Data System (ADS)

Dai, Junhu; Xu, Yunjia; Wang, Huanjiong; Alatalo, Juha; Tao, Zexing; Ge, Quansheng

2017-12-01

Continuous long-term temperature sensitivity (ST) of leaf unfolding date (LUD) and main impacting factors in spring in the period 1978-2014 for 40 plant species in Mudanjiang, Heilongjiang Province, Northeast China, were analyzed by using observation data from the China Phenological Observation Network (CPON), together with the corresponding meteorological data from the China Meteorological Data Service Center. Temperature sensitivities, slopes of the regression between LUD and mean temperature during the optimum preseason (OP), were analyzed using 15-year moving window to determine their temporal trends. Major factors impacting ST were then chosen and evaluated by applying a random sampling method. The results showed that LUD was sensitive to mean temperature in a defined period before phenophase onset for all plant species analyzed. Over the period 1978-2014, the mean ST of LUD for all plant species was - 3.2 ± 0.49 days °C-1. The moving window analysis revealed that 75% of species displayed increasing ST of LUD, with 55% showing significant increases (P < 0.05). ST for the other 25% exhibited a decreasing trend, with 17% showing significant decreases (P < 0.05). On average, ST increased by 16%, from - 2.8 ± 0.83 days °C-1 during 1980-1994 to - 3.30 ± 0.65 days °C-1 during 2000-2014. For species with later LUD and longer OP, ST tended to increase more, while species with earlier LUD and shorter OP tended to display a decreasing ST. The standard deviation of preseason temperature impacted the temporal variation in ST. Chilling conditions influenced ST for some species, but photoperiod limitation did not have significant or coherent effects on changes in ST.

76 FR 76297 - Drawbridge Operation Regulation; Willamette River, Portland, OR

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-07

.... This deviation allows single leaf operations during rehabilitation and requires a two hour advance notification for a single leaf opening. DATES: This deviation is effective from 6 a.m. on December 1, 2011...: Multnomah County has requested to operate the Morrison Bascule Bridge with single leaf openings with two...

[An early warning method of cucumber downy mildew in solar greenhouse based on canopy temperature and humidity modeling].

PubMed

Wang, Hui; Li, Mei-lan; Xu, Jian-ping; Chen, Mei-xiang; Li, Wen-yong; Li, Ming

2015-10-01

The greenhouse environmental parameters can be used to establish greenhouse nirco-climate model, which can combine with disease model for early warning, with aim of ecological controlling diseases to reduce pesticide usage, and protecting greenhouse ecological environment to ensure the agricultural product quality safety. Greenhouse canopy leaf temperature and air relative humidity, models were established using energy balance and moisture balance principle inside the greenhouse. The leaf temperature model considered radiation heat transfer between the greenhouse crops, wall, soil and cover, plus the heat exchange caused by indoor net radiation and crop transpiration. Furthermore, the water dynamic balance in the greenhouse including leaf transpiration, soil evaporation, cover and leaf water vapor condensation, was considered to develop a relative humidity model. The primary infection and latent period warning models for cucumber downy mildew (Pseudoperonospora cubensis) were validated using the results of the leaf temperature and relative humidity model, and then the estimated disease occurrence date of cucumber downy mildew was compared with actual disease occurrence date of field observation. Finally, the results were verified by the measured temperature and humidity data of September and October, 2014. The results showed that the root mean square deviations (RMSDs) of the measured and estimated leaf temperature were 0.016 and 0.024 °C, and the RMSDs of the measured and estimated air relative humidity were 0.15% and 0.13%, respectively. Combining the result of estimated temperature and humidity models, a cucumber disease early warning system was established to forecast the date of disease occurrence, which met with the real date. Thus, this work could provide the micro-environment data for the early warning system of cucumber diseases in solar greenhouses.

Field evidence for earlier leaf-out dates in alpine grassland on the eastern Tibetan Plateau from 1990 to 2006.

PubMed

Zhou, H K; Yao, B Q; Xu, W X; Ye, X; Fu, J J; Jin, Y X; Zhao, X Q

2014-08-01

Worldwide, many plant species are experiencing an earlier onset of spring phenophases due to climate warming. Rapid recent temperature increases on the Tibetan Plateau (TP) have triggered changes in the spring phenology of the local vegetation. However, remote sensing studies of the land surface phenology have reached conflicting interpretations about green-up patterns observed on the TP since the mid-1990s. We investigated this issue using field phenological observations from 1990 to 2006, for 11 dominant plants on the TP at the levels of species, families (Gramineae-grasses and Cyperaceae-sedges) and vegetation communities (alpine meadow and alpine steppe). We found a significant trend of earlier leaf-out dates for one species (Koeleria cristata). The leaf-out dates of both Gramineae and Cyperaceae had advanced (the latter significantly, starting an average of 9 days later per year than the former), but the correlation between them was significant. The leaf-out dates of both vegetation communities also advanced, but the pattern was only significant in the alpine meadow. This study provides the first field evidence of advancement in spring leaf phenology on the TP and suggests that the phenology of the alpine steppe can differ from that of the alpine meadow. These findings will be useful for understanding ecosystem responses to climate change and for grassland management on the TP. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Evaluating Gridded Spring Indices Using the USA National Phenology Network's Observational Phenology Data

NASA Astrophysics Data System (ADS)

Crimmins, T. M.; Gerst, K.

2017-12-01

The USA National Phenology Network (USA-NPN; www.usanpn.org) produces and freely delivers daily and short-term forecast maps of spring onset dates at fine spatial scale for the conterminous United States and Alaska using the Spring Indices. These models, which represent the start of biological activity in the spring season, were developed using a long-term observational record of four species of lilacs and honeysuckles contributed by volunteer observers. Three of the four species continue to be tracked through the USA-NPN's phenology observation program, Nature's Notebook. The gridded Spring Index maps have utility for a wide range of natural resource planning and management applications, including scheduling invasive species and pest detection and control activities, anticipating allergy outbreaks and planning agricultural harvest dates. However, to date, there has not been a comprehensive assessment of how well the gridded Spring Index maps accurately reflect phenological activity in lilacs and honeysuckles or other species of plants. In this study, we used observational plant phenology data maintained by the USA-NPN to evaluate how well the gridded Spring Index maps match leaf and flowering onset dates in a) the lilac and honeysuckle species used to construct the models and b) in several species of deciduous trees. The Spring Index performed strongly at predicting the timing of leaf-out and flowering in lilacs and honeysuckles. The average error between predicted and observed date of onset ranged from 5.9 to 11.4 days. Flowering models performed slightly better than leaf-out models. The degree to which the Spring Indices predicted native deciduous tree leaf and flower phenology varied by year, species, and region. Generally, the models were better predictors of leaf and flowering onset dates in the Northeastern and Midwestern US. These results reveal when and where the Spring Indices are a meaningful proxy of phenological activity across the United States.

Leaf sample detail, Feb2016-May2016, PA-SLZ, PA-PNM, PA-BCI: Panama

DOE Data Explorer

Ely, Kim [Brookhaven National Lab; Rogers, Alistair [Brookhaven National Lab; Serbin, Shawn [Brookhaven National Lab; Wu, Jin [BNL; Wolfe, Brett [Smithsonian; Dickman, Turin [Los Alamos National Lab; Collins, Adam [Los Alamos National Lab; Detto, Matteo [Princeton; Grossiord, Charlotte [Los Alamos National Lab; McDowell, Nate [Los Alamos National Lab; Michaletz, Sean

2017-01-01

Date, location, species and photographs of leaf samples collected on a monthly basis from Feb to May 2016 at SLZ and PNM. Data from BCI only available for March. This data was collected as part of the 2016 ENSO campaign. Data to be used as a reference to linking related datasets (existing and future) including leaf water potential, leaf spectra, LMA, gas exchange and leaf chemistry (CHN, NSC, P). Most leaves were sampled from sunlit canopy trees.

Model estimates of leaf area and reference canopy stomatal conductance suggest correlation between phenology and physiology in both trembling aspen and red pine

NASA Astrophysics Data System (ADS)

Mackay, D. S.; Ewers, B. E.; Kruger, E. L.

2006-12-01

Phenological variations impact water and carbon fluxes, as evidenced by the large interannual variability of net ecosystem exchange of carbon dioxide and evapotranspiration (ET). In northern Wisconsin we observed daily variations of canopy transpiration from hardwoods from 1.0 to 1.7 mm/day during the leaf unfolding period and 1.7 to 2.6 mm/day with leaves fully out. Correlations between such flux rates and phenology have not been extensively tested and mechanistic connections are in their infancy. Some data suggest that stomatal conductance and photosynthesis increases up to full expansion. Moreover, in conifers, the interaction of phenology and physiology is more complicated than in deciduous trees because needles are retained for several years. Using inverse modeling with a coupled photosynthesis-transpiration model we estimated reference canopy stomatal conductance, Gsref, for red pine (Pinus resinosa), and Gsref and leaf area index, L, for trembling aspen (Populus tremuloides), using 30-min continuous sap flux data spanning a period from just prior to the start of leaf expansion to just after leaf senescence. The red pine showed Gsref ramp up from 105 to 179 mmol m-2 leaf s-1, which represented a 37 to 50 percent increase in Gsref after accounting for maximum possible changes in L. After full leaf out, the trembling aspen were almost immediately defoliated, and then reflushed after three weeks. Model estimates of L reflected this pattern and were consistent with measurements. However, Gsref never exceeded 45 mmol m-2 s-1 prior to defoliation, but peaked at 112 mmol m-2 s-1 after reflushing. These results support the need for further work that aims to separate phenology and physiology.

78 FR 5174 - Combined Notice of Filings

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-24

... Existing Proceedings Docket Numbers: RP12-1067-002. Applicants: Leaf River Energy Center LLC. Description: Leaf River Energy Center LLC--Revised Compliance Filing to be effective 12/1/2012. Filed Date: 1/11/13...

77 FR 40509 - Drawbridge Operation Regulation; Oakland Inner Harbor Tidal Canal, Alameda, CA

Federal Register 2010, 2011, 2012, 2013, 2014

2012-07-10

... Agency to perform necessary repairs on the drawbridge. This deviation allows single leaf operation of the double leaf bascule style drawbridge during the project. DATES: This deviation is effective from 7 a.m... leaf of the double bascule drawspan may be secured in the closed-to-navigation position from 7 a.m...

75 FR 1705 - Drawbridge Operation Regulation; Intracoastal Waterway (ICW), Inside Thorofare, Ventnor City, NJ

Federal Register 2010, 2011, 2012, 2013, 2014

2010-01-13

... Avenue Bridge, at ICW mile 71.2, across Inside Thorofare, at Ventnor City. This bridge is a double-leaf... rehabilitation to one of the bascule leafs. DATES: This deviation is effective from 7 a.m. on January 20, 2010... Atlantic County who owns and operates this double-leaf bascule drawbridge, has requested a temporary...

Forecasting plant phenology: evaluating the phenological models for Betula pendula and Padus racemosa spring phases, Latvia.

PubMed

Kalvāns, Andis; Bitāne, Māra; Kalvāne, Gunta

2015-02-01

A historical phenological record and meteorological data of the period 1960-2009 are used to analyse the ability of seven phenological models to predict leaf unfolding and beginning of flowering for two tree species-silver birch Betula pendula and bird cherry Padus racemosa-in Latvia. Model stability is estimated performing multiple model fitting runs using half of the data for model training and the other half for evaluation. Correlation coefficient, mean absolute error and mean squared error are used to evaluate model performance. UniChill (a model using sigmoidal development rate and temperature relationship and taking into account the necessity for dormancy release) and DDcos (a simple degree-day model considering the diurnal temperature fluctuations) are found to be the best models for describing the considered spring phases. A strong collinearity between base temperature and required heat sum is found for several model fitting runs of the simple degree-day based models. Large variation of the model parameters between different model fitting runs in case of more complex models indicates similar collinearity and over-parameterization of these models. It is suggested that model performance can be improved by incorporating the resolved daily temperature fluctuations of the DDcos model into the framework of the more complex models (e.g. UniChill). The average base temperature, as found by DDcos model, for B. pendula leaf unfolding is 5.6 °C and for the start of the flowering 6.7 °C; for P. racemosa, the respective base temperatures are 3.2 °C and 3.4 °C.

Chronological Sequence of Leaf Phenology, Xylem and Phloem Formation and Sap Flow of Quercus pubescens from Abandoned Karst Grasslands

PubMed Central

Lavrič, Martina; Eler, Klemen; Ferlan, Mitja; Vodnik, Dominik; Gričar, Jožica

2017-01-01

Intra-annual variations in leaf development, radial growth, including the phloem part, and sap flow have rarely been studied in deciduous trees from drought-prone environments. In order to understand better the chronological order and temporal course of these processes, we monitored leaf phenology, xylem and phloem formation and sap flow in Quercus pubescens from abandoned karst grasslands in Slovenia during the growing season of 2014. We found that the initial earlywood vessel formation started before bud opening at the beginning of April. Buds started to open in the second half of April and full leaf unfolding occurred by the end of May. LAI values increased correspondingly with leaf development. About 28% of xylem and 22% of phloem annual increment were formed by the time of bud break. Initial earlywood vessels were fully lignified and ready for water transport, indicating that they are essential to provide hydraulic conductivity for axial water flow during leaf development. Sap flow became active and increasing contemporarily with leaf development and LAI values. Similar early spring patterns of xylem sap flow and LAI denoted that water transport in oaks broadly followed canopy leaf area development. In the initial 3 weeks of radial growth, phloem growth preceded that of xylem, indicating its priority over xylem at the beginning of the growing season. This may be related to the fact that after bud break, the developing foliage is a very large sink for carbohydrates but, at the same time, represents a small transpirational area. Whether the interdependence of the chronological sequence of the studied processes is fixed in Q. pubescens needs to be confirmed with more data and several years of analyses, although the ‘correct sequence’ of processes is essential for synchronized plant performance and response to environmental stress. PMID:28321232

Highly Decorated Lignins in Leaf Tissues of the Canary Island Date Palm Phoenix canariensis1[OPEN

PubMed Central

Bartuce, Allison; Free, Heather C.A.; Smith, Bronwen G.

2017-01-01

The cell walls of leaf base tissues of the Canary Island date palm (Phoenix canariensis) contain lignins with the most complex compositions described to date. The lignin composition varies by tissue region and is derived from traditional monolignols (ML) along with an unprecedented range of ML conjugates: ML-acetate, ML-benzoate, ML-p-hydroxybenzoate, ML-vanillate, ML-p-coumarate, and ML-ferulate. The specific functions of such complex lignin compositions are unknown. However, the distribution of the ML conjugates varies depending on the tissue region, indicating that they may play specific roles in the cell walls of these tissues and/or in the plant’s defense system. PMID:28894022

Phenological responses of plants in Europe: Differences in timing due to the type of event, the location and human density

NASA Astrophysics Data System (ADS)

Estrella, N.; Menzel, A.

2009-04-01

The 4th assessment report of the IPCC confirms that there is already a change in biological and physical systems in the expected direction due to climate change. Even the human influence could be verified in these changes. Plant phenological observations provide a good tool to track the quality and quantity of plant reactions to temperature. In Europe, in particular, there has been a long tradition of collecting onset dates for various plant phenological phases. We analysed a huge phenological data set collected during the COST 725 Action ‘Establishing a European Phenological Data Platform for Climatological Applications' containing more than 36,000 phenological time series (between 1971-2000) for Europe. This dataset was analysed by Menzel et al. (2006) on a national basis and they discovered that there was a extensive and apparent change in phenology over the three decades towards earlier start for spring phases and a slightly delayed beginning of leaf colouring in autumn. In contrast to the Menzel et al (2006) study we examined pan-European differences in the behaviour of phenological phases on the basis of station time series. We focussed on the temperature response of phenological phases and their regional patterns and the relationship between local temperature and phenological trends as well as the influence of human population density. The temperature information was derived from a high-resolution climate grid of Europe (0.5° x 0.5° grid CRU TS 2.1 (Mitchell & Jones 2005)). The data on population density were downloaded from Euro Stat. The results of our analyses confirm differences in behaviour between annual and perennial plants in Europe; the temperature response of perennial plants was greater (-4.2 days/°C) than that of annual agricultural crops (-3.0 days/°C). The correlation between the temperature trend and the phenology trend was strongest for leaf unfolding of fruit and deciduous trees (r=-0.63 and r=-0.46 respectively). Geographical coordinates (latitude and longitude) alone only had little impact on the mean onset of the group of phases, including altitude there was an influence for some groups. We could show that human population density influenced the mean onset date of the group of phases.

Monitoring phenology of photosynthesis in temperate evergreen and mixed deciduous forests using the normalized difference vegetation index (NDVI) and the photochemical reflectance index (PRI) at leaf and canopy scales

NASA Astrophysics Data System (ADS)

Wong, C. Y.; Arain, M. A.; Ensminger, I.

2016-12-01

Evergreen conifers in boreal and temperate regions undergo strong seasonal changes in photoperiod and temperatures, which determines their phenology of high photosynthetic activity in the growing season and downregulation during the winter. Monitoring the timing of the transition between summer activity and winter downregulation in evergreens is difficult since this is a largely invisible process, unlike in deciduous trees that have a visible budding and a sequence of leaf unfolding in the spring and leaf abscission in the fall. The light-use efficiency (LUE) model estimates gross primary productivity (GPP) and may be parameterized using remotely sensed vegetation indices. Using spectral reflectance data, we derived the normalized difference vegetation index (NDVI), a measure of leaf "greenness", and the photochemical reflectance index (PRI), a proxy for chlorophyll:carotenoid ratios which is related to photosynthetic activity. To better understand the relationship between these vegetation indices and photosynthetic activity and to contrast this relationship between plant functional types, the phenology of NDVI, PRI and photosynthesis was monitored in an evergreen forest and a mixed deciduous forest at the leaf and canopy scale. Our data indicates that the LUE model can be parameterized by NDVI and PRI to track forest phenology. Differences in the sensitivity of PRI and NDVI will be discussed. These findings have implications to address the phenology of evergreen conifers by using PRI to complement NDVI in the LUE model, potentially improving model productivity estimates in northern hemisphere forests, that are dominated by conifers.

Late Quaternary climate and environmental reconstruction based on leaf wax analyses in the loess sequence of Möhlin, Switzerland

NASA Astrophysics Data System (ADS)

Wüthrich, Lorenz; Bliedtner, Marcel; Kathrin Schäfer, Imke; Zech, Jana; Shajari, Fatemeh; Gaar, Dorian; Preusser, Frank; Salazar, Gary; Szidat, Sönke; Zech, Roland

2017-12-01

We present the results of leaf wax analyses (long-chain n-alkanes) from the 6.8 m deep loess sequence of Möhlin, Switzerland, spanning the last ˜ 70 kyr. Leaf waxes are well preserved and occur in sufficient amounts only down to 0.4 m and below 1.8 m depth, so no paleoenvironmental reconstructions can be done for marine isotope stage (MIS) 2. Compound-specific δ2Hwax analyses yielded similar values for late MIS 3 compared to the uppermost samples, indicating that various effects (e.g., more negative values due to lower temperatures, more positive values due to an enriched moisture source) cancel each other out. A pronounced ˜ 30 ‰ shift towards more negative values probably reflects more humid conditions before ˜ 32 ka. Radiocarbon dating of the n-alkanes corroborates the stratigraphic integrity of leaf waxes and their potential for dating loess-paleosol sequences (LPS) back to ˜ 30 ka.

Responses of canopy duration to temperature changes in four temperate tree species: relative contributions of spring and autumn leaf phenology.

PubMed

Vitasse, Yann; Porté, Annabel Josée; Kremer, Antoine; Michalet, Richard; Delzon, Sylvain

2009-08-01

While changes in spring phenological events due to global warming have been widely documented, changes in autumn phenology, and therefore in growing season length, are less studied and poorly understood. However, it may be helpful to assess the potential lengthening of the growing season under climate warming in order to determine its further impact on forest productivity and C balance. The present study aimed to: (1) characterise the sensitivity of leaf phenological events to temperature, and (2) quantify the relative contributions of leaf unfolding and senescence to the extension of canopy duration with increasing temperature, in four deciduous tree species (Acer pseudoplatanus, Fagus sylvatica, Fraxinus excelsior and Quercus petraea). For 3 consecutive years, we monitored the spring and autumn phenology of 41 populations at elevations ranging from 100 to 1,600 m. Overall, we found significant altitudinal trends in leaf phenology and species-specific differences in temperature sensitivity. With increasing temperature, we recorded an advance in flushing from 1.9 +/- 0.3 to 6.6 +/- 0.4 days degrees C(-1) (mean +/- SD) and a 0 to 5.6 +/- 0.6 days degrees C(-1) delay in leaf senescence. Together both changes resulted in a 6.9 +/- 1.0 to 13.0 +/- 0.7 days degrees C(-1) lengthening of canopy duration depending on species. For three of the four studied species, advances in flushing were the main factor responsible for lengthening canopy duration with increasing temperature, leading to a potentially larger gain in solar radiation than delays in leaf senescence. In contrast, for beech, we found a higher sensitivity to temperature in leaf senescence than in flushing, resulting in an equivalent contribution in solar radiation gain. These results suggest that climate warming will alter the C uptake period and forest productivity by lengthening canopy duration. Moreover, the between-species differences in phenological responses to temperature evidenced here could affect biotic interactions under climate warming.

Enzymatic activities in different strains isolated from healthy and brittle leaf disease affected date palm leaves: study of amylase production conditions.

PubMed

Mouna, Jrad; Imen, Fendri; Choba Ines, Ben; Nourredine, Drira; Adel, Kadri; Néji, Gharsallah

2015-02-01

The present study aimed to investigate and compare the enzymatic production of endophytic bacteria isolated from healthy and brittle leaf disease affected date palm leaves (pectinase, cellulase, lipase, and amylase). The findings revealed that the enzymatic products from the bacterial isolates of healthy date palm leaves were primarily 33% amylolytic enzyme, 33 % cellulase, 25 % pectinase, and 25 % lipase. The isolates from brittle leaf disease date palm leaves, on the other hand, were noted to produce 16 % amylolytic enzyme, 20 % cellulose, 50 % pectinase, and 50 % lipase. The effects of temperature and pH on amylase, pectinase, and cellulose activities were investigated. The Bacillus subtilis JN934392 strain isolated from healthy date palm leaves produced higher levels of amylase activity at pH 7. A Box Behnken Design (BBD) was employed to optimize amylase extraction. Maximal activity was observed at pH and temperature ranges of pH 6-6.5 and 37-39 °C, respectively. Under those conditions, amylase activity was noted to be attained 9.37 U/ml. The results showed that the enzyme was able to maintain more than 50 % of its activity over a temperature range of 50-80 °C, with an optimum at 70 °C. This bacterial amylase showed high activity compared to other bacteria, which provides support for its promising candidacy for future industrial application.

Synthesizing plant phenological indicators from multispecies datasets

NASA Astrophysics Data System (ADS)

Rutishauser, This; Peñuelas, Josep; Filella, Iolanda; Gehrig, Regula; Scherrer, Simon C.; Röthlisberger, Christian

2014-05-01

Changes in the seasonality of life cycles of plants from phenological observations are traditionally analysed at the species level. Trends and correlations with main environmental driving variables show a coherent picture across the globe. The question arises whether there is an integrated phenological signal across species that describes common interannual variability. Is there a way to express synthetic phenological indicators from multispecies datasets that serve decision makers as usefull tools? Can these indicators be derived in such a robust way that systematic updates yield necessary information for adaptation measures? We address these questions by analysing multi-species phenological data sets with leaf-unfolding and flowering observations from 30 sites across Europe between 40° and 63°N including data from PEP725, the Swiss Plant Phenological Observation Network and one legacy data set. Starting in 1951 the data sets were synthesized by multivariate analysis (Principal Component Analysis). The representativeness of the site specific indicator was tested against subsets including only leaf-unfolding or flowering phases, and by a comparison with a 50% random sample of the available phenophases for 500 time steps. Results show that a synthetic indicators explains up to 79% of the variance at each site - usually 40-50% or more. Robust linear trends over the common period 1971-2000 indicate an overall change of the indicator of -0.32 days/year with lower uncertainty than previous studies. Advances were more pronounced in southern and northern Europe. The indicator-based analysis provides a promising tool for synthesizing site-based plant phenological records and is a companion to, and validating data for, an increasing number of phenological measurements derived from phenological models and satellite sensors.

Endophytic colonization of date palm (Phoenix dactylifera L.) leaves by entomopathogenic fungi.

PubMed

Gómez-Vidal, S; Lopez-Llorca, L V; Jansson, H -B; Salinas, J

2006-01-01

Light and scanning electron microscopy together with fungal isolation techniques were used to detect entomopathogenic fungi within young and adult date palm (Phoenix dactylifera) petioles and to assess fungal survival in leaf tissues. The entomopathogenic fungi Beauveria bassiana, Lecanicillium dimorphum and Lecanicillium c.f. psalliotae survived inside leaf tissues at least 30 days after inoculation. Entomopathogenic fungi colonized inoculated petioles endophytically and were recovered up to 3cm from the inoculation site. Fungi were detected inside the parenchyma and sparsely within vascular tissue using microscopy techniques. Our results show that the entomopathogenic fungi used in this study survived and colonized date palm tissues in bioassays both under laboratory and field experimental conditions with no evidence of significant damage.

Arabidopsis thaliana alpha1,2-glucosyltransferase (ALG10) is required for efficient N-glycosylation and leaf growth

PubMed Central

Farid, Akhlaq; Pabst, Martin; Schoberer, Jennifer; Altmann, Friedrich; Glössl, Josef; Strasser, Richard

2011-01-01

Assembly of the dolichol-linked oligosaccharide precursor (Glc3Man9GlcNAc2) is highly conserved among eukaryotes. In contrast to yeast and mammals, little is known about the biosynthesis of dolichol-linked oligosaccharides and the transfer to asparagine residues of nascent polypeptides in plants. To understand the biological function of these processes in plants we characterized the Arabidopsis thaliana homolog of yeast ALG10, the α1,2-glucosyltransferase that transfers the terminal glucose residue to the lipid-linked precursor. Expression of an Arabidopsis ALG10–GFP fusion protein in Nicotiana benthamiana leaf epidermal cells revealed a reticular distribution pattern resembling endoplasmic reticulum (ER) localization. Analysis of lipid-linked oligosaccharides showed that Arabidopsis ALG10 can complement the yeast Δalg10 mutant strain. A homozygous Arabidopsis T-DNA insertion mutant (alg10-1) accumulated mainly lipid-linked Glc2Man9GlcNAc2 and displayed a severe protein underglycosylation defect. Phenotypic analysis of alg10-1 showed that mutant plants have altered leaf size when grown in soil. Moreover, the inactivation of ALG10 in Arabidopsis resulted in the activation of the unfolded protein response, increased salt sensitivity and suppression of the phenotype of α-glucosidase I-deficient plants. In summary, these data show that Arabidopsis ALG10 is an ER-resident α1,2-glucosyltransferase that is required for lipid-linked oligosaccharide biosynthesis and subsequently for normal leaf development and abiotic stress response. PMID:21707802

Human Rights: Unfolding of the American Tradition. Report No. 8403.

ERIC Educational Resources Information Center

Bureau of Public Affairs (Dept. of State), Washington, DC.

Excerpts from 100 speeches, essays, and legal documents dating from classical times to the present illustrate the record of human rights discussion over the centuries. The compilation was made in 1968 to mark the 20th anniversary of the Universal Declaration of Human Rights. The readings indicate that human rights initially meant freedom from a…

Evidence for the principle of minimal frustration in the evolution of protein folding landscapes.

PubMed

Tzul, Franco O; Vasilchuk, Daniel; Makhatadze, George I

2017-02-28

Theoretical and experimental studies have firmly established that protein folding can be described by a funneled energy landscape. This funneled energy landscape is the result of foldable protein sequences evolving following the principle of minimal frustration, which allows proteins to rapidly fold to their native biologically functional conformations. For a protein family with a given functional fold, the principle of minimal frustration suggests that, independent of sequence, all proteins within this family should fold with similar rates. However, depending on the optimal living temperature of the organism, proteins also need to modulate their thermodynamic stability. Consequently, the difference in thermodynamic stability should be primarily caused by differences in the unfolding rates. To test this hypothesis experimentally, we performed comprehensive thermodynamic and kinetic analyses of 15 different proteins from the thioredoxin family. Eight of these thioredoxins were extant proteins from psychrophilic, mesophilic, or thermophilic organisms. The other seven protein sequences were obtained using ancestral sequence reconstruction and can be dated back over 4 billion years. We found that all studied proteins fold with very similar rates but unfold with rates that differ up to three orders of magnitude. The unfolding rates correlate well with the thermodynamic stability of the proteins. Moreover, proteins that unfold slower are more resistant to proteolysis. These results provide direct experimental support to the principle of minimal frustration hypothesis.

Genetic variation in transpiration efficiency and relationships between whole plant and leaf gas exchange measurements in Saccharum spp. and related germplasm.

PubMed

Jackson, Phillip; Basnayake, Jaya; Inman-Bamber, Geoff; Lakshmanan, Prakash; Natarajan, Sijesh; Stokes, Chris

2016-02-01

Fifty-one genotypes of sugarcane (Saccharum spp.) or closely related germplasm were evaluated in a pot experiment to examine genetic variation in transpiration efficiency. Significant variation in whole plant transpiration efficiency was observed, with the difference between lowest and highest genotypes being about 40% of the mean. Leaf gas exchange measurements were made across a wide range of conditions. There was significant genetic variation in intrinsic transpiration efficiency at a leaf level as measured by leaf internal CO2 (Ci) levels. Significant genetic variation in Ci was also observed within subsets of data representing narrow ranges of stomatal conductance. Ci had a low broad sense heritability (Hb = 0.11) on the basis of single measurements made at particular dates, because of high error variation and genotype × date interaction, but broad sense heritability for mean Ci across all dates was high (Hb = 0.81) because of the large number of measurements taken at different dates. Ci levels among genotypes at mid-range levels of conductance had a strong genetic correlation (-0.92 ± 0.30) with whole plant transpiration efficiency but genetic correlations between Ci and whole plant transpiration efficiency were weaker or not significant at higher and lower levels of conductance. Reduced Ci levels at any given level of conductance may result in improved yields in water-limited environments without trade-offs in rates of water use and growth. Targeted selection and improvement of lowered Ci per unit conductance via breeding may provide longer-term benefits for water-limited environments but the challenge will be to identify a low-cost screening methodology. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Determining density of maize canopy. 3: Temporal considerations

NASA Technical Reports Server (NTRS)

Stoner, E. R.; Baumgardner, M. F.; Anuta, P. E.; Cipra, J. E.

1972-01-01

Multispectral scanner data were collected in two flights over ground cover plots at an altitude of 305 m. Eight ground reflectance panels in close proximity to the ground cover plots were used to normalize the scanner data obtained on different dates. Separate prediction equations were obtained for both flight dates for all eleven reflective wavelength bands of the multispectral scanner. Ratios of normalized scanner data were related to leaf area index over time. Normalized scanner data were used to plot relative reflectance versus wavelength for the ground cover plots. Spectral response curves were similar to those for bare soil and green vegetation as determined by laboratory measurements. The spectral response curves from the normalized scanner data indicated that reflectance in the 0.72 to 1.3 micron wavelength range increased as leaf area index increased. A decrease in reflectance was observed in the 0.65 micron chlorophyll absorption band as leaf area index increased.

Determining density of maize canopy. 2: Airborne multispectral scanner data

NASA Technical Reports Server (NTRS)

Stoner, E. R.; Baumgardner, M. F.; Cipra, J. E.

1971-01-01

Multispectral scanner data were collected in two flights over a light colored soil background cover plot at an altitude of 305 m. Energy in eleven reflective wavelength band from 0.45 to 2.6 microns was recorded. Four growth stages of maize (Zea mays L.) gave a wide range of canopy densities for each flight date. Leaf area index measurements were taken from the twelve subplots and were used as a measure of canopy density. Ratio techniques were used to relate uncalibrated scanner response to leaf area index. The ratios of scanner data values for the 0.72 to 0.92 micron wavelength band over the 0.61 to 0.70 micron wavelength band were calculated for each plot. The ratios related very well to leaf area index for a given flight date. The results indicated that spectral data from maize canopies could be of value in determining canopy density.

Antiquity of coca-leaf chewing in the south central Andes: a 3,000 year archaeological record of coca-leaf chewing from northern Chile.

PubMed

Rivera, Mario A; Aufderheide, Arthur C; Cartmell, Larry W; Torres, Constantino M; Langsjoen, Odin

2005-12-01

Carbon-14 (14C) dating from mummies of the Alto Ramirez culture confirms that coca leaf chewing was an incipient practice among members of a population that peopled the valleys and coastal areas of Northern Chile by 3,000 years before the present (yr.B.P.). Out of eleven bodies from the burial site of Pisagua-7 (PSG-7, S 19 degrees 35', W 70 degrees 13') that were analyzed, two samples tested positive. Mummy 725-A C2 (dated 3,090 to 2,850 two sigma calibrated 14C years before the present) was shown to have a cocaine value of 13.3 nanograms/10 milligrams of sample (ng/10mg), and mummy 741 (2,890 to 2,760 two sigma cal yr B.P.), a 5.6 ng/10mg value.

Bud Composition, Branching Patterns and Leaf Phenology in Cerrado Woody Species

PubMed Central

DAMASCOS, M. A.; PRADO, C. H. B. A.; RONQUIM, C. C.

2005-01-01

• Background and Aims Plants have complex mechanisms of aerial biomass exposition, which depend on bud composition, the period of the year in which shoot extension occurs, branching pattern, foliage persistence, herbivory and environmental conditions. • Methods The influence of water availability and temperature on shoot growth, the bud composition, the leaf phenology, and the relationship between partial leaf fall and branching were evaluated over 3 years in Cerrado woody species Bauhinia rufa (BR), Leandra lacunosa (LL) and Miconia albicans (MA). • Key Results Deciduous BR preformed organs in buds and leaves flush synchronously at the transition from the dry to the wet season. The expansion time of leaves is <1 month. Main shoots (first-order axis, A1 shoots) extended over 30 d and they did not branch. BR budding and foliage unfolds were brought about independently of inter-annual rainfall variations. By contrast, in LL and MA evergreen species, the shoot extension rate and the neoformation of aerial organs depended on rainfall. Leaf emergence was continuous for 2–6 months and lamina expansion took place over 1–4 months. The leaf life span was 5–20 months and the main A1 shoot extension happened over 122–177 d. Both evergreen species allocated biomass to shoots, leaves or flowers continuously during the year, branching in the middle of the wet season to form second-order (A2 shoots) and third-order (A3 shoots) axis in LL and A2 shoots in MA. Partial shed of A1 shoot leaves would facilitate a higher branching intensity A2 shoot production in LL than in MA. MA presented a longer leaf life span, produced a lower percentage of A2 shoots but had a higher meristem persistence on A1 and A2 shoots than LL. • Conclusions It was possible to identify different patterns of aerial growth in Cerrado woody species defined by shoot-linked traits such as branching pattern, bud composition, meristem persistence and leaf phenology. These related traits must be considered over and above leaf deciduousness for searching functional guilds in a Cerrado woody community. For the first time a relationship between bud composition, shoot growth and leaf production pattern is found in savanna woody plants. PMID:16157631

Survival and development of Heliothis virescens (Lepidoptera: Noctuidae) larvae on isogenic tobacco lines with different levels of alkaloids.

PubMed

Jackson, D Michael; Johnson, A W; Stephenson, M G

2002-12-01

Levels of pyridine alkaloids were measured in 18 tobacco, Nicotiana tabacum L., entries from three parental isolines ('NC 95', 'SC 58', and 'Coker 139'), grown at Tifton, GA, Florence, SC, and Oxford, NC, in 1991. Levels of alkaloids in bud leaves (first fully unfolded leaf below the apical leaf bud) were negatively correlated to natural infestation ratings of tobacco budworm larvae, Heliothis virescens (F.), 7 wk after transplanting. For artificially infested bud leaves at Oxford, there was a significant negative correlation between levels of total alkaloids and larval weights after 1 wk of feeding. In 1992, four entries from the 'NC 95' isoline were grown at Oxford, and samples for alkaloid analyses were taken every 2 wk at several leaf positions on each plant. During weeks 4, 8, 12, and 16, second instar tobacco budworms were caged on individual, intact leaves inside perforated plastic bags in the field. The survival and development of tobacco budworm larvae after 1 wk were negatively correlated with levels of alkaloids at the various leaf positions. Larvae survived better and grew faster on the bud leaves of each entry where alkaloid levels were lower than they did on leaves further down the stalk where alkaloid levels were higher. More larvae survived on the lower leaves of the low alkaloid lines than on the lower leaves of the high alkaloid lines. Even moderate increases in pyridine alkaloids had negative effects on tobacco budworm survival and development. Nicotine constituted >97% of the pyridine alkaloids in the 'NC95' isoline each year.

Mechanical Properties of β-Catenin Revealed by Single-Molecule Experiments

PubMed Central

Valbuena, Alejandro; Vera, Andrés Manuel; Oroz, Javier; Menéndez, Margarita; Carrión-Vázquez, Mariano

2012-01-01

β-catenin is a central component of the adaptor complex that links cadherins to the actin cytoskeleton in adherens junctions and thus, it is a good candidate to sense and transmit mechanical forces to trigger specific changes inside the cell. To fully understand its molecular physiology, we must first investigate its mechanical role in mechanotransduction within the cadherin system. We have studied the mechanical response of β-catenin to stretching using single-molecule force spectroscopy and molecular dynamics. Unlike most proteins analyzed to date, which have a fixed mechanical unfolding pathway, the β-catenin armadillo repeat region (ARM) displays low mechanostability and multiple alternative unfolding pathways that seem to be modulated by its unstructured termini. These results are supported by steered molecular dynamics simulations, which also predict its mechanical stabilization and unfolding pathway restrictions when the contiguous α-helix of the C-terminal unstructured region is included. Furthermore, simulations of the ARM/E-cadherin cytosolic tail complex emulating the most probable stress geometry occurring in vivo show a mechanical stabilization of the interaction whose magnitude correlates with the length of the stretch of the cadherin cytosolic tail that is in contact with the ARM region. PMID:23083718

The Use of Climate Projections in the Modelling of Bud Burst

NASA Astrophysics Data System (ADS)

O'Neill, Bridget F.; Caffara, Amelia; Gleeson, Emily; Semmler, Tido; McGrath, Ray; Donnelly, Alison

2010-05-01

Recent changes in global climate, such as increasing temperature, have had notable effects on the phenology (timing of biological events) of plants. The effects are variable across habitats and between species, but increasing temperatures have been shown to advance certain key phenophases of trees, such as bud burst (beginning of leaf unfolding). This project considered climate change impacts on phenology of plants at a local scale in Ireland. The output from the ENSEMBLES climate simulations were down-scaled to Ireland and utilised by a phenological model to project changes over the next 50-100 years. This project helps to showcase the potential use of climate simulations in phenological research.

Application of plant growth regulators mitigates chlorotic foliar injury by the black pecan aphid (Hemiptera: Aphididae).

PubMed

Cottrell, Ted E; Wood, Bruce W; Ni, Xinzhi

2010-11-01

Black pecan aphid, Melanocallis caryaefoliae (Davis) (Hemiptera: Aphididae), feeding elicits localized chlorotic injury to pecan foliage [Carya illinoinensis (Wangenh.) K Koch] and apparent acceleration of leaf senescence and defoliation. The ability of certain plant growth regulators (PGRs) (forchlorfenuron, gibberellic acid and aviglycine) to prevent M. caryaefoliae from triggering pecan leaf chlorosis and senescence-like processes was evaluated on two dates in both 2006 and 2007. Treatments were applied to orchard foliage and used in laboratory leaf-disc bioassays to assess possible reduction in aphid-elicited chlorosis and concomitant effects on aphid mortality and development. Foliage pretreated with forchlorfenuron + gibberellic acid prior to being challenged with aphids resulted in significantly less aphid-elicited chlorosis than did control or aviglycine-treated leaf discs. No PGR affected aphid mortality; however, development time was increased by forchlorfenuron + gibberellic acid in 2006 and by aviglycine + gibberellic acid on one date in 2007. Certain PGRs possess the potential for usage on pecan to protect foliar canopies from M. caryaefoliae via changes in the susceptibility of the host leaf to senescence-like factors being introduced by feeding aphids. This protective effect on host foliage and the associated suppressive effect on development of feeding aphids might also be relevant to pest management programs on other aphid-crop systems in which aphid-elicited chlorosis and senescence-like processes can limit profitability. Published 2010 by John Wiley & Sons, Ltd.

Small heat-shock proteins and leaf cooling capacity account for the unusual heat tolerance of the central spike leaves in Agave tequilana var. Weber.

PubMed

Luján, Rosario; Lledías, Fernando; Martínez, Luz María; Barreto, Rita; Cassab, Gladys I; Nieto-Sotelo, Jorge

2009-12-01

Agaves are perennial crassulacean acid metabolism (CAM) plants distributed in tropical and subtropical arid environments, features that are attractive for studying the heat-shock response. In agaves, the stress response can be analysed easily during leaf development, as they form a spirally shaped rosette, having the meristem surrounded by folded leaves in the centre (spike) and the unfolded and more mature leaves in the periphery. Here, we report that the spike of Agave tequilana is the most thermotolerant part of the rosette withstanding shocks of up to 55 degrees C. This finding was inconsistent with the patterns of heat-shock protein (Hsp) gene expression, as maximal accumulation of Hsp transcripts was at 44 degrees C in all sectors (spike, inner, middle and outer). However, levels of small HSP (sHSP)-CI and sHSP-CII proteins were conspicuously higher in spike leaves at all temperatures correlating with their thermotolerance. In addition, spike leaves showed a higher stomatal density and abated more efficiently their temperature several degrees below that of air. We propose that the greater capacity for leaf cooling during the day in response to heat stress, and the elevated levels of sHSPs, constitute part of a set of strategies that protect the SAM and folded leaves of A. tequilana from high temperatures.

Leaf Pressure Volume Data in Caxiuana and Tapajos National Forest, Para, Brazil (2011)

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

Powell, Thomas; Moorcroft, Paul

Pressure volume curve measurements on leaves of canopy trees from the from the Caxiuana and Tapajos National Forests, Para, Brazil. Tapajos samples were harvested from the km 67 forested area, which is adjacent to the decommissioned throughfall exclusion drought experimental plot. Caxiuana samples were harvested from trees growing in the throughfall exclusion plots. Data were collected in 2011. Dataset includes: date of measurement, site ID, plot ID, tree ID (species, tree tag #), leaf area, fresh weight, relative weight, leaf water potential, and leaf water loss. P-V curve parameters (turgor loss point, osmotic potential, and bulk modulus of elasticity) canmore » be found in Powell et al. (2017) Differences in xylem cavitation resistance and leaf hydraulic traits explain differences in drought tolerance among mature Amazon rainforest trees. Global Change Biology.« less

Effects of management practices on reflectance of spring wheat canopies. [Williston, North Dakota Agricultural Experiment Station

NASA Technical Reports Server (NTRS)

Daughtry, C. S. T.; Bauer, M. E.; Crecelius, D. W.; Hixson, M. M. (Principal Investigator)

1980-01-01

The effects of available soil moisture, planting date, nitrogen fertilization, and cultivar on reflectance of spring wheat (Triticum aestivum L.) canopies were investigated. Spectral measurements were acquired on eight dates throughout the growing season, along with measurements of crop maturity stage, leaf area index, biomass, plant height, percent soil cover, and soil moisture. Planting date and available soil moisture were the primary agronomic factors which affected reflectance of spring wheat canopies from tillering to maturity. Comparisons of treatments indicated that during the seedling and tillering stages planting date was associated with 36 percent and 85 percent of variation in red and near infrared reflectances, respectively. As the wheat headed and matured, less of the variation in reflectance was associated with planting date and more with available soil moisture. By mid July, soil moisture accounted for 73 percent and 69 percent of the variation in reflectance in red and near infrared bands, respectively. Differences in spectral reflectance among treatments were attributed to changes in leaf area index, biomass, and percent soil cover. Cultivar and N fertilization rate were associated with very little of the variation in the reflectance of these canopies.

The spatial pattern of leaf phenology and its response to climate change in China.

PubMed

Dai, Junhu; Wang, Huanjiong; Ge, Quansheng

2014-05-01

Leaf phenology has been shown to be one of the most important indicators of the effects of climate change on biological systems. Few such studies have, however, been published detailing the relationship between phenology and climate change in Asian contexts. With the aim of quantifying species' phenological responsiveness to temperature and deepening understandings of spatial patterns of phenological and climate change in China, this study analyzes the first leaf date (FLD) and the leaf coloring date (LCD) from datasets of four woody plant species, Robinia pseudoacacia, Ulmus pumila, Salix babylonica, and Melia azedarach, collected from 1963 to 2009 at 47 Chinese Phenological Observation Network (CPON) stations spread across China (from 21° to 50° N). The results of this study show that changes in temperatures in the range of 39-43 days preceding the date of FLD of these plants affected annual variations in FLD, while annual variations in temperature in the range of 71-85 days preceding LCD of these plants affected the date of LCD. Average temperature sensitivity of FLD and LCD for these plants was -3.93 to 3.30 days °C(-1) and 2.11 to 4.43 days °C⁻¹, respectively. Temperature sensitivity of FLD was found to be stronger at lower latitudes or altitude as well as in more continental climates, while the response of LCD showed no consistent pattern. Within the context of significant warming across China during the study period, FLD was found to have advanced by 5.44 days from 1960 to 2009; over the same period, LCD was found to have been delayed by 4.56 days. These findings indicate that the length of the growing season of the four plant species studied was extended by a total of 10.00 days from 1960 to 2009. They also indicate that phenological response to climate is highly heterogeneous spatially.

Field-acclimated Gossypium hirsutum cultivars exhibit genotypic and seasonal differences in photosystem II thermostability.

PubMed

Snider, John L; Oosterhuis, Derrick M; Collins, Guy D; Pilon, Cristiane; Fitzsimons, Toby R

2013-03-15

Previous investigations have demonstrated that photosystem II (PSII) thermostability acclimates to prior exposure to heat and drought, but contrasting results have been reported for cotton (Gossypium hirsutum). We hypothesized that PSII thermotolerance in G. hirsutum would acclimate to environmental conditions during the growing season and that there would be differences in PSII thermotolerance between commercially-available U.S. cultivars. To this end, three cotton cultivars were grown under dryland conditions in Tifton Georgia, and two under irrigated conditions in Marianna Arkansas. At Tifton, measurements included PSII thermotolerance (T15, the temperature causing a 15% decline in maximum quantum yield), leaf temperatures, air temperatures, midday (1200 to 1400h) leaf water potentials (ΨMD), leaf-air vapor pressure deficit (VPD), actual quantum yield (ΦPSII) and electron transport rate through PSII (ETR) on three sample dates. At Marianna, T15 was measured on two sample dates. Optimal air and leaf temperatures were observed on all sample dates in Tifton, but PSII thermotolerance increased with water deficit conditions (ΨMD=-3.1MPa), and ETR was either unaffected or increased under water-stress. Additionally, T15 for PHY 499 was ∼5°C higher than for the other cultivars examined (DP 0912 and DP 1050). The Marianna site experienced more extreme high temperature conditions (20-30 days Tmax≥35°C), and showed an increase in T15 with higher average Tmax. When average T15 values for each location and sample date were plotted versus average daily Tmax, strong, positive relationships (r(2) from .954 to .714) were observed between Tmax and T15. For all locations T15 was substantially higher than actual field temperature conditions. We conclude that PSII thermostability in G. hirsutum acclimates to pre-existing environmental conditions; PSII is extremely tolerant to high temperature and water-deficit stress; and differences in PSII thermotolerance exist between commercially-available cultivars. Copyright © 2012 Elsevier GmbH. All rights reserved.

Patterns in leaf morphological traits of Chinese woody plants and the application for paleoclimate reconstruction

NASA Astrophysics Data System (ADS)

Li, Yaoqi; Wang, Zhiheng

2017-04-01

Leaf morphological traits (LMTs) directly influence carbon-uptake and water-loss of plants in different habitats, and hence can be sensitive indicators of plant interaction with climate. The relationships between community-aggregated LMTs and their surrounding climate have been used to reconstruct paleoclimate. However, the uncertainties in its application remain poorly explored. Using distribution maps and LMTs data (leaf margin states, leaf length, leaf width, and length-width product/ratio) of 10480 Chinese woody dicots and dated family-level phylogenies, we demonstrated the variations of LMTs in geographical patterns, and analyzed their relationships with climate across different life-forms (evergreen and deciduous; trees, shrubs and lianas) and species quartiles with different family-ages. Results showed that from southern to northern China, leaves became shorter and narrower, while leaf length-width ratio increased and toothed-margin percentage decreased. Our results revealed great uncertainties in leaf margin-temperature relationships induced by life-form, precipitation and evolutionary history, and suggested that the widely-used method, leaf margin analysis, should be applied cautiously on paleotemperature reconstruction. Differently, mean leaf size responded tightly to spatial variations in annual evapotranspiration (AET) and primary productivity (GPP and NPP), and these relationships remained constant across different life-forms and evolutionary history, suggesting that leaf size could be a useful surrogate for paleo primary productivity.

Partial unfolding and refolding for structure refinement: A unified approach of geometric simulations and molecular dynamics.

PubMed

Kumar, Avishek; Campitelli, Paul; Thorpe, M F; Ozkan, S Banu

2015-12-01

The most successful protein structure prediction methods to date have been template-based modeling (TBM) or homology modeling, which predicts protein structure based on experimental structures. These high accuracy predictions sometimes retain structural errors due to incorrect templates or a lack of accurate templates in the case of low sequence similarity, making these structures inadequate in drug-design studies or molecular dynamics simulations. We have developed a new physics based approach to the protein refinement problem by mimicking the mechanism of chaperons that rehabilitate misfolded proteins. The template structure is unfolded by selectively (targeted) pulling on different portions of the protein using the geometric based technique FRODA, and then refolded using hierarchically restrained replica exchange molecular dynamics simulations (hr-REMD). FRODA unfolding is used to create a diverse set of topologies for surveying near native-like structures from a template and to provide a set of persistent contacts to be employed during re-folding. We have tested our approach on 13 previous CASP targets and observed that this method of folding an ensemble of partially unfolded structures, through the hierarchical addition of contact restraints (that is, first local and then nonlocal interactions), leads to a refolding of the structure along with refinement in most cases (12/13). Although this approach yields refined models through advancement in sampling, the task of blind selection of the best refined models still needs to be solved. Overall, the method can be useful for improved sampling for low resolution models where certain of the portions of the structure are incorrectly modeled. © 2015 Wiley Periodicals, Inc.

Large seasonal swings in leaf area of Amazon rainforests

PubMed Central

Myneni, Ranga B.; Yang, Wenze; Nemani, Ramakrishna R.; Huete, Alfredo R.; Dickinson, Robert E.; Knyazikhin, Yuri; Didan, Kamel; Fu, Rong; Negrón Juárez, Robinson I.; Saatchi, Sasan S.; Hashimoto, Hirofumi; Ichii, Kazuhito; Shabanov, Nikolay V.; Tan, Bin; Ratana, Piyachat; Privette, Jeffrey L.; Morisette, Jeffrey T.; Vermote, Eric F.; Roy, David P.; Wolfe, Robert E.; Friedl, Mark A.; Running, Steven W.; Votava, Petr; El-Saleous, Nazmi; Devadiga, Sadashiva; Su, Yin; Salomonson, Vincent V.

2007-01-01

Despite early speculation to the contrary, all tropical forests studied to date display seasonal variations in the presence of new leaves, flowers, and fruits. Past studies were focused on the timing of phenological events and their cues but not on the accompanying changes in leaf area that regulate vegetation–atmosphere exchanges of energy, momentum, and mass. Here we report, from analysis of 5 years of recent satellite data, seasonal swings in green leaf area of ≈25% in a majority of the Amazon rainforests. This seasonal cycle is timed to the seasonality of solar radiation in a manner that is suggestive of anticipatory and opportunistic patterns of net leaf flushing during the early to mid part of the light-rich dry season and net leaf abscission during the cloudy wet season. These seasonal swings in leaf area may be critical to initiation of the transition from dry to wet season, seasonal carbon balance between photosynthetic gains and respiratory losses, and litterfall nutrient cycling in moist tropical forests. PMID:17360360

Tree litter and forest understorey vegetation: a conceptual framework to understand the effects of tree litter on a perennial geophyte, Anemone nemorosa.

PubMed

Baltzinger, Marie; Archaux, Frédéric; Dumas, Yann

2012-05-01

Litter is a key factor in structuring plant populations, through positive or negative interactions. The litter layer forms a mechanical barrier that is often strongly selective against individuals lacking hypocotyle plasticity. Litter composition also interacts with plant growth by providing beneficial nutrients or, inversely, by allowing harmful allelopathic leaching. As conspicuous litter fall accumulation is often observed under deciduous forests, interactions between tree litter and understorey plant populations are worthy of study. In a 1-year ex-situ experiment, the effects of tree litter on the growth of Anemone nemorosa, a small perennial forest geophyte, were investigated. Three 'litter quantity' treatments were defined, representative of forest floor litter (199, 356·5 and 514 g m(-2)), which were crossed with five 'litter composition' treatments (Quercus petraea, Fagus sylvatica, Carpinus betulus, Q. petraea + F. sylvatica and Q. petraea + C. betulus), plus a no-litter control. Path analysis was then used to investigate the pathways linking litter characteristics and components of adult plant growth. As expected, the heavier the litter, the longer the petiole; rhizome growth, however, was not depreciated by the litter-induced petiole lengthening. Both rhizome mass increment and number of initiated buds marginally increased with the amount of litter. Rhizome mass increment was in fact determined primarily by leaf area and leaf life span, neither of which was unequivocally correlated with any litter characteristics. However, the presence of litter significantly increased leafing success: following a late frost event, control rhizomes growing in the absence of litter experienced higher leaf mortality before leaf unfolding. The study questions the role of litter as a physical or chemical barrier to ground vegetation; to better understand this role, there is a need for ex-situ, longer-term experiments coupled with in-situ observations in the forest.

Tree litter and forest understorey vegetation: a conceptual framework to understand the effects of tree litter on a perennial geophyte, Anemone nemorosa

PubMed Central

Baltzinger, Marie; Archaux, Frédéric; Dumas, Yann

2012-01-01

Background and Aims Litter is a key factor in structuring plant populations, through positive or negative interactions. The litter layer forms a mechanical barrier that is often strongly selective against individuals lacking hypocotyle plasticity. Litter composition also interacts with plant growth by providing beneficial nutrients or, inversely, by allowing harmful allelopathic leaching. As conspicuous litter fall accumulation is often observed under deciduous forests, interactions between tree litter and understorey plant populations are worthy of study. Methods In a 1-year ex-situ experiment, the effects of tree litter on the growth of Anemone nemorosa, a small perennial forest geophyte, were investigated. Three ‘litter quantity’ treatments were defined, representative of forest floor litter (199, 356·5 and 514 g m−2), which were crossed with five ‘litter composition’ treatments (Quercus petraea, Fagus sylvatica, Carpinus betulus, Q. petraea + F. sylvatica and Q. petraea + C. betulus), plus a no-litter control. Path analysis was then used to investigate the pathways linking litter characteristics and components of adult plant growth. Key Results As expected, the heavier the litter, the longer the petiole; rhizome growth, however, was not depreciated by the litter-induced petiole lengthening. Both rhizome mass increment and number of initiated buds marginally increased with the amount of litter. Rhizome mass increment was in fact determined primarily by leaf area and leaf life span, neither of which was unequivocally correlated with any litter characteristics. However, the presence of litter significantly increased leafing success: following a late frost event, control rhizomes growing in the absence of litter experienced higher leaf mortality before leaf unfolding. Conclusions The study questions the role of litter as a physical or chemical barrier to ground vegetation; to better understand this role, there is a need for ex-situ, longer-term experiments coupled with in-situ observations in the forest. PMID:22419760

An artificial solar spectrum substantially alters plant development compared with usual climate room irradiance spectra.

PubMed

Hogewoning, Sander W; Douwstra, Peter; Trouwborst, Govert; van Ieperen, Wim; Harbinson, Jeremy

2010-03-01

Plant responses to the light spectrum under which plants are grown affect their developmental characteristics in a complicated manner. Lamps widely used to provide growth irradiance emit spectra which are very different from natural daylight spectra. Whereas specific responses of plants to a spectrum differing from natural daylight may sometimes be predictable, the overall plant response is generally difficult to predict due to the complicated interaction of the many different responses. So far studies on plant responses to spectra either use no daylight control or, if a natural daylight control is used, it will fluctuate in intensity and spectrum. An artificial solar (AS) spectrum which closely resembles a sunlight spectrum has been engineered, and growth, morphogenesis, and photosynthetic characteristics of cucumber plants grown for 13 d under this spectrum have been compared with their performance under fluorescent tubes (FTs) and a high pressure sodium lamp (HPS). The total dry weight of the AS-grown plants was 2.3 and 1.6 times greater than that of the FT and HPS plants, respectively, and the height of the AS plants was 4-5 times greater. This striking difference appeared to be related to a more efficient light interception by the AS plants, characterized by longer petioles, a greater leaf unfolding rate, and a lower investment in leaf mass relative to leaf area. Photosynthesis per leaf area was not greater for the AS plants. The extreme differences in plant response to the AS spectrum compared with the widely used protected cultivation light sources tested highlights the importance of a more natural spectrum, such as the AS spectrum, if the aim is to produce plants representative of field conditions.

Temporal variation in epidermal flavonoids due to altered solar UV radiation is moderated by the leaf position in Betula pendula.

PubMed

Morales, Luis O; Tegelberg, Riitta; Brosché, Mikael; Lindfors, Anders; Siipola, Sari; Aphalo, Pedro J

2011-11-01

The physiological mechanisms controlling plant responses to dynamic changes in ambient solar ultraviolet (UV) radiation are not fully understood: this information is important to further comprehend plant adaptation to their natural habitats. We used the fluorimeter Dualex to estimate in vivo the epidermal flavonoid contents by measuring epidermal UV absorbance (A(375) ) in Betula pendula Roth (silver birch) leaves of different ages under altered UV. Seedlings were grown in a greenhouse for 15 days without UV and transferred outdoors under three UV treatments (UV-0, UV-A and UV-A+B) created by three types of plastic film. After 7 and 13 days, Dualex measurements were taken at adaxial and abaxial epidermis of the first three leaves (L1, L2 and L3) of the seedlings. After 14 days, some of the seedlings were reciprocally swapped amongst the treatments to study the accumulation of epidermal flavonoids in the youngest unfolded leaves (L3) during leaf expansion under changing solar UV environments. A(375) of the leaves responded differently to the UV treatment depending on their position. UV-B increased the A(375) in the leaves independently of leaf position. L3 quickly adjusted A(375) in their epidermis according to the UV they received and these adjustments were affected by previous UV exposure. The initial absence of UV-A+B or UV-A, followed by exposure to UV-A+B, particularly enhanced leaf A(375) . Silver birch leaves modulate their protective pigments in response to changes in the UV environment during their expansion, and their previous UV exposure history affects the epidermal-absorbance achieved during later UV exposure. Copyright © Physiologia Plantarum 2011.

Dynamically downscaling predictions for deciduous tree leaf emergence in California under current and future climate.

PubMed

Medvigy, David; Kim, Seung Hee; Kim, Jinwon; Kafatos, Menas C

2016-07-01

Models that predict the timing of deciduous tree leaf emergence are typically very sensitive to temperature. However, many temperature data products, including those from climate models, have been developed at a very coarse spatial resolution. Such coarse-resolution temperature products can lead to highly biased predictions of leaf emergence. This study investigates how dynamical downscaling of climate models impacts simulations of deciduous tree leaf emergence in California. Models for leaf emergence are forced with temperatures simulated by a general circulation model (GCM) at ~200-km resolution for 1981-2000 and 2031-2050 conditions. GCM simulations are then dynamically downscaled to 32- and 8-km resolution, and leaf emergence is again simulated. For 1981-2000, the regional average leaf emergence date is 30.8 days earlier in 32-km simulations than in ~200-km simulations. Differences between the 32 and 8 km simulations are small and mostly local. The impact of downscaling from 200 to 8 km is ~15 % smaller in 2031-2050 than in 1981-2000, indicating that the impacts of downscaling are unlikely to be stationary.

Photosynthetic capacity regulation is uncoupled from nutrient limitation

NASA Astrophysics Data System (ADS)

Smith, N. G.; Keenan, T. F.; Prentice, I. C.; Wang, H.

2017-12-01

Ecosystem and Earth system models need information on leaf-level photosynthetic capacity, but to date typically rely on empirical estimates and an assumed dependence on nitrogen supply. Recent evidence suggests that leaf nitrogen is actively controlled though plant responses to photosynthetic demand. Here, we propose and test a theory of demand-driven coordination of photosynthetic processes, and use it to assess the relative roles of nutrient supply and photosynthetic demand. The theory captured 63% of observed variability in a global dataset of Rubisco carboxylation capacity (Vcmax; 3,939 values at 219 sites), suggesting that environmentally regulated biophysical costs and light availability are the first-order drivers of photosynthetic capacity. Leaf nitrogen, on the other hand, was a weak secondary driver of Vcmax, explaining less than 6% of additional observed variability. We conclude that leaf nutrient allocation is primarily driven by demand. Our theory offers a simple, robust strategy for dynamically predicting leaf-level photosynthetic capacity in global models.

Retrieval of Spatio-temporal Evaporation by Integrating Landsat OLI Optical and Thermal Data

NASA Astrophysics Data System (ADS)

Wandera, L. N.; Tol, C. V. D.; Mallick, K.; Bayat, B.; Verbeiren, B.; van Griensven, A.; Verhoef, W.; Suliga, J.; Barrios, J. M.; Chormański, J.; Kleniewska, M.

2017-12-01

Soil-Vegetation-Atmosphere (SVAT) Transfer Models are capable of providing continuous predictions of evapotranspiration (ET). However, providing these models with reliable spatio-temporal information of vegetation and soil properties remains challenging. Thus, combining optical and thermal satellite information might assists to overcome this challenge when using SVAT models. In this study, using a radiative transfer model of solar and sky radiation (RTMo), we simulate Landsat 8 reflectance bands (2-7). We then apply a numerical optimization approach to invert the model and retrieve the corresponding canopy attributes leaf chlorophyll content (Cab), leaf water content (Cw), leaf dry matter content (Cdm), leaf brown material (Cs), Leaf Area Index (LAI) and the leaf angle distribution function in the canopy at overpass time. The retrievals are then directly used as inputs into our SVAT model of choice, Soil Canopy Observations of Photochemistry and Energy Fluxes (SCOPE). Using a model for transfer of thermal radiation emitted by vegetation and soil (RTMt), we proceed to simulate Landsat radiance for the corresponding reflectance data using a lookup table (LUT). These variables were then used to develop a crop factor (Kc) map. A reference ET was generated and applied to the Kc map to obtain actual ET. We proceeded to interpolate the ET between the image acquisition dates to have a complete time series. The retrieval maps for the specific variables captured seasonal variability patterns for the respective variables. The generated KC map showed similar trend with the LAI maps. There was an underestimation of actual ET when the simulation was not constrained to the thermal information. The interpolation of ET between acquisition image dates reflected the seasonal trends. Key Word: SVAT, optical, thermal, remote sensing, evapotranspiration

Analysis on Difference of Forest Phenology Extracted from EVI and LAI Based on PhenoCams

NASA Astrophysics Data System (ADS)

Wang, C.; Jing, L.; Qinhuo, L.

2017-12-01

Land surface phenology can make up for the deficiency of field observation with advantages of capturing the continuous expression of phenology on a large scale. However, there are some variability in phenological metrics derived from different satellite time-series data of vegetation parameters. This paper aims at assessing the difference of phenology information extracted from EVI and LAI time series. To achieve this, some web-camera sites were selected to analyze the characteristics between MODIS-EVI and MODIS-LAI time series from 2010 to 2014 for different forest types, including evergreen coniferous forest, evergreen broadleaf forest, deciduous coniferous forest and deciduous broadleaf forest. At the same time, satellite-based phenological metrics were extracted by the Logistics algorithm and compared with camera-based phenological metrics. Results show that the SOS and EOS that are extracted from LAI are close to bud burst and leaf defoliation respectively, while the SOS and EOS that are extracted from EVI is close to leaf unfolding and leaf coloring respectively. Thus the SOS that is extracted from LAI is earlier than that from EVI, while the EOS that is extracted from LAI is later than that from EVI at deciduous forest sites. Although the seasonal variation characteristics of evergreen forests are not apparent, significant discrepancies exist in LAI time series and EVI time series. In addition, Satellite- and camera-based phenological metrics agree well generally, but EVI has higher correlation with the camera-based canopy greenness (green chromatic coordinate, gcc) than LAI.

Trends and Variability in Temperature Sensitivity of Lilac Flowering Phenology

NASA Astrophysics Data System (ADS)

Wang, Huanjiong; Dai, Junhu; Rutishauser, This; Gonsamo, Alemu; Wu, Chaoyang; Ge, Quansheng

2018-03-01

The responses of plant phenology to temperature variability have many consequences for ecological processes, agriculture, forestry, and human health. Temperature sensitivity (ST) of phenology could measure how and to what degree plant could phenologically track climate change. The long-term trends and spatial patterns in ST have been well studied for vegetative phenology such as leaf unfolding, but trends to be expected for reproductive phenology in the future remain unknown. Here we investigate trends and factors driving the temporal variation of ST of first bloom date (FBD). Using the long-term FBD records during 1963-2013 for common lilac (Syringa vulgaris) from 613 stations in Europe, we compared changes in ST from the beginning to the end of the study period. The Spearman partial correlations were used to assess the importance of four influencing factors. The results showed that the temporal changes in ST of FBD varied considerably among time scales. Mean ST decreased significantly by 0.92 days °C-1 from 1963-1972 to 2004-2013 (P < 0.01), but remained stable from 1963-1987 to 1989-2013. The strength of FBD and temperature relationship, the spring temperature variance, and winter chill all impact ST in an expected way at most stations. No consistent responses of ST on photoperiod were found. Our results imply that the trends and variability in ST of flowering phenology are driving by multiple factors and impacted by time scales. Continued efforts are still needed to further examine the flowering-temperature relationship for other plant species in other climates and environments using similar methods to our study.

Role of Aquaporins in a Composite Model of Water Transport in the Leaf.

PubMed

Yaaran, Adi; Moshelion, Menachem

2016-06-30

Water-transport pathways through the leaf are complex and include several checkpoints. Some of these checkpoints exhibit dynamic behavior that may be regulated by aquaporins (AQPs). To date, neither the relative weight of the different water pathways nor their molecular mechanisms are well understood. Here, we have collected evidence to support a putative composite model of water pathways in the leaf and the distribution of water across those pathways. We describe how water moves along a single transcellular path through the parenchyma and continues toward the mesophyll and stomata along transcellular, symplastic and apoplastic paths. We present evidence that points to a role for AQPs in regulating the relative weight of each path in the overall leaf water-transport system and the movement of water between these paths as a result of the integration of multiple signals, including transpiration demand, water potential and turgor. We also present a new theory, the hydraulic fuse theory, to explain effects of the leaf turgor-loss-point on water paths alternation and the subsequent reduction in leaf hydraulic conductivity. An improved understating of leaf water-balance management may lead to the development of crops that use water more efficiently, and responds better to environmental changes.

An extended PROSPECT: Advance in the leaf optical properties model separating total chlorophylls into chlorophyll a and b.

PubMed

Zhang, Yao; Huang, Jingfeng; Wang, Fumin; Blackburn, George Alan; Zhang, Hankui K; Wang, Xiuzhen; Wei, Chuanwen; Zhang, Kangyu; Wei, Chen

2017-07-25

The PROSPECT leaf optical model has, to date, well-separated the effects of total chlorophyll and carotenoids on leaf reflectance and transmittance in the 400-800 nm. Considering variations in chlorophyll a:b ratio with leaf age and physiological stress, a further separation of total plant-based chlorophylls into chlorophyll a and chlorophyll b is necessary for advanced monitoring of plant growth. In this study, we present an extended version of PROSPECT model (hereafter referred to as PROSPECT-MP) that can combine the effects of chlorophyll a, chlorophyll b and carotenoids on leaf directional hemispherical reflectance and transmittance (DHR and DHT) in the 400-800 nm. The LOPEX93 dataset was used to evaluate the capabilities of PROSPECT-MP for spectra modelling and pigment retrieval. The results show that PROSPECT-MP can both simultaneously retrieve leaf chlorophyll a and b, and also performs better than PROSPECT-5 in retrieving carotenoids concentrations. As for the simulation of DHR and DHT, the performances of PROSPECT-MP are similar to that of PROSPECT-5. This study demonstrates the potential of PROSPECT-MP for improving capabilities of remote sensing of leaf photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoids) and for providing a framework for future refinements in the modelling of leaf optical properties.

Study of multiple unfolding trajectories and unfolded states of the protein GB1 under the physical property space.

PubMed

Wang, Jihua; Zhao, Liling; Dou, Xianghua; Zhang, Zhiyong

2008-06-01

Forty nine molecular dynamics simulations of unfolding trajectories of the segment B1 of streptococcal protein G (GB1) provide a direct demonstration of the diversity of unfolding pathway and give a statistically utmost unfolding pathway under the physical property space. Twelve physical properties of the protein were chosen to construct a 12-dimensional property space. Then the 12-dimensional property space was reduced to a 3-dimensional principle component property space. Under the property space, the multiple unfolding trajectories look like "trees", which have some common characters. The "root of the tree" corresponds to the native state, the "bole" homologizes the partially unfolded conformations, and the "crown" is in correspondence to the unfolded state. These unfolding trajectories can be divided into three types. The first one has the characters of straight "bole" and "crown" corresponding to a fast two-state unfolding pathway of GB1. The second one has the character of "the standstill in the middle tree bole", which may correspond to a three-state unfolding pathway. The third one has the character of "the circuitous bole" corresponding to a slow two-state unfolding pathway. The fast two-state unfolding pathway is a statistically utmost unfolding pathway or preferred pathway of GB1, which occupies 53% of 49 unfolding trajectories. In the property space all the unfolding trajectories construct a thermal unfolding pathway ensemble of GB1. The unfolding pathway ensemble resembles a funnel that is gradually emanative from the native state ensemble to the unfolded state ensemble. In the property space, the thermal unfolded state distribution looks like electronic cloud in quantum mechanics. The unfolded states of the independent unfolding simulation trajectories have substantial overlaps, indicating that the thermal unfolded states are confined by the physical property values, and the number of protein unfolded state are much less than that was believed before.

Contribution of flavonoids to the overall radical scavenging activity of olive (Olea europaea L.) leaf polar extracts.

PubMed

Goulas, Vlassios; Papoti, Vassiliki T; Exarchou, Vassiliki; Tsimidou, Maria Z; Gerothanassis, Ioannis P

2010-03-24

The contribution of flavonoids to the overall radical scavenging activity of olive leaf polar extracts, known to be good sources of oleuropein related compounds, was examined. Off line and on line HPLC-DPPH(*) assays were employed, whereas flavonoid content was estimated colorimetrically. Individual flavonoid composition was first assessed by RP-HPLC coupled with diode array and fluorescence detectors and verified by LC-MS detection system. Olive leaf was found a robust source of flavonoids regardless sampling parameters (olive cultivar, leaf age or sampling date). Total flavonoids accounted for the 13-27% of the total radical scavenging activity assessed using the on line protocol. Luteolin 7-O-glucoside was one of the dominant scavengers (8-25%). Taking into consideration frequency of appearance the contribution of luteolin (3-13%) was considered important, too. Our findings support that olive leaf, except for oleuropein and related compounds, is also a stable source of bioactive flavonoids.

Substantial variation in leaf senescence times among 1360 temperate woody plant species: implications for phenology and ecosystem processes

PubMed Central

Panchen, Zoe A.; Primack, Richard B.; Gallinat, Amanda S.; Nordt, Birgit; Stevens, Albert-Dieter; Du, Yanjun; Fahey, Robert

2015-01-01

Background and Aims Autumn leaf senescence marks the end of the growing season in temperate ecosystems. Its timing influences a number of ecosystem processes, including carbon, water and nutrient cycling. Climate change is altering leaf senescence phenology and, as those changes continue, it will affect individual woody plants, species and ecosystems. In contrast to spring leaf out times, however, leaf senescence times remain relatively understudied. Variation in the phenology of leaf senescence among species and locations is still poorly understood. Methods Leaf senescence phenology of 1360 deciduous plant species at six temperate botanical gardens in Asia, North America and Europe was recorded in 2012 and 2013. This large data set was used to explore ecological and phylogenetic factors associated with variation in leaf senescence. Key Results Leaf senescence dates among species varied by 3 months on average across the six locations. Plant species tended to undergo leaf senescence in the same order in the autumns of both years at each location, but the order of senescence was only weakly correlated across sites. Leaf senescence times were not related to spring leaf out times, were not evolutionarily conserved and were only minimally influenced by growth habit, wood anatomy and percentage colour change or leaf drop. These weak patterns of leaf senescence timing contrast with much stronger leaf out patterns from a previous study. Conclusions The results suggest that, in contrast to the broader temperature effects that determine leaf out times, leaf senescence times are probably determined by a larger or different suite of local environmental effects, including temperature, soil moisture, frost and wind. Determining the importance of these factors for a wide range of species represents the next challenge for understanding how climate change is affecting the end of the growing season and associated ecosystem processes. PMID:25808654

Substantial variation in leaf senescence times among 1360 temperate woody plant species: implications for phenology and ecosystem processes.

PubMed

Panchen, Zoe A; Primack, Richard B; Gallinat, Amanda S; Nordt, Birgit; Stevens, Albert-Dieter; Du, Yanjun; Fahey, Robert

2015-11-01

Autumn leaf senescence marks the end of the growing season in temperate ecosystems. Its timing influences a number of ecosystem processes, including carbon, water and nutrient cycling. Climate change is altering leaf senescence phenology and, as those changes continue, it will affect individual woody plants, species and ecosystems. In contrast to spring leaf out times, however, leaf senescence times remain relatively understudied. Variation in the phenology of leaf senescence among species and locations is still poorly understood. Leaf senescence phenology of 1360 deciduous plant species at six temperate botanical gardens in Asia, North America and Europe was recorded in 2012 and 2013. This large data set was used to explore ecological and phylogenetic factors associated with variation in leaf senescence. Leaf senescence dates among species varied by 3 months on average across the six locations. Plant species tended to undergo leaf senescence in the same order in the autumns of both years at each location, but the order of senescence was only weakly correlated across sites. Leaf senescence times were not related to spring leaf out times, were not evolutionarily conserved and were only minimally influenced by growth habit, wood anatomy and percentage colour change or leaf drop. These weak patterns of leaf senescence timing contrast with much stronger leaf out patterns from a previous study. The results suggest that, in contrast to the broader temperature effects that determine leaf out times, leaf senescence times are probably determined by a larger or different suite of local environmental effects, including temperature, soil moisture, frost and wind. Determining the importance of these factors for a wide range of species represents the next challenge for understanding how climate change is affecting the end of the growing season and associated ecosystem processes. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Persistent negative temperature response of mesophyll conductance in red raspberry (Rubus idaeus L.) leaves under both high and low vapour pressure deficits: a role for abscisic acid?

PubMed

Qiu, Changpeng; Ethier, Gilbert; Pepin, Steeve; Dubé, Pascal; Desjardins, Yves; Gosselin, André

2017-09-01

The temperature dependence of mesophyll conductance (g m ) was measured in well-watered red raspberry (Rubus idaeus L.) plants acclimated to leaf-to-air vapour pressure deficit (VPDL) daytime differentials of contrasting amplitude, keeping a fixed diurnal leaf temperature (T leaf ) rise from 20 to 35 °C. Contrary to the great majority of g m temperature responses published to date, we found a pronounced reduction of g m with increasing T leaf irrespective of leaf chamber O 2 level and diurnal VPDL regime. Leaf hydraulic conductance was greatly enhanced during the warmer afternoon periods under both low (0.75 to 1.5 kPa) and high (0.75 to 3.5 kPa) diurnal VPDL regimes, unlike stomatal conductance (g s ), which decreased in the afternoon. Consequently, the leaf water status remained largely isohydric throughout the day, and therefore cannot be evoked to explain the diurnal decrease of g m . However, the concerted diurnal reductions of g m and g s were well correlated with increases in leaf abscisic acid (ABA) content, thus suggesting that ABA can induce a significant depression of g m under favourable leaf water status. Our results challenge the view that the temperature dependence of g m can be explained solely from dynamic leaf anatomical adjustments and/or from the known thermodynamic properties of aqueous solutions and lipid membranes.​. © 2017 John Wiley & Sons Ltd.

Development of a Model to Predict the Primary Infection Date of Bacterial Spot (Xanthomonas campestris pv. vesicatoria) on Hot Pepper.

PubMed

Kim, Ji-Hoon; Kang, Wee-Soo; Yun, Sung-Chul

2014-06-01

A population model of bacterial spot caused by Xanthomonas campestris pv. vesicatoria on hot pepper was developed to predict the primary disease infection date. The model estimated the pathogen population on the surface and within the leaf of the host based on the wetness period and temperature. For successful infection, at least 5,000 cells/ml of the bacterial population were required. Also, wind and rain were necessary according to regression analyses of the monitored data. Bacterial spot on the model is initiated when the pathogen population exceeds 10(15) cells/g within the leaf. The developed model was validated using 94 assessed samples from 2000 to 2007 obtained from monitored fields. Based on the validation study, the predicted initial infection dates varied based on the year rather than the location. Differences in initial infection dates between the model predictions and the monitored data in the field were minimal. For example, predicted infection dates for 7 locations were within the same month as the actual infection dates, 11 locations were within 1 month of the actual infection, and only 3 locations were more than 2 months apart from the actual infection. The predicted infection dates were mapped from 2009 to 2012; 2011 was the most severe year. Although the model was not sensitive enough to predict disease severity of less than 0.1% in the field, our model predicted bacterial spot severity of 1% or more. Therefore, this model can be applied in the field to determine when bacterial spot control is required.

Development of a Model to Predict the Primary Infection Date of Bacterial Spot (Xanthomonas campestris pv. vesicatoria) on Hot Pepper

PubMed Central

Kim, Ji-Hoon; Kang, Wee-Soo; Yun, Sung-Chul

2014-01-01

A population model of bacterial spot caused by Xanthomonas campestris pv. vesicatoria on hot pepper was developed to predict the primary disease infection date. The model estimated the pathogen population on the surface and within the leaf of the host based on the wetness period and temperature. For successful infection, at least 5,000 cells/ml of the bacterial population were required. Also, wind and rain were necessary according to regression analyses of the monitored data. Bacterial spot on the model is initiated when the pathogen population exceeds 1015 cells/g within the leaf. The developed model was validated using 94 assessed samples from 2000 to 2007 obtained from monitored fields. Based on the validation study, the predicted initial infection dates varied based on the year rather than the location. Differences in initial infection dates between the model predictions and the monitored data in the field were minimal. For example, predicted infection dates for 7 locations were within the same month as the actual infection dates, 11 locations were within 1 month of the actual infection, and only 3 locations were more than 2 months apart from the actual infection. The predicted infection dates were mapped from 2009 to 2012; 2011 was the most severe year. Although the model was not sensitive enough to predict disease severity of less than 0.1% in the field, our model predicted bacterial spot severity of 1% or more. Therefore, this model can be applied in the field to determine when bacterial spot control is required. PMID:25288995

Improving classification accuracy using multi-date IRS/LISS data and development of thermal stress index for Asiatic lion habitat

NASA Astrophysics Data System (ADS)

Gupta, Rajendra Kumar

The increase in lion and leopard population in the GIR wild life sanctuary and National Park (Gir Protected Area) demands periodic and precision monitoring of habitat at close intervals using space based remote sensing data. Besides characterizing the different forest classes, remote sensing needs to support for the assessment of thermal stress zones and identification of possible corridors for lion dispersion to new home ranges. The study focuses on assessing the thematic forest classification accuracies in percentage terms(CA) attainable using single date post-monsoon (CA=60, kappa = 0.514) as well as leaf shedding (CA=48.4, kappa = 0.372) season data in visible and Near-IR spectral bands of IRS/LISS-III at 23.5 m spatial resolution; and improvement of CA by using joint two date (multi-temporal) data sets (CA=87.2, kappa = 0.843) in the classification. The 188 m spatial resolution IRS/WiFS and 23.5 m spatial resolution LISS-III data were used to study the possible corridors for dispersion of Lions from GIR protected areas (PA). A relative thermal stress index (RTSI) for Gir PA has been developed using NOAA/ AVHRR data sets of post-monsoon, leaf shedded and summer seasons. The paper discusses the role of RTSI as a tool to work out forest management plans using leaf shedded season data to combat the thermal stress in the habitat, by identifying locations for artificial water holes during the ensuing summer season.

Identification of Candidate Genes Involved in the Salt Tolerance of Date Palm (Phoenix dactylifera L.) Based on a Yeast Functional Bioassay.

PubMed

Patankar, Himanshu V; Al-Harrasi, Ibtisam; Al-Yahyai, Rashid; Yaish, Mahmoud W

2018-06-01

Although date palm is a relatively salt-tolerant plant, the molecular basis of this tolerance is complex and poorly understood. Therefore, this study aimed to identify the genes involved in salinity tolerance using a basic yeast functional bioassay. To achieve this, a date palm cDNA library was overexpressed in Saccharomyces cerevisiae cells. The expression levels of selected genes that make yeast cells tolerant to salt were subsequently validated in the leaf and root tissues of date palm seedlings using a quantitative PCR method. About 6000 yeast transformant cells were replica printed and screened on a synthetic minimal medium containing 1.0 M of NaCl. The screening results showed the presence of 62 salt-tolerant transformant colonies. Sequence analysis of the recombinant yeast plasmids revealed the presence of a group of genes with potential salt-tolerance functions, such as aquaporins (PIP), serine/threonine protein kinases (STKs), ethylene-responsive transcription factor 1 (ERF1), and peroxidases (PRX). The expression pattern of the selected genes endorsed the hypothesis that these genes may be involved in salinity tolerance, as they showed a significant (p < 0.05) overexpression trend in both the leaf and root tissues in response to salinity. The genes identified in this project are suitable candidates for the further functional characterization of date palms.

Applying remote sensing measurements of phenology to southern California vegetation

NASA Astrophysics Data System (ADS)

Willis, K. S.; Gillespie, T. W.

2012-12-01

Monitoring vegetation phenology can be used to assess the impacts of climate change on a localized region. This study aims to determine the most applicable remote sensing method for monitoring phenological changes in the largest urban National Park in the US: the Santa Monica Mountains of southern California. This is achieved by comparing the Normalized Difference Vegetation Index (NDVI), considered applicable to Mediterranean-type ecosystems due to the low amount of greenness present in the vegetation, with relative spectral mixture analysis (RMSA). RMSA is a technique developed to measure temporal changes in green vegetation (GV), nonphotosynthetic vegetation plus litter (NPV), and snow cover designed for the south-central US. This study analyzes areas of natural vegetation in the Santa Monica Mountains using MODIS imagery by comparing GV and NPV indices derived from RMSA with the classic NDVI. The phenological transition dates of focus here include: (1) greenup, the date of onset of photosynthetic activity; (2) maturity, the date at which plant green leaf area is maximum; (3) senescence, the date at which photosynthetic activity and green leaf area begin to rapidly decrease; (4) dormancy, the date at which physiological activity becomes near zero. Overall, this study tests the application of RMSA to a new environment, compares these results to those derived from NDVI, and provides insight regarding the impacts of climate change on southern California phenological cycles.

Long-term temporal changes in central European tree phenology (1946-2010) confirm the recent extension of growing seasons.

PubMed

Kolářová, Eva; Nekovář, Jiří; Adamík, Peter

2014-10-01

One of the ways to assess the impacts of climate change on plants is analysing their long-term phenological data. We studied phenological records of 18 common tree species and their 8 phenological phases, spanning 65 years (1946-2010) and covering the area of the Czech Republic. For each species and phenophase, we assessed the changes in its annual means (for detecting shifts in the timing of the event) and standard deviations (for detecting changes in duration of the phenophases). The prevailing pattern across tree species was that since around the year 1976, there has been a consistent advancement of the onset of spring phenophases (leaf unfolding and flowering) and subsequent acceleration of fruit ripening, and a delay of autumn phenophases (leaf colouring and leaf falling). The most considerable shifts in the timing of spring phenophases were displayed by early-successional short-lived tree species. The most pronounced temporal shifts were found for the beginning of seed ripening in conifers with an advancement in this phenophase of up to 2.2 days year⁻¹ in Scots Pine (Pinus sylvestris). With regards to the change in duration of the phenophases, no consistent patterns were revealed. The growing season has extended on average by 23.8 days during the last 35 years. The most considerable prolongation was found in Pedunculate Oak (Quercus robur): 31.6 days (1976-2010). Extended growing season lengths do have the potential to increase growth and seed productivity, but unequal shifts among species might alter competitive relationships within ecosystems.

Long-term temporal changes in central European tree phenology (1946-2010) confirm the recent extension of growing seasons

NASA Astrophysics Data System (ADS)

Kolářová, Eva; Nekovář, Jiří; Adamík, Peter

2014-10-01

One of the ways to assess the impacts of climate change on plants is analysing their long-term phenological data. We studied phenological records of 18 common tree species and their 8 phenological phases, spanning 65 years (1946-2010) and covering the area of the Czech Republic. For each species and phenophase, we assessed the changes in its annual means (for detecting shifts in the timing of the event) and standard deviations (for detecting changes in duration of the phenophases). The prevailing pattern across tree species was that since around the year 1976, there has been a consistent advancement of the onset of spring phenophases (leaf unfolding and flowering) and subsequent acceleration of fruit ripening, and a delay of autumn phenophases (leaf colouring and leaf falling). The most considerable shifts in the timing of spring phenophases were displayed by early-successional short-lived tree species. The most pronounced temporal shifts were found for the beginning of seed ripening in conifers with an advancement in this phenophase of up to 2.2 days year-1 in Scots Pine ( Pinus sylvestris). With regards to the change in duration of the phenophases, no consistent patterns were revealed. The growing season has extended on average by 23.8 days during the last 35 years. The most considerable prolongation was found in Pedunculate Oak ( Quercus robur): 31.6 days (1976-2010). Extended growing season lengths do have the potential to increase growth and seed productivity, but unequal shifts among species might alter competitive relationships within ecosystems.

Comparing the intra-annual wood formation of three European species (Fagus sylvatica, Quercus petraea and Pinus sylvestris) as related to leaf phenology and non-structural carbohydrate dynamics.

PubMed

Michelot, Alice; Simard, Sonia; Rathgeber, Cyrille; Dufrêne, Eric; Damesin, Claire

2012-08-01

Monitoring cambial phenology and intra-annual growth dynamics is a useful approach for characterizing the tree growth response to climate change. However, there have been few reports concerning intra-annual wood formation in lowland temperate forests with high time resolution, especially for the comparison between deciduous and coniferous species. The main objective of this study was to determine how the timing, duration and rate of radial growth change between species as related to leaf phenology and the dynamics of non-structural carbohydrates (NSC) under the same climatic conditions. We studied two deciduous species, Fagus sylvatica L. and Quercus petraea (Matt.) Liebl., and an evergreen conifer, Pinus sylvestris L. During the 2009 growing season, we weekly monitored (i) the stem radial increment using dendrometers, (ii) the xylem growth using microcoring and (iii) the leaf phenology from direct observations of the tree crowns. The NSC content was also measured in the eight last rings of the stem cores in April, June and August 2009. The leaf phenology, NSC storage and intra-annual growth were clearly different between species, highlighting their contrasting carbon allocation. Beech growth began just after budburst, with a maximal growth rate when the leaves were mature and variations in the NSC content were low. Thus, beech radial growth seemed highly dependent on leaf photosynthesis. For oak, earlywood quickly developed before budburst, which probably led to the starch decrease quantified in the stem from April to June. For pine, growth began before the needles unfolding and the lack of NSC decrease during the growing season suggested that the substrates for radial growth were new assimilates of the needles from the previous year. Only for oak, the pattern determined from the intra-annual growth measured using microcoring differed from the pattern determined from dendrometer data. For all species, the ring width was significantly influenced by growth duration and not by growth rate, which differs from previous studies. The observed between-species difference at the intra-annual scale is key information for anticipating suitability of future species in temperate forests.

BOREAS TE-9 NSA Leaf Chlorophyll Density

NASA Technical Reports Server (NTRS)

Hall, Forrest G. (Editor); Curd, Shelaine (Editor); Margolis, Hank; Sy, Mikailou

2000-01-01

The BOREAS TE-9 team collected several data sets related to chemical and photosynthetic properties of leaves in boreal forest tree species. These data were collected to help provide an explanation of potential seasonal and spatial changes of leaf pigment properties in boreal forest species at the NSA. At different dates (FFC-Winter, FFC-Thaw, IFC-1, IFC-2, and IMC-3), foliage samples were collected from the upper third of the canopy for five NSA sites (YJP, OJP, OBS, UBS, and OA) near Thompson, Manitoba. Subsamples of 100 needles for black spruce, 20 needles for jack pine, and single leaf for trembling aspen were cut into pieces and immersed in a 20-mL DMF aliquot in a Nalgene test tube. The extracted foliage materials were then oven-dried at 68 C for 48 hours and weighed. Extracted leaf dry weight was converted to a total leaf area basis to express the chlorophyll content in mg/sq cm of total leaf area. The data are provided in tabular ASCII files. The data files are available on a CD-ROM (see document number 20010000884), or from the Oak Ridge National Laboratory (ORNL) Distributed Active Archive Center (DAAC).

Green leaf phenology at Landsat resolution: scaling from the plot to satellite

NASA Astrophysics Data System (ADS)

Fisher, J. I.; Mustard, J. F.; Vadeboncour, M.

2005-12-01

Despite the large number of in situ, plot-level phenological measurements and satellite-derived phenological studies, there has been little success to date in merging these records temporally or spatially. In particular, while most phenological patterns and trends derived from satellites appear realistic and coherent, they may not reflect spatial and temporal patterns at the plot level. An obvious explanation is the drastic scale difference from plot-level to most satellite observations. In this research, we bridge this scale gap through higher resolution satellite records (Landsat) and quantify the accuracy of satellite-derived metrics with direct field measurements. We compiled fifty-seven Landsat scenes from southern New England (P12 R51) from 1984 to 2002. Green vegetation areal abundance for each scene was derived from spectral mixture analysis and a single set of endmembers. The leaf area signal was fit with a logistic-growth simulating sigmoid curve to derive phenological markers (half-maximum leaf-onset and offset). Spring leaf-onset dates in homogenous stands of deciduous forests displayed significant and persistent local variability. The local variability was validated with multiple springtime ground observations (r2 = 0.91). The highest degree of verified small-scale variation occurred where contiguous forests displayed leaf-onset gradients of 10-14 days over short distances (<500 m). These dramatic gradients, of a similar magnitude to differences in leaf-onset from MD to MA, occur in of low-relief (<40 m) upland regions. The patterns suggest that microclimates resulting from springtime cold-air drainage may be influential in governing the start of leaf growth. These microclimates may be of crucial importance in interpreting in situ records and interpolating phenology from satellite data. Regional patterns from the Landsat analyses suggest topographic, coastal, and land-use controls on phenology. For example, our results indicate that deciduous forests in the Providence, RI metropolitan area leaf out 5-7 days earlier than comparable rural areas. In preliminary work, we validated the Landsat-derived metrics with similar analyses of MODIS and AVHRR, and demonstrate that aggregating diverse local phenologies into coarse grids may convolute interpretations. Despite these complications, the platform-independent curve-fit methodology may be extended across platforms and field data. The methodologically consistent approach, in tandem with Landsat data, allows us to effectively scale between plot and satellite phenological observations.

Cooperative unfolding of apolipoprotein A-1 induced by chemical denaturation.

PubMed

Eckhardt, D; Li-Blatter, X; Schönfeld, H-J; Heerklotz, H; Seelig, J

2018-05-25

Apolipoprotein A-1 (Apo A-1) plays an important role in lipid transfer and obesity. Chemical unfolding of α-helical Apo A-1 is induced with guanidineHCl and monitored with differential scanning calorimetry (DSC) and CD spectroscopy. The unfolding enthalpy and the midpoint temperature of unfolding decrease linearly with increasing guanidineHCl concentration, caused by the weak binding of denaturant. At room temperature, binding of 50-60 molecules guanidineHCl leads to a complete Apo A-1 unfolding. The entropy of unfolding decreases to a lesser extent than the unfolding enthalpy. Apo A-1 chemical unfolding is a dynamic multi-state equilibrium that is analysed with the Zimm-Bragg theory modified for chemical unfolding. The chemical Zimm-Bragg theory predicts the denaturant binding constant K D and the protein cooperativity σ. Chemical unfolding of Apo A-1 is two orders of magnitude less cooperative than thermal unfolding. The free energy of thermal unfolding is ~0.2 kcal/mol per amino acid residue and ~1.0 kcal/mol for chemical unfolding at room temperature. The Zimm-Bragg theory calculates conformational probabilities and the chemical Zimm-Bragg theory predicts stretches of α-helical segments in dynamic equilibrium, unfolding and refolding independently and fast. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Queensland Seasons

Atmospheric Science Data Center

2016-05-27

... are in turn influenced by vegetation structure, terrain and soil type, and by the different solar illumination conditions on the two dates. ... wavelenths is strongly scattered between the leaf layers of the dense canopies, and the influence of shadows between the tree ...

Soybean canopy reflectance as influenced by cultural practices. [West Lafayette, Indiana

NASA Technical Reports Server (NTRS)

Bauer, M. E. (Principal Investigator); Kollenkark, J. C.; Daughtry, C. S. T.

1981-01-01

Experiments were conducted at West Lafayette, Indiana in 1978 and 1979 to study the reflectance factor of soybean canopies as affected by differences in row width, population, planting date, cultivar and soil type. Reflectance factor data were acquired throughout the growing season with a LANDSAT-band radiometer. Agronomic data included plant height, leaf area index, development stage, total fresh and dry biomass, percent soil cover, and grain yield. The results indicate that row width, planting date, and cultivar influence the percent soil cover, leaf area index, and biomass present, which are in turn related to the multispectral reflectance. Additionally, the reflectance data were quite sensitive to the onset of senescence. Soil color and moisture were found to be important factors influencing the reflectance in single LANDSAT bands, but the near infrared/red reflectance ratio and the greeness transformation were less sensitive than the single bands to the soil background present.

Estimates of leaf area index from spectral reflectance of wheat under different cultural practices and solar angle

NASA Technical Reports Server (NTRS)

Asrar, G.; Kanemasu, E. T.; Yoshida, M.

1985-01-01

The influence of management practices and solar illumination angle on the leaf area index (LAI) was estimated from measurements of wheat canopy reflectance evaluated by two methods, a regression formula and an indirect technique. The date of planting and the time of irrigation in relation to the stage of plant growth were found to have significant effects on the development of leaves in spring wheat. A reduction in soil moisture adversely affected both the duration and magnitude of the maximum LAI for late planting dates. In general, water stress during vegetative stages resulted in a reduction in maximum LAI, while water stress during the reproductive period shortened the duration of green LAI in spring wheat. Canopy geometry and solar angle also affected the spectral properties of the canopies, and hence the estimated LAI. Increase in solar zenith angles resulted in a general increase in estimated LAI obtained from both methods.

Effects of nitrogen nutrition on the growth, yield and reflectance characteristics of corn canopies. [Purdue Agronomy Farm, Indiana

NASA Technical Reports Server (NTRS)

Bauer, M. E. (Principal Investigator); Walburg, G.; Daughtry, C. S. T.

1981-01-01

Spectral and agronomic measurements were collected from corn (Zea mays L.) canopies under four nitrogen treatment levels (0, 67, 134, and 202 kg/ha) on 11 dates during 1978 and 12 dates during 1979. Data were analyzed to determine the relationship between the spectral responses of canopies and their argonomic characteristics as well as the spectral separability of the four treatments. Red reflectance was increased, while the near infrared reflectance was decreased for canopies under nitrogen deprivation. Spectral differences between treatments were seen throughout each growing season. The near infrared/red reflectance ratio increased spectral treatment differences over those shown by single band reflectance measures. Of the spectral variables examined, the near infrared/red reflectance ratio most effectively separated the treatments. Differences in spectral response between treatments were attributed to varying soil cover, leaf area index, and leaf pigmentation values, all of which changed with N treatment.

Unfolding and unfoldability of digital pulses in the z-domain

NASA Astrophysics Data System (ADS)

Regadío, Alberto; Sánchez-Prieto, Sebastián

2018-04-01

The unfolding (or deconvolution) technique is used in the development of digital pulse processing systems applied to particle detection. This technique is applied to digital signals obtained by digitization of analog signals that represent the combined response of the particle detectors and the associated signal conditioning electronics. This work describes a technique to determine if the signal is unfoldable. For unfoldable signals the characteristics of the unfolding system (unfolder) are presented. Finally, examples of the method applied to real experimental setup are discussed.

An overlapping region between the two terminal folding units of the outer surface protein A (OspA) controls its folding behavior.

PubMed

Makabe, Koki; Nakamura, Takashi; Dhar, Debanjan; Ikura, Teikichi; Koide, Shohei; Kuwajima, Kunihiro

2018-04-27

Although many naturally occurring proteins consist of multiple domains, most studies on protein folding to date deal with single-domain proteins or isolated domains of multi-domain proteins. Studies of multi-domain protein folding are required for further advancing our understanding of protein folding mechanisms. Borrelia outer surface protein A (OspA) is a β-rich two-domain protein, in which two globular domains are connected by a rigid and stable single-layer β-sheet. Thus, OspA is particularly suited as a model system for studying the interplays of domains in protein folding. Here, we studied the equilibria and kinetics of the urea-induced folding-unfolding reactions of OspA probed with tryptophan fluorescence and ultraviolet circular dichroism. Global analysis of the experimental data revealed compelling lines of evidence for accumulation of an on-pathway intermediate during kinetic refolding and for the identity between the kinetic intermediate and a previously described equilibrium unfolding intermediate. The results suggest that the intermediate has the fully native structure in the N-terminal domain and the single layer β-sheet, with the C-terminal domain still unfolded. The observation of the productive on-pathway folding intermediate clearly indicates substantial interactions between the two domains mediated by the single-layer β-sheet. We propose that a rigid and stable intervening region between two domains creates an overlap between two folding units and can energetically couple their folding reactions. Copyright © 2018. Published by Elsevier Ltd.

Seasonality and phenology alter functional leaf traits.

PubMed

McKown, Athena D; Guy, Robert D; Azam, M Shofiul; Drewes, Eric C; Quamme, Linda K

2013-07-01

In plant ecophysiology, functional leaf traits are generally not assessed in relation to phenological phase of the canopy. Leaf traits measured in deciduous perennial species are known to vary between spring and summer seasons, but there is a knowledge gap relating to the late-summer phase marked by growth cessation and bud set occurring well before fall leaf senescence. The effects of phenology on canopy physiology were tested using a common garden of over 2,000 black cottonwood (Populus trichocarpa) individuals originating from a wide geographical range (44-60ºN). Annual phenological events and 12 leaf-based functional trait measurements were collected spanning the entire summer season prior to, and following, bud set. Patterns of seasonal trait change emerged by synchronizing trees using their date of bud set. In particular, photosynthetic, mass, and N-based traits increased substantially following bud set. Most traits were significantly different between pre-bud set and post-bud set phase trees, with many traits showing at least 25% alteration in mean value. Post-bud set, both the significance and direction of trait-trait relationships could be modified, with many relating directly to changes in leaf mass. In Populus, these dynamics in leaf traits throughout the summer season reflected a shift in whole plant physiology, but occurred long before the onset of leaf senescence. The marked shifts in measured trait values following bud set underscores the necessity to include phenology in trait-based ecological studies or large-scale phenotyping efforts, both at the local level and larger geographical scale.

Satellite-based phenology detection in broadleaf forests in South-Western Germany

NASA Astrophysics Data System (ADS)

Misra, Gourav; Buras, Allan; Menzel, Annette

2016-04-01

Many techniques exist for extracting phenological information from time series of satellite data. However, there have been only a few successful attempts to temporarily match satellite-derived observations with ground based phenological observations (Fisher et al., 2006; Hamunyela et al., 2013; Galiano et al., 2015). Such studies are primarily plagued with problems relating to shorter time series of satellite data including spatial and temporal resolution issues. A great challenge is to correlate spatially continuous and pixel-based satellite information with spatially discontinuous and point-based, mostly species-specific, ground observations of phenology. Moreover, the minute differences in phenology observed by ground volunteers might not be sufficient to produce changes in satellite-measured reflectance of vegetation, which also exposes the difference in the definitions of phenology (Badeck et al., 2004; White et al., 2014). In this study Start of Season (SOS) was determined for broadleaf forests at a site in south-western Germany using MODIS-sensor time series of Normalised Difference Vegetation Index (NDVI) data for the years covering 2001 to 2013. The NDVI time series raster data was masked for broadleaf forests using Corine Land Cover dataset, filtered and corrected for snow and cloud contaminations, smoothed with a Gaussian filter and interpolated to daily values. Several SOS techniques cited in literature, namely thresholds of amplitudes (20%, 50%, 60% and 75%), rates of change (1st, 2nd and 3rd derivative) and delayed moving average (DMA) were tested for determination of satellite SOS. The different satellite SOS were then compared with a species-rich ground based phenology information (e.g. understory leaf unfolding, broad leaf unfolding and greening of evergreen tree species). Working with all the pixels at a finer resolution, it is seen that the temporal trends in understory and broad leaf species are well captured. Initial analyses show promising results and suggest that different satellite SOS extraction techniques work well for specific phases of ground phenology information. More than half of the broadleaf pixels show an earliness in SOS which matches with the trend in ground phenology. References 1. F.-W. Badeck, A. Bondeau, K. Bottcher, D. Doktor, W. Lucht, J. Schaber, and S. Sitch, 2004, "Responses of spring phenology to climate change," New Phytologist, vol. 162, no. 2, pp. 295-309. 2. E. Hamunyela, J. Verbesselt, G. Roerink, and M. Herold, 2013, "Trends in Spring Phenology of Western European Deciduous Forests," Remote Sensing, vol. 5, no. 12, pp. 6159-6179. 3. V. F. Rodriguez-Galiano, J. Dash, and P. M. Atkinson, 2015, "Intercomparison of satellite sensor land surface phenology and ground phenology in Europe: Inter-annual comparison and modelling," Geophysical Research Letters, vol. 42, no. 7, pp. 2253-2260. 4. J. Fisher, J. Mustard, and M. Vadeboncoeur, 2006, "Green leaf phenology at Landsat resolution: Scaling from the field to the satellite," Remote Sensing of Environment, vol. 100, no. 2, pp. 265-279. 5. K. White, J. Pontius, and P. Schaberg, 2014, "Remote sensing of spring phenology in northeastern forests: A comparison of methods, field metrics and sources of uncertainty," Remote Sensing of Environment, vol. 148, pp. 97-107.

Model-data assimilation of multiple phenological observations to constrain and predict leaf area index.

PubMed

Viskari, Toni; Hardiman, Brady; Desai, Ankur R; Dietze, Michael C

2015-03-01

Our limited ability to accurately simulate leaf phenology is a leading source of uncertainty in models of ecosystem carbon cycling. We evaluate if continuously updating canopy state variables with observations is beneficial for predicting phenological events. We employed ensemble adjustment Kalman filter (EAKF) to update predictions of leaf area index (LAI) and leaf extension using tower-based photosynthetically active radiation (PAR) and moderate resolution imaging spectrometer (MODIS) data for 2002-2005 at Willow Creek, Wisconsin, USA, a mature, even-aged, northern hardwood, deciduous forest. The ecosystem demography model version 2 (ED2) was used as the prediction model, forced by offline climate data. EAKF successfully incorporated information from both the observations and model predictions weighted by their respective uncertainties. The resulting. estimate reproduced the observed leaf phenological cycle in the spring and the fall better than a parametric model prediction. These results indicate that during spring the observations contribute most in determining the correct bud-burst date, after which the model performs well, but accurately modeling fall leaf senesce requires continuous model updating from observations. While the predicted net ecosystem exchange (NEE) of CO2 precedes tower observations and unassimilated model predictions in the spring, overall the prediction follows observed NEE better than the model alone. Our results show state data assimilation successfully simulates the evolution of plant leaf phenology and improves model predictions of forest NEE.

Collision-Induced Dissociation of Electrosprayed Protein Complexes: An All-Atom Molecular Dynamics Model with Mobile Protons.

PubMed

Popa, Vlad; Trecroce, Danielle A; McAllister, Robert G; Konermann, Lars

2016-06-16

Electrospray ionization mass spectrometry (ESI-MS) has become an indispensable technique for examining noncovalent protein complexes. Collision-induced dissociation (CID) of these multiply protonated gaseous ions usually culminates in ejection of a single subunit with a disproportionately large amount of charge. Experiments suggest that this process involves subunit unfolding prior to separation from the residual complex, as well as H(+) migration onto the unravelling chain. Molecular dynamics (MD) simulations are a promising avenue for gaining detailed insights into these CID events. Unfortunately, typical MD algorithms do not allow for mobile protons. Here we address this limitation by implementing a strategy that combines atomistic force fields (such as OPLS/AA and CHARMM36) with a proton hopping algorithm, focusing on the tetrameric complexes transthyretin and streptavidin. Protons are redistributed over all acidic and basic sites in 20 ps intervals, subject to an energy function that reflects electrostatic interactions and proton affinities. Our simulations predict that nativelike conformers at the onset of collisional heating contain multiple salt bridges. Collisional heating initially causes subtle structural changes that lead to a gradual decline of these zwitterionic patterns. Many of the MD runs show gradual unfolding of a single subunit in conjunction with H(+) migration, culminating in subunit separation from the complex. However, there are also instances where two or more chains start to unfold simultaneously, giving rise to charge competition. The scission point where the "winning" subunit separates from the complex can be attained for different degrees of unfolding, giving rise to product ions in various charge states. The simulated product ion distributions are in close agreement with experimental CID data. Proton enrichment in the departing subunit is driven by charge-charge repulsion, but the combination of salt bridge depletion, charge migration, and proton affinity causes surprising compensation effects among the various energy terms. It appears that this work provides the most detailed account to date of the mechanism whereby noncovalent protein complexes disassemble during CID.

Direct Observation of Markovian Behavior of the Mechanical Unfolding of Individual Proteins

PubMed Central

Cao, Yi; Kuske, Rachel; Li, Hongbin

2008-01-01

Single-molecule force-clamp spectroscopy is a valuable tool to analyze unfolding kinetics of proteins. Previous force-clamp spectroscopy experiments have demonstrated that the mechanical unfolding of ubiquitin deviates from the generally assumed Markovian behavior and involves the features of glassy dynamics. Here we use single molecule force-clamp spectroscopy to study the unfolding kinetics of a computationally designed fast-folding mutant of the small protein GB1, which shares a similar β-grasp fold as ubiquitin. By treating the mechanical unfolding of polyproteins as the superposition of multiple identical Poisson processes, we developed a simple stochastic analysis approach to analyze the dwell time distribution of individual unfolding events in polyprotein unfolding trajectories. Our results unambiguously demonstrate that the mechanical unfolding of NuG2 fulfills all criteria of a memoryless Markovian process. This result, in contrast with the complex mechanical unfolding behaviors observed for ubiquitin, serves as a direct experimental demonstration of the Markovian behavior for the mechanical unfolding of a protein and reveals the complexity of the unfolding dynamics among structurally similar proteins. Furthermore, we extended our method into a robust and efficient pseudo-dwell-time analysis method, which allows one to make full use of all the unfolding events obtained in force-clamp experiments without categorizing the unfolding events. This method enabled us to measure the key parameters characterizing the mechanical unfolding energy landscape of NuG2 with improved precision. We anticipate that the methods demonstrated here will find broad applications in single-molecule force-clamp spectroscopy studies for a wide range of proteins. PMID:18375518

Protein unfolding in detergents: effect of micelle structure, ionic strength, pH, and temperature.

PubMed Central

Otzen, Daniel E

2002-01-01

The 101-residue monomeric protein S6 unfolds in the anionic detergent sodium dodecyl sulfate (SDS) above the critical micelle concentration, with unfolding rates varying according to two different modes. Our group has proposed that spherical micelles lead to saturation kinetics in unfolding (mode 1), while cylindrical micelles prevalent at higher SDS concentrations induce a power-law dependent increase in the unfolding rate (mode 2). Here I investigate in more detail how micellar properties affect protein unfolding. High NaCl concentrations, which induce cylindrical micelles, favor mode 2. This is consistent with our model, though other effects such as electrostatic screening cannot be discounted. Furthermore, unfolding does not occur in mode 2 in the cationic detergent LTAB, which is unable to form cylindrical micelles. A strong retardation of unfolding occurs at higher LTAB concentrations, possibly due to the formation of dead-end protein-detergent complexes. A similar, albeit much weaker, effect is seen in SDS in the absence of salt. Chymotrypsin inhibitor 2 exhibits the same modes of unfolding in SDS as S6, indicating that this type of protein unfolding is not specific for S6. The unfolding process in mode 1 has an activation barrier similar in magnitude to that in water, while the activation barrier in mode 2 is strongly concentration-dependent. The strong pH-dependence of unfolding in SDS and LTAB suggests that the rate of unfolding in anionic detergent is modulated by repulsion between detergent headgroups and anionic side chains, while cationic side chains modulate unfolding rates in cationic detergents. PMID:12324439

Electrostatic effects in unfolded staphylococcal nuclease

PubMed Central

Fitzkee, Nicholas C.; García-Moreno E, Bertrand

2008-01-01

Structure-based calculations of pK a values and electrostatic free energies of proteins assume that electrostatic effects in the unfolded state are negligible. In light of experimental evidence showing that this assumption is invalid for many proteins, and with increasing awareness that the unfolded state is more structured and compact than previously thought, a detailed examination of electrostatic effects in unfolded proteins is warranted. Here we address this issue with structure-based calculations of electrostatic interactions in unfolded staphylococcal nuclease. The approach involves the generation of ensembles of structures representing the unfolded state, and calculation of Coulomb energies to Boltzmann weight the unfolded state ensembles. Four different structural models of the unfolded state were tested. Experimental proton binding data measured with a variant of nuclease that is unfolded under native conditions were used to establish the validity of the calculations. These calculations suggest that weak Coulomb interactions are an unavoidable property of unfolded proteins. At neutral pH, the interactions are too weak to organize the unfolded state; however, at extreme pH values, where the protein has a significant net charge, the combined action of a large number of weak repulsive interactions can lead to the expansion of the unfolded state. The calculated pK a values of ionizable groups in the unfolded state are similar but not identical to the values in small peptides in water. These studies suggest that the accuracy of structure-based calculations of electrostatic contributions to stability cannot be improved unless electrostatic effects in the unfolded state are calculated explicitly. PMID:18227429

Effects of climate change on phenological trends and seed cotton yields in oasis of arid regions.

PubMed

Huang, Jian; Ji, Feng

2015-07-01

Understanding the effects of climatic change on phenological phases of cotton (Gossypium hirsutum L.) in oasis of arid regions may help optimize management schemes to increase productivity. This study assessed the impacts of climatic changes on the phenological phases and productivity of spring cotton. The results showed that climatic warming led the dates of sowing seed, seeding emergence, three-leaf, five-leaf, budding, anthesis, full bloom, cleft boll, boll-opening, boll-opening filling, and stop-growing become earlier by 24.42, 26.19, 24.75, 23.28, 22.62, 15.75, 14.58, 5.37, 2.85, 8.04, and 2.16 days during the period of 1981-2010, respectively. The growth period lengths from sowing seed to seeding emergence and from boll-opening to boll-opening filling were shortened by 1.76 and 5.19 days, respectively. The other growth period lengths were prolonged by 2-9.71 days. The whole growth period length was prolonged by 22.26 days. The stop-growing date was delayed by 2.49-3.46 days for every 1 °C rise in minimum, maximum, and mean temperatures; however, other development dates emerged earlier by 2.17-4.76 days. Rising temperatures during the stage from seeding emergence to three-leaf reduced seed cotton yields. However, rising temperatures increased seed cotton yields in the two stages from anthesis to cleft boll and from boll-opening filling to the stop-growing. Increasing accumulated temperatures (AT) had different impacts on different development stages. During the vegetative phase, rising AT led to reduced seed cotton yields, but rising AT during reproductive stage increased seed cotton yields. In conclusion, climatic warming helpfully obtained more seed cotton yields in oasis of arid regions in northwest China. Changing the sowing date is another way to enhance yields for climate change in the future.

Volatile components of vine leaves from two Portuguese grape varieties (Vitis vinifera L.), Touriga Nacional and Tinta Roriz, analysed by solid-phase microextraction.

PubMed

Fernandes, Bruno; Correia, Ana C; Cosme, Fernanda; Nunes, Fernando M; Jordão, António M

2015-01-01

The purpose of this work was to study the volatile composition of vine leaves and vine leaf infusion prepared from vine leaves collected at 30 and 60 days after grape harvest of two Vitis vinifera L. species. Eighteen volatile compounds were identified by gas chromatography-mass spectrometry in vine leaves and in vine leaf infusions. It was observed that the volatile compounds present in vine leaves are dependent on the time of harvest, with benzaldehyde being the major volatile present in vine leaves collected at 30 days after harvesting. There are significant differences in the volatile composition of the leaves from the two grape cultivars, especially in the sample collected at 60 days after grape harvest. This is not reflected in the volatile composition of the vine leaf infusion made from this two cultivars, the more important being the harvesting date for the volatile profile of vine leaf infusion than the vine leaves grape cultivar.

Impact of sampling parameters on the radical scavenging potential of olive (Olea europaea L.) leaves.

PubMed

Papoti, Vassiliki T; Tsimidou, Maria Z

2009-05-13

The impact of sampling parameters, that is, cultivar, leaf age, and sampling date, on the radical scavenging potential of olive leaf extracts was examined via the DPPH(*) and other assays. Total phenol content was estimated colorimetrically and by fluorometry, whereas phenol composition was assessed by RP-HPLC coupled with diode array, fluorometric, and MS detection systems. Oleuropein was not always the major leaf constituent. Considerable differences noted in individual phenol levels (hydroxytyrosol, oleuropein and other secoiridoids, verbascoside, and flavonoids) among samples were not reflected either in the total phenol content or in the radical scavenging potential of the extracts. It can be suggested that olive leaf is a robust source of radical scavengers throughout the year and that differentiation in the levels of individual components depends rather on sampling period than on cultivar or age. The latter does not present predictable regularity. Exploitation of all types of leaves expected in an olive tree shoot for the extraction of bioactive compounds is feasible.

Dynamic Engagement of Cognitive Control Modulates Recovery From Misinterpretation During Real-Time Language Processing.

PubMed

Hsu, Nina S; Novick, Jared M

2016-04-01

Speech unfolds swiftly, yet listeners keep pace by rapidly assigning meaning to what they hear. Sometimes, though, initial interpretations turn out to be wrong. How do listeners revise misinterpretations of language input moment by moment to avoid comprehension errors? Cognitive control may play a role by detecting when processing has gone awry and then initiating behavioral adjustments accordingly. However, no research to date has investigated a cause-and-effect interplay between cognitive-control engagement and the overriding of erroneous interpretations in real time. Using a novel cross-task paradigm, we showed that Stroop-conflict detection, which mobilizes cognitive-control procedures, subsequently facilitates listeners' incremental processing of temporarily ambiguous spoken instructions that induce brief misinterpretation. When instructions followed incongruent Stroop items, compared with congruent Stroop items, listeners' eye movements to objects in a scene reflected more transient consideration of the false interpretation and earlier recovery of the correct one. Comprehension errors also decreased. Cognitive-control engagement therefore accelerates sentence-reinterpretation processes, even as linguistic input is still unfolding. © The Author(s) 2016.

Tuning the free-energy landscape of a WW domain by temperature, mutation, and truncation

PubMed Central

Nguyen, Houbi; Jäger, Marcus; Moretto, Alessandro; Gruebele, Martin; Kelly, Jeffery W.

2003-01-01

The equilibrium unfolding of the Formin binding protein 28 (FBP) WW domain, a stable three-stranded β-sheet protein, can be described as reversible apparent two-state folding. Kinetics studied by laser temperature jump reveal a third state at temperatures below the midpoint of unfolding. The FBP free-energy surface can be tuned between three-state and two-state kinetics by changing the temperature, by truncation of the C terminus, or by selected point mutations. FBP WW domain is the smallest three-state folder studied to date and the only one that can be freely tuned between three-state and apparent two-state folding by several methods (temperature, truncation, and mutation). Its small size (28–37 residues), the availability of a quantitative reaction coordinate (φT), the fast folding time scale (10s of μs), and the tunability of the folding routes by small temperature or sequence changes make this system the ideal prototype for studying more subtle features of the folding free-energy landscape by simulations or analytical theory. PMID:12651955

Tuning the free-energy landscape of a WW domain by temperature, mutation, and truncation.

PubMed

Nguyen, Houbi; Jager, Marcus; Moretto, Alessandro; Gruebele, Martin; Kelly, Jeffery W

2003-04-01

The equilibrium unfolding of the Formin binding protein 28 (FBP) WW domain, a stable three-stranded beta-sheet protein, can be described as reversible apparent two-state folding. Kinetics studied by laser temperature jump reveal a third state at temperatures below the midpoint of unfolding. The FBP free-energy surface can be tuned between three-state and two-state kinetics by changing the temperature, by truncation of the C terminus, or by selected point mutations. FBP WW domain is the smallest three-state folder studied to date and the only one that can be freely tuned between three-state and apparent two-state folding by several methods (temperature, truncation, and mutation). Its small size (28-37 residues), the availability of a quantitative reaction coordinate (phi(T)), the fast folding time scale (10s of micros), and the tunability of the folding routes by small temperature or sequence changes make this system the ideal prototype for studying more subtle features of the folding free-energy landscape by simulations or analytical theory.

Studying the unfolding process of protein G and protein L under physical property space

PubMed Central

Zhao, Liling; Wang, Jihua; Dou, Xianghua; Cao, Zanxia

2009-01-01

Background The studies on protein folding/unfolding indicate that the native state topology is an important determinant of protein folding mechanism. The folding/unfolding behaviors of proteins which have similar topologies have been studied under Cartesian space and the results indicate that some proteins share the similar folding/unfolding characters. Results We construct physical property space with twelve different physical properties. By studying the unfolding process of the protein G and protein L under the property space, we find that the two proteins have the similar unfolding pathways that can be divided into three types and the one which with the umbrella-shape represents the preferred pathway. Moreover, the unfolding simulation time of the two proteins is different and protein L unfolding faster than protein G. Additionally, the distributing area of unfolded state ensemble of protein L is larger than that of protein G. Conclusion Under the physical property space, the protein G and protein L have the similar folding/unfolding behaviors, which agree with the previous results obtained from the studies under Cartesian coordinate space. At the same time, some different unfolding properties can be detected easily, which can not be analyzed under Cartesian coordinate space. PMID:19208146

Exploring Early Stages of the Chemical Unfolding of Proteins at the Proteome Scale

PubMed Central

Candotti, Michela; Pérez, Alberto; Ferrer-Costa, Carles; Rueda, Manuel; Meyer, Tim; Gelpí, Josep Lluís; Orozco, Modesto

2013-01-01

After decades of using urea as denaturant, the kinetic role of this molecule in the unfolding process is still undefined: does urea actively induce protein unfolding or passively stabilize the unfolded state? By analyzing a set of 30 proteins (representative of all native folds) through extensive molecular dynamics simulations in denaturant (using a range of force-fields), we derived robust rules for urea unfolding that are valid at the proteome level. Irrespective of the protein fold, presence or absence of disulphide bridges, and secondary structure composition, urea concentrates in the first solvation shell of quasi-native proteins, but with a density lower than that of the fully unfolded state. The presence of urea does not alter the spontaneous vibration pattern of proteins. In fact, it reduces the magnitude of such vibrations, leading to a counterintuitive slow down of the atomic-motions that opposes unfolding. Urea stickiness and slow diffusion is, however, crucial for unfolding. Long residence urea molecules placed around the hydrophobic core are crucial to stabilize partially open structures generated by thermal fluctuations. Our simulations indicate that although urea does not favor the formation of partially open microstates, it is not a mere spectator of unfolding that simply displaces to the right of the folded←→unfolded equilibrium. On the contrary, urea actively favors unfolding: it selects and stabilizes partially unfolded microstates, slowly driving the protein conformational ensemble far from the native one and also from the conformations sampled during thermal unfolding. PMID:24348236

Distribution, transition and thermodynamic stability of protein conformations in the denaturant-induced unfolding of proteins.

PubMed

Bian, Liujiao; Ji, Xu

2014-01-01

Extensive and intensive studies on the unfolding of proteins require appropriate theoretical model and parameter to clearly illustrate the feature and characteristic of the unfolding system. Over the past several decades, four approaches have been proposed to describe the interaction between proteins and denaturants, but some ambiguity and deviations usually occur in the explanation of the experimental data. In this work, a theoretical model was presented to show the dependency of the residual activity ratio of the proteins on the molar denaturant concentration. Through the characteristic unfolding parameters ki and Δmi in this model, the distribution, transition and thermodynamic stability of protein conformations during the unfolding process can be quantitatively described. This model was tested with the two-state unfolding of bovine heart cytochrome c and the three-state unfolding of hen egg white lysozyme induced by both guanidine hydrochloride and urea, the four-state unfolding of bovine carbonic anhydrase b induced by guanidine hydrochloride and the unfolding of some other proteins induced by denaturants. The results illustrated that this model could be used accurately to reveal the distribution and transition of protein conformations in the presence of different concentrations of denaturants and to evaluate the unfolding tendency and thermodynamic stability of different conformations. In most denaturant-induced unfolding of proteins, the unfolding became increasingly hard in next transition step and the proteins became more unstable as they attained next successive stable conformation. This work presents a useful method for people to study the unfolding of proteins and may be used to describe the unfolding and refolding of other biopolymers induced by denaturants, inducers, etc.

Monitoring of cadmium in "on" and "off" date palms.

PubMed

Pillay, A E; Williams, J R; El Mardi, M O; Hassan, S M; Al-Hamdi, A

2002-09-01

The study demonstrated that the mature dates we investigated were considered safe for human consumption. However, our findings revealed that much of the early fruit and leaves, which appeared at the advent of the fruiting season, possessed elevated levels of cadmium (Cd) that could accumulate in the food chain and thus impact adversely on the environment. In addition, animal feed for livestock contains date leaves, which should be restricted to the mature ones. The monitoring of cadmium in date palms is of considerable interest to environmental science and the thrust of this work, therefore, involved measurement, by ICP, of Cd in dates and corresponding leaf specimens, and evaluation of its distribution during the developmental stages of the fruiting season. Thirty-six date samples and 36 leaf specimens of the Fard cultivar were collected from "on" and "off" date palms during the Kimri, Bisir and Rutab stages of the fruiting season and subjected to suitable digestion procedures. Sample masses of typically 1 g (dry weight) were prepared in 25 ml dilute acid solution and investigated for trace levels of Cd by ICP-AES. Special attention was paid to contamination and the validation of our methodology. The Cd "threshold" in our study was 50 ng/g, in keeping with the levels of tolerance appearing in the literature. For the dates we found elevated levels of Cd [> 50 ng/g] in most of the samples, for both categories of "on" and "off" trees, during the Kimri stage. In the case of the leaves, the "on" samples revealed significant values up to 125 ng/g in Bisir, in some cases, but the trend for the "off" trees remained the same with the highest levels [> 100 ng/g] recorded during Kimri. Safe levels were attained during Rutab for all specimens. The possibility of a connection between Cd toxicity and the alternate-bearing phenomenon is discussed and this could be the subject of future interest.

Leaf rust of cultivated barley: pathology and control.

PubMed

Park, Robert F; Golegaonkar, Prashant G; Derevnina, Lida; Sandhu, Karanjeet S; Karaoglu, Haydar; Elmansour, Huda M; Dracatos, Peter M; Singh, Davinder

2015-01-01

Leaf rust of barley is caused by the macrocyclic, heteroecious rust pathogen Puccinia hordei, with aecia reported from selected species of the genera Ornithogalum, Leopoldia, and Dipcadi, and uredinia and telia occurring on Hordeum vulgare, H. vulgare ssp. spontaneum, Hordeum bulbosum, and Hordeum murinum, on which distinct parasitic specialization occurs. Although Puccinia hordei is sporadic in its occurrence, it is probably the most common and widely distributed rust disease of barley. Leaf rust has increased in importance in recent decades in temperate barley-growing regions, presumably because of more intensive agricultural practices. Although total crop loss does not occur, under epidemic conditions yield reductions of up to 62% have been reported in susceptible varieties. Leaf rust is primarily controlled by the use of resistant cultivars, and, to date, 21 seedling resistance genes and two adult plant resistance (APR) genes have been identified. Virulence has been detected for most seedling resistance genes but is unknown for the APR genes Rph20 and Rph23. Other potentially new sources of APR have been reported, and additivity has been described for some of these resistances. Approaches to achieving durable resistance to leaf rust in barley are discussed.

Tuning Transpiration by Interfacial Solar Absorber-Leaf Engineering.

PubMed

Zhuang, Shendong; Zhou, Lin; Xu, Weichao; Xu, Ning; Hu, Xiaozhen; Li, Xiuqiang; Lv, Guangxin; Zheng, Qinghui; Zhu, Shining; Wang, Zhenlin; Zhu, Jia

2018-02-01

Plant transpiration, a process of water movement through a plant and its evaporation from aerial parts especially leaves, consumes a large component of the total continental precipitation (≈48%) and significantly influences global water distribution and climate. To date, various chemical and/or biological explorations have been made to tune the transpiration but with uncertain environmental risks. In recent years, interfacial solar steam/vapor generation is attracting a lot of attention for achieving high energy transfer efficiency. Various optical and thermal designs at the solar absorber-water interface for potential applications in water purification, seawater desalination, and power generation appear. In this work, the concept of interfacial solar vapor generation is extended to tunable plant transpiration by showing for the first time that the transpiration efficiency can also be enhanced or suppressed through engineering the solar absorber-leaf interface. By tuning the solar absorption of membrane in direct touch with green leaf, surface temperature of green leaf will change accordingly because of photothermal effect, thus the transpiration efficiency as well as temperature and relative humidity in the surrounding environment will be tuned. This tunable transpiration by interfacial absorber-leaf engineering can open an alternative avenue to regulate local atmospheric temperature, humidity, and eventually hydrologic cycle.

Gas exchange and leaf aging in an evergreen oak: causes and consequences for leaf carbon balance and canopy respiration.

PubMed

Rodríguez-Calcerrada, Jesús; Limousin, Jean-Marc; Martin-StPaul, Nicolas K; Jaeger, Carsten; Rambal, Serge

2012-04-01

Leaves of Mediterranean evergreens experience large variations in gas exchange rates over their life span due to aging and seasonally changing environmental conditions. Accounting for the changing respiratory physiology of leaves over time will help improve estimations of leaf and whole-plant carbon balances. Here we examined seasonal variations in light-saturated net CO(2) assimilation (A(max)), dark respiration (R(d)) and the proportional change in R(d) per 10 °C change in temperature (Q(10) of R(d)) in previous-year (PY) and current-year (CY) leaves of the broadleaved evergreen tree Quercus ilex L. A(max) and R(d) were lower in PY than in CY leaves. Differences in nitrogen between cohorts only partly explained such differences, and rates of A(max) and R(d) expressed per unit of leaf nitrogen were still significantly different between cohorts. The decline in A(max) in PY leaves did not result in the depletion of total non-structural carbohydrates, whose concentration was in fact higher in PY than CY leaves. Leaf-level carbon balance modeled from gas exchange data was positive at all ages. Q(10) of R(d) did not differ significantly between leaf cohorts; however, failure to account for distinct R(d) between cohorts misestimated canopy leaf respiration by 13% across dates when scaling up leaf measurements to the canopy. In conclusion, the decline in A(max) in old leaves that are close to or exceed their mean life span does not limit the availability of carbohydrates, which are probably needed to sustain new growth, as well as R(d) and nutrient resorption during senescence. Accounting for leaf age as a source of variation of R(d) improves the estimation of foliar respiratory carbon release at the stand scale.

Folivory and disease occurrence on Ludwigia hexapetala in Guntersville Reservoir

USDA-ARS?s Scientific Manuscript database

We report leaf feeding, disease occurrence and associated indigenous herbivore/fungal pathogen communities on the introduced wetland species Ludwigia hexapetala at Guntersville Reservoir, AL. Plant populations were sampled on three dates from May to September, 2014. A complex of indigenous herbivore...

Phytohormones and microRNAs as sensors and regulators of leaf senescence: assigning macro roles to small molecules.

PubMed

Sarwat, Maryam; Naqvi, Afsar Raza; Ahmad, Parvaiz; Ashraf, Muhammad; Akram, Nudrat Aisha

2013-12-01

Ageing or senescence is an intricate and highly synchronized developmental phase in the life of plant parts including leaf. Senescence not only means death of a plant part, but during this process, different macromolecules undergo degradation and the resulting components are transported to other parts of the plant. During the period from when a leaf is young and green to the stage when it senesces, a multitude of factors such as hormones, environmental factors and senescence associated genes (SAGs) are involved. Plant hormones including salicylic acid, abscisic acid, jasmonic acid and ethylene advance leaf senescence, whereas others like cytokinins, gibberellins, and auxins delay this process. The environmental factors which generally affect plant development and growth, can hasten senescence, the examples being nutrient dearth, water stress, pathogen attack, radiations, high temperature and light intensity, waterlogging, and air, water or soil contamination. Other important influences include carbohydrate accumulation and high carbon/nitrogen level. To date, although several genes involved in this complex process have been identified, still not much information exists in the literature on the signalling mechanism of leaf senescence. Now, the Arabidopsis mutants have paved our way and opened new vistas to elucidate the signalling mechanism of leaf senescence for which various mutants are being utilized. Recent studies demonstrating the role of microRNAs in leaf senescence have reinforced our knowledge of this intricate process. This review provides a comprehensive and critical analysis of the information gained particularly on the roles of several plant growth regulators and microRNAs in regulation of leaf senescence. Copyright © 2013 Elsevier Inc. All rights reserved.

Amyloidogenesis of Natively Unfolded Proteins

PubMed Central

Uversky, Vladimir N.

2009-01-01

Aggregation and subsequent development of protein deposition diseases originate from conformational changes in corresponding amyloidogenic proteins. The accumulated data support the model where protein fibrillogenesis proceeds via the formation of a relatively unfolded amyloidogenic conformation, which shares many structural properties with the pre-molten globule state, a partially folded intermediate first found during the equilibrium and kinetic (un)folding studies of several globular proteins and later described as one of the structural forms of natively unfolded proteins. The flexibility of this structural form is essential for the conformational rearrangements driving the formation of the core cross-beta structure of the amyloid fibril. Obviously, molecular mechanisms describing amyloidogenesis of ordered and natively unfolded proteins are different. For ordered protein to fibrillate, its unique and rigid structure has to be destabilized and partially unfolded. On the other hand, fibrillogenesis of a natively unfolded protein involves the formation of partially folded conformation; i.e., partial folding rather than unfolding. In this review recent findings are surveyed to illustrate some unique features of the natively unfolded proteins amyloidogenesis. PMID:18537543

The unfolding effects on the protein hydration shell and partial molar volume: a computational study.

PubMed

Del Galdo, Sara; Amadei, Andrea

2016-10-12

In this paper we apply the computational analysis recently proposed by our group to characterize the solvation properties of a native protein in aqueous solution, and to four model aqueous solutions of globular proteins in their unfolded states thus characterizing the protein unfolded state hydration shell and quantitatively evaluating the protein unfolded state partial molar volumes. Moreover, by using both the native and unfolded protein partial molar volumes, we obtain the corresponding variations (unfolding partial molar volumes) to be compared with the available experimental estimates. We also reconstruct the temperature and pressure dependence of the unfolding partial molar volume of Myoglobin dissecting the structural and hydration effects involved in the process.

Leaf angle, tree species, and the functioning of broadleaf deciduous forest ecosystems

NASA Astrophysics Data System (ADS)

McNeil, B. E.; Brzostek, E. R.; Fahey, R. T.; King, C. J.; Flamenco, E. A.; Rescorl, S.; Erazo, D.; Heimerl, T.

2016-12-01

The effects of temperate forests on the global cycles of carbon, water, and energy depends strongly on how individual tree species adjust to the novel environmental conditions of the Anthropocene. Here, we seek to identify and understand ecological variability in one important component of tree canopies, the inclination angles of leaves. Leaf angle has important effects on forest albedo, photosynthesis, and evapotranspiration, but there is relatively little data to constrain the many models that include (or perhaps should include) this essential aspect of canopy architecture. We employ a relatively new technique for using an electronic protractor to measure leaf angles from leveled digital photographs. From a suite of observation platforms (e.g. UAVs, eddy flux towers, old fire towers) in Connecticut, Indiana, Maryland, Michigan, Pennsylvania, and West Virginia, USA, we have measured leaf angles periodically throughout the 2014, 2015, and 2016 growing seasons. Based on over 25,000 measurements taken from 15 tree species, we find highly significant differences in mean leaf angle by canopy position, tree species, location, and observation date. In addition to replicating findings where upper-canopy sun leaves are more vertical than lower-canopy shade leaves, our analysis on sun leaves also finds other ecologically meaningful differences. For instance, we find that the mesic, shade tolerant sugar maple had significantly more horizontal leaf angles than drought-resistant species such as white oak. Species also appear to have unique patterns of leaf angle phenology, with most species tending toward more vertical leaf angles during droughty conditions later in the year. We discuss these empirical results in light of an emerging theoretical framework that positions leaf angle as a functional trait. Like leaf traits such as %N or SLA, we suggest that leaf angle is an essential part of the adaptive resource strategy of each tree species. Finally, by linking our leaf angle data to new observations of spatial and temporal variations in near infrared reflectance measured from UAV, airborne, and satellite sensors, we highlight how species-specific patterns of leaf angle phenology could provide a new mechanism to better constrain model predictions of energy, water, and carbon fluxes from temperate forests.

Lethal factor unfolding is the most force-dependent step of anthrax toxin translocation

PubMed Central

Thoren, Katie L.; Worden, Evan J.; Yassif, Jaime M.; Krantz, Bryan A.

2009-01-01

Cellular compartmentalization requires machinery capable of translocating polypeptides across membranes. In many cases, transported proteins must first be unfolded by means of the proton motive force and/or ATP hydrolysis. Anthrax toxin, which is composed of a channel-forming protein and two substrate proteins, is an attractive model system to study translocation-coupled unfolding, because the applied driving force can be externally controlled and translocation can be monitored directly by using electrophysiology. By controlling the driving force and introducing destabilizing point mutations in the substrate, we identified the barriers in the transport pathway, determined which barrier corresponds to protein unfolding, and mapped how the substrate protein unfolds during translocation. In contrast to previous studies, we find that the protein's structure next to the signal tag is not rate-limiting to unfolding. Instead, a more extensive part of the structure, the amino-terminal β-sheet subdomain, must disassemble to cross the unfolding barrier. We also find that unfolding is catalyzed by the channel's phenylalanine-clamp active site. We propose a broad molecular mechanism for translocation-coupled unfolding, which is applicable to both soluble and membrane-embedded unfolding machines. PMID:19926859

Solvent sensitivity of protein unfolding: dynamical study of chicken villin headpiece subdomain in water-ethanol binary mixture.

PubMed

Ghosh, Rikhia; Roy, Susmita; Bagchi, Biman

2013-12-12

We carry out a series of long atomistic molecular dynamics simulations to study the unfolding of a small protein, chicken villin headpiece (HP-36), in water-ethanol (EtOH) binary mixture. The prime objective of this work is to explore the sensitivity of protein unfolding dynamics toward increasing concentration of the cosolvent and unravel essential features of intermediates formed in search of a dynamical pathway toward unfolding. In water-ethanol binary mixtures, HP-36 is found to unfold partially, under ambient conditions, that otherwise requires temperature as high as ∼600 K to denature in pure aqueous solvent. However, an interesting course of pathway is observed to be followed in the process, guided by the formation of unique intermediates. The first step of unfolding is essentially the separation of the cluster formed by three hydrophobic (phenylalanine) residues, namely, Phe-7, Phe-11, and Phe-18, which constitute the hydrophobic core, thereby initiating melting of helix-2 of the protein. The initial steps are similar to temperature-induced unfolding as well as chemical unfolding using DMSO as cosolvent. Subsequent unfolding steps follow a unique path. As water-ethanol shows composition-dependent anomalies, so do the details of unfolding dynamics. With an increase in cosolvent concentration, different partially unfolded intermediates are found to be formed. This is reflected in a remarkable nonmonotonic composition dependence of several order parameters, including fraction of native contacts and protein-solvent interaction energy. The emergence of such partially unfolded states can be attributed to the preferential solvation of the hydrophobic residues by the ethyl groups of ethanol. We further quantify the local dynamics of unfolding by using a Marcus-type theory.

Data Unfolding with Wiener-SVD Method

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

Tang, W.; Li, X.; Qian, X.

Here, data unfolding is a common analysis technique used in HEP data analysis. Inspired by the deconvolution technique in the digital signal processing, a new unfolding technique based on the SVD technique and the well-known Wiener filter is introduced. The Wiener-SVD unfolding approach achieves the unfolding by maximizing the signal to noise ratios in the effective frequency domain given expectations of signal and noise and is free from regularization parameter. Through a couple examples, the pros and cons of the Wiener-SVD approach as well as the nature of the unfolded results are discussed.

Data Unfolding with Wiener-SVD Method

DOE PAGES

Tang, W.; Li, X.; Qian, X.; ...

2017-10-04

Here, data unfolding is a common analysis technique used in HEP data analysis. Inspired by the deconvolution technique in the digital signal processing, a new unfolding technique based on the SVD technique and the well-known Wiener filter is introduced. The Wiener-SVD unfolding approach achieves the unfolding by maximizing the signal to noise ratios in the effective frequency domain given expectations of signal and noise and is free from regularization parameter. Through a couple examples, the pros and cons of the Wiener-SVD approach as well as the nature of the unfolded results are discussed.

Variability common to first leaf dates and snowpack in the western conterminous United States

USGS Publications Warehouse

McCabe, Gregory J.; Betancourt, Julio L.; Pederson, Gregory T.; Schwartz, Mark D.

2013-01-01

Singular value decomposition is used to identify the common variability in first leaf dates (FLDs) and 1 April snow water equivalent (SWE) for the western United States during the period 1900–2012. Results indicate two modes of joint variability that explain 57% of the variability in FLD and 69% of the variability in SWE. The first mode of joint variability is related to widespread late winter–spring warming or cooling across the entire west. The second mode can be described as a north–south dipole in temperature for FLD, as well as in cool season temperature and precipitation for SWE, that is closely correlated to the El Niño–Southern Oscillation. Additionally, both modes of variability indicate a relation with the Pacific–North American atmospheric pattern. These results indicate that there is a substantial amount of common variance in FLD and SWE that is related to large-scale modes of climate variability.

Nanomechanics of Protein Unfolding outside Protease Nanopores

NASA Astrophysics Data System (ADS)

Luan, Binquan; Zhou, Ruhong

Protein folding and unfolding have been the subject of active research for decades. Most of previous studies in protein unfolding were focused on temperature, chemical and/or force (such as in AFM) induced denaturations. Recent studies on the functional roles of proteasomes (such as ClpXP) revealed a novel unfolding process in cell, during which a target protein is mechanically unfolded and pulled into a confined, pore-like geometry for degradation. While the proteasome nanomachine has been extensively studied, the mechanism for unfolding proteins with the proteasome pore is still poorly understood. Here, we investigate the mechanical unfolding process of ubiquitin with (or really outside) an idealized proteasome pore, and compare such process with that in the AFM pulling experiment. Unexpectedly, the required force by a proteosome can be much smaller than that by the AFM. Simulation results also unveiled different nanomechanics, tearing fracture vs. shearing friction, in these two distinct types of mechanical unfoldings.

Linking belowground and aboveground phenology in two boreal forests in Northeast China.

PubMed

Du, Enzai; Fang, Jingyun

2014-11-01

The functional equilibrium between roots and shoots suggests an intrinsic linkage between belowground and aboveground phenology. However, much less understanding of belowground phenology hinders integrating belowground and aboveground phenology. We measured root respiration (Ra) as a surrogate for root phenology and integrated it with observed leaf phenology and radial growth in a birch (Betula platyphylla)-aspen (Populus davidiana) forest and an adjacent larch (Larix gmelinii) forest in Northeast China. A log-normal model successfully described the seasonal variations of Ra and indicated the initiation, termination and peak date of root phenology. Both root phenology and leaf phenology were highly specific, with a later onset, earlier termination, and shorter period of growing season for the pioneer tree species (birch and aspen) than the dominant tree species (larch). Root phenology showed later initiation, later peak and later termination dates than leaf phenology. An asynchronous correlation of Ra and radial growth was identified with a time lag of approximately 1 month, indicating aprioritization of shoot growth. Furthermore, we found that Ra was strongly correlated with soil temperature and air temperature, while radial growth was only significantly correlated with air temperature, implying a down-regulating effect of temperature. Our results indicate different phenologies between pioneer and dominant species and support a down-regulation hypothesis of plant phenology which can be helpful in understanding forest dynamics in the context of climate change.

Dating violence, bullying, and sexual harassment: longitudinal profiles and transitions over time.

PubMed

Miller, Shari; Williams, Jason; Cutbush, Stacey; Gibbs, Deborah; Clinton-Sherrod, Monique; Jones, Sarah

2013-04-01

Although there is growing recognition of the problem of dating violence, little is known about how it unfolds among young adolescents who are just beginning to date. This study examined classes (subgroups) and transitions between classes over three time points based on dating violence, bullying, and sexual harassment perpetration and victimization experienced by youth. The sample was ethnically diverse, consisting of 795 seventh-grade students from schools that were part of a multi-site, longitudinal evaluation of a dating violence initiative (50 % female; 27 % White, 32 % African American, 25 % Latino, 16 % other or multiple races). Results from latent transition analyses revealed five classes of students with distinct behavioral profiles: multi-problem (victimization and perpetration), bullying and sexual harassment (victimization and perpetration), bullying (victimization and perpetration) and sexual harassment (victimization only), bullying (victimization and perpetration), and a least problem group. The majority of classes were characterized by reports of both perpetration and victimization for at least one behavior. Girls were more likely to be in the less problematic classes. Class membership was fairly stable across the three time points. When students transitioned to a different class, the shift was most often from a more problematic to a less problematic class, particularly for girls. The findings support understanding dating violence within a dynamic, developmental process that recognizes related behaviors within and across individuals. Overall, the findings highlight the utility of person-oriented approaches to enhance our understanding of longitudinal profiles and transitions over time for dating violence and related behaviors.

Estimating cotton nitrogen nutrition status using leaf greenness and ground cover information

USDA-ARS?s Scientific Manuscript database

Assessing nitrogen (N) status is important from economic and environmental standpoints. To date, many spectral indices to estimate cotton chlorophyll or N content have been purely developed using statistical analysis approach where they are often subject to site-specific problems. This study describ...

Structure-function insights of membrane and soluble proteins revealed by electron crystallography.

PubMed

Dreaden, Tina M; Devarajan, Bharanidharan; Barry, Bridgette A; Schmidt-Krey, Ingeborg

2013-01-01

Electron crystallography is emerging as an important method in solving protein structures. While it has found extensive applications in the understanding of membrane protein structure and function at a wide range of resolutions, from revealing oligomeric arrangements to atomic models, electron crystallography has also provided invaluable information on the soluble α/β-tubulin which could not be obtained by any other method to date. Examples of critical insights from selected structures of membrane proteins as well as α/β-tubulin are described here, demonstrating the vast potential of electron crystallography that is first beginning to unfold.

Remote sensing and the optical properties of the narrow cylindrical leaves of Juncus roemerianus

USGS Publications Warehouse

Ramsey, Elijah W.; Rangoonwala, A.

2004-01-01

To develop a more complete foundation for remote sensing of the marsh grass Juncus roemerianus, we measured the optical properties of its cylindrical leaves at sites of different canopy height, biomass composition and amount, and connectivity to ocean flushing. To measure the leaf optical properties, we adapted a technique used for conifer needles. After establishing the reliability and limits of the adapted technique to the wider J.roemerianus leaves, mean transmittance and reflectance spectra were compared to associated leaf diameters from two dates in 1999 and 2002 and at each site. Transmittance was inversely related to leaf diameter. Mean transmittance and reflectance generated from reoccupation of many field sites in 2002 indicated little or no difference in transmittance between years, a slight reflectance difference in the visible (<2%) and a slightly higher reflectance difference in the near infrared (NIR) (<4%). Site comparison indicated limited ability to separate leaf transmittance but not reflectance by marsh type (e.g., low, medium, high) or biomass. Excluding one outlier, we found leaf transmittances could be adequately represented as 1% ?? 0.2% in the visible and 9% ?? 1% in the NIR and leaf reflectances represented from 14% to 16% in the visible and 71% to 75% in the NIR (the reflectance ranges represent 1999 and 2002 means). Reflectance and transmittance spectra associated with the dead J. roemerianus leaves displayed a spectrally flat increase from the visible to the NIR wavelengths. In total, we documented the atypical optical properties of the cylindrical J. roemerianus leaves and showed that to a first approximation, single means could represent leaf transmittance and visible leaf reflectance across all marsh zones and, after accounting for sample standardization, possibly the NIR reflectance as well.

Folding in and out: passive morphing in flapping wings.

PubMed

Stowers, Amanda K; Lentink, David

2015-03-25

We present a new mechanism for passive wing morphing of flapping wings inspired by bat and bird wing morphology. The mechanism consists of an unactuated hand wing connected to the arm wing with a wrist joint. Flapping motion generates centrifugal accelerations in the hand wing, forcing it to unfold passively. Using a robotic model in hover, we made kinematic measurements of unfolding kinematics as functions of the non-dimensional wingspan fold ratio (2-2.5) and flapping frequency (5-17 Hz) using stereo high-speed cameras. We find that the wings unfold passively within one to two flaps and remain unfolded with only small amplitude oscillations. To better understand the passive dynamics, we constructed a computer model of the unfolding process based on rigid body dynamics, contact models, and aerodynamic correlations. This model predicts the measured passive unfolding within about one flap and shows that unfolding is driven by centrifugal acceleration induced by flapping. The simulations also predict that relative unfolding time only weakly depends on flapping frequency and can be reduced to less than half a wingbeat by increasing flapping amplitude. Subsequent dimensional analysis shows that the time required to unfold passively is of the same order of magnitude as the flapping period. This suggests that centrifugal acceleration can drive passive unfolding within approximately one wingbeat in small and large wings. Finally, we show experimentally that passive unfolding wings can withstand impact with a branch, by first folding and then unfolding passively. This mechanism enables flapping robots to squeeze through clutter without sophisticated control. Passive unfolding also provides a new avenue in morphing wing design that makes future flapping morphing wings possibly more energy efficient and light-weight. Simultaneously these results point to possible inertia driven, and therefore metabolically efficient, control strategies in bats and birds to morph or recover within a beat.

Intercepted photosynthetically active radiation in wheat canopies estimated by spectral reflectance. [Phoenix, Arizona

NASA Technical Reports Server (NTRS)

Hatfield, J. L.; Asrar, G.; Kanemasu, E. T.

1982-01-01

The interception of photosynthetically active radiation (PAR) was evaluated relative to greenness and normalized difference (MSS 7-5/7+5) for five planting dates of wheat for 1978-79 and 1979-80 in Phoenix. Intercepted PAR was calculated from a model driven by leaf area index and stage of growth. Linear relationships were found between greenness and normalized difference with a separate model representing growth and senescence of the crop. Normalized difference was a significantly better model and would be easier to apply than the empirically derived greenness parameter. For the leaf area growth portion of the season the model between PAR interception and normalized difference was the same over years, however, for the leaf senescence the models showed more variability due to the lack of data on measured interception in sparse canopies. Normalized difference could be used to estimate PAR interception directly for crop growth models.

Using circular dichroism collected as a function of temperature to determine the thermodynamics of protein unfolding and binding interactions

PubMed Central

Greenfield, Norma J.

2009-01-01

Circular dichroism (CD) is an excellent spectroscopic technique for following the unfolding and folding of proteins as a function of temperature. One of its principal applications is to determine the effects of mutations and ligands on protein and polypeptide stability If the change in CD as a function of temperature is reversible, analysis of the data may be used to determined the van't Hoff enthalpy (ΔH) and entropy (ΔS) of unfolding, the midpoint of the unfolding transition (TM) and the free energy (ΔG) of unfolding. Binding constants of protein-protein and protein-ligand interactions may also be estimated from the unfolding curves. Analysis of CD spectra obtained as a function of temperature is also useful to determine whether a protein has unfolding intermediates. Measurement of the spectra of five folded proteins and their unfolding curves at a single wavelength takes approximately eight hours. PMID:17406506

Protein unfolding under isometric tension-what force can integrins generate, and can it unfold FNIII domains?

PubMed

Erickson, Harold P

2017-02-01

Extracellular matrix fibrils of fibronectin (FN) are highly elastic, and are typically stretched three to four times their relaxed length. The mechanism of stretching has been controversial, in particular whether it involves tension-induced unfolding of FNIII domains. Recent studies have found that ∼5pN is the threshold isometric force for unfolding various protein domains. FNIII domains should therefore not be unfolded until the tension approaches 5pN. Integrins have been reported to generate forces ranging from 1 to >50pN, but I argue that studies reporting 1-2pN are the most convincing. This is not enough to unfold FNIII domains. Even if domains were unfolded, 2pN would only extend the worm-like-chain to about twice the length of the folded domain. Overall I conclude that stretching FN matrix fibrils involves primarily the compact to extended conformational change of FN dimers, with minimal contribution from unfolding FNIII domains. Copyright © 2016 Elsevier Ltd. All rights reserved.

Point Mutations in Membrane Proteins Reshape Energy Landscape and Populate Different Unfolding Pathways

PubMed Central

Sapra, K. Tanuj; Balasubramanian, G. Prakash; Labudde, Dirk; Bowie, James U.; Muller, Daniel J.

2009-01-01

Using single-molecule force spectroscopy, we investigated the effect of single point mutations on the energy landscape and unfolding pathways of the transmembrane protein bacteriorhodopsin. We show that the unfolding energy barriers in the energy landscape of the membrane protein followed a simple two-state behavior and represent a manifestation of many converging unfolding pathways. Although the unfolding pathways of wild-type and mutant bacteriorhodopsin did not change, indicating the presence of same ensemble of structural unfolding intermediates, the free energies of the rate-limiting transition states of the bacteriorhodopsin mutants decreased as the distance of those transition states to the folded intermediate states decreased. Thus, all mutants exhibited Hammond behavior and a change in the free energies of the intermediates along the unfolding reaction coordinate and, consequently, their relative occupancies. This is the first experimental proof showing that point mutations can reshape the free energy landscape of a membrane protein and force single proteins to populate certain unfolding pathways over others. PMID:18191146

Prediction of change in protein unfolding rates upon point mutations in two state proteins.

PubMed

Chaudhary, Priyashree; Naganathan, Athi N; Gromiha, M Michael

2016-09-01

Studies on protein unfolding rates are limited and challenging due to the complexity of unfolding mechanism and the larger dynamic range of the experimental data. Though attempts have been made to predict unfolding rates using protein sequence-structure information there is no available method for predicting the unfolding rates of proteins upon specific point mutations. In this work, we have systematically analyzed a set of 790 single mutants and developed a robust method for predicting protein unfolding rates upon mutations (Δlnku) in two-state proteins by combining amino acid properties and knowledge-based classification of mutants with multiple linear regression technique. We obtain a mean absolute error (MAE) of 0.79/s and a Pearson correlation coefficient (PCC) of 0.71 between predicted unfolding rates and experimental observations using jack-knife test. We have developed a web server for predicting protein unfolding rates upon mutation and it is freely available at https://www.iitm.ac.in/bioinfo/proteinunfolding/unfoldingrace.html. Prominent features that determine unfolding kinetics as well as plausible reasons for the observed outliers are also discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

HPLC PROFILING OF PHENOLIC ACIDS AND FLAVONOIDS AND EVALUATION OF ANTI-LIPOXYGENASE AND ANTIOXIDANT ACTIVITIES OF AQUATIC VEGETABLE LIMNOCHARIS FLAVA.

PubMed

Ooh, Keng-fei; Ong, Hean-Chooi; Wong, Fai-Chu; Chai, Tsun-Thai

2015-01-01

Limnocharis flava is an edible wetland plant, whose phenolic acid and flavonoid compositions as well as bioactivities were underexplored. This study analyzed the profiles of selected hydroxybenzoic acids, hydroxycinnamic acids and flavonoids in the aqueous extracts of L. flava leaf, rhizome and root by high performance liquid chromatography (HPLC). Anti-lipoxygenase and antioxidant (iron chelating, 2,2-diphenyl-l-picrylhydrazyl (DPPH) radical scavenging, and nitric oxide (NO) scavenging) activities of the extracts were also evaluated. Leaf extract had the highest phenolic contents, being most abundant in p-hydroxybenzoic acid (3861.2 nmol/g dry matter), ferulic acid (648.8 nmol/g dry matter), and rutin (4110.7 nmol/g dry matter). Leaf extract exhibited the strongest anti-lipoxygenase (EC50 6.47 mg/mL), iron chelating (EC50 6.65 mg/mL), DPPH scavenging (EC50 15.82 mg/mL) and NO scavenging (EC50 3.80 mg/mL) activities. Leaf extract also had the highest ferric reducing ability. This is the most extensive HPLC profiling of phenolic acids and flavonoids in L.flava to date. In conclusion, L. flava leaf is a source of health-promoting phenolics, anti-lipoxygenase agents and antioxidants.

Compound-Specific Radiocarbon Dating Reveals the Age Distribution of Plant-Wax Biomarkers Exported to the Bengal Fan

NASA Astrophysics Data System (ADS)

Galy, V.; French, K. L.; Hein, C. J.; Haghipour, N.; Wacker, L.; Kudrass, H.; Eglinton, T. I.

2017-12-01

The stable isotope composition of leaf-wax compounds preserved in lacustrine and marine sediments has been widely used to reconstruct terrestrial paleo-environments. However, the timescales of plant-wax storage in continental reservoirs before riverine export are not well known, representing a key uncertainty in paleo-environment studies. We couple numerical models with bulk and leaf-wax fatty acid organic 13C and 14C signatures hosted in a high-deposition-rate sediment core from the Bengal shelf canyon in order to estimate storage timescales within the Ganges-Brahmaputra catchment area. The fatty acid 14C record reveals a muted nuclear weapons bomb spike, requiring that the Ganges-Brahmaputra river system exports a mixture of young and old (pre-aged) leaf-wax compounds. According to numerical simulations, 79-83% of the leaf-wax fatty acids in this core are sourced from continental reservoirs that store organic carbon on an average of 1000-1200 calendar years, while the remainder has an average age of 15 years. These results demonstrate that a majority of the leaf-wax compounds produced in the Ganges-Brahmaputra river basin was stored in soils, floodplains, and wetlands prior to its export to the Bengal Fan. We will discuss the implications of these findings for plant-wax based paleoenvironmental records.

Aerodynamic roughness: A simple and alternative metric to detect the seasonality of canopy structure using flux-tower data

NASA Astrophysics Data System (ADS)

Chu, H.; Baldocchi, D. D.

2017-12-01

FLUXNET - the global network of eddy covariance tower sites provides valuable datasets of the direct and in situ measurements of fluxes and ancillary variables that are used across different disciplines and applications. Aerodynamic roughness (i.e., roughness length, zero plane displacement height) are one of the potential parameters that can be derived from flux-tower data and are crucial for the applications of land surface models and flux footprint models. As aerodynamic roughness are tightly associated with canopy structures (e.g., canopy height, leaf area), such parameters could potentially serve as an alternative metric for detecting the change of canopy structure (e.g., change of leaf areas in deciduous ecosystems). This study proposes a simple approach for deriving aerodynamic roughness from flux-tower data, and tests their suitability and robustness in detecting the seasonality of canopy structure. We run tests across a broad range of deciduous forests, and compare the seasonality derived from aerodynamic roughness (i.e., starting and ending dates of leaf-on period and peak-foliage period) against those obtained from remote sensing or in situ leaf area measurements. Our findings show aerodynamic roughness generally captures the timing of changes of leaf areas in deciduous forests. Yet, caution needs to be exercised while interpreting the absolute values of the roughness estimates.

Tuning Transpiration by Interfacial Solar Absorber‐Leaf Engineering

PubMed Central

Zhuang, Shendong; Zhou, Lin; Xu, Weichao; Xu, Ning; Hu, Xiaozhen; Li, Xiuqiang; Lv, Guangxin; Zheng, Qinghui; Zhu, Shining

2017-01-01

Abstract Plant transpiration, a process of water movement through a plant and its evaporation from aerial parts especially leaves, consumes a large component of the total continental precipitation (≈48%) and significantly influences global water distribution and climate. To date, various chemical and/or biological explorations have been made to tune the transpiration but with uncertain environmental risks. In recent years, interfacial solar steam/vapor generation is attracting a lot of attention for achieving high energy transfer efficiency. Various optical and thermal designs at the solar absorber–water interface for potential applications in water purification, seawater desalination, and power generation appear. In this work, the concept of interfacial solar vapor generation is extended to tunable plant transpiration by showing for the first time that the transpiration efficiency can also be enhanced or suppressed through engineering the solar absorber–leaf interface. By tuning the solar absorption of membrane in direct touch with green leaf, surface temperature of green leaf will change accordingly because of photothermal effect, thus the transpiration efficiency as well as temperature and relative humidity in the surrounding environment will be tuned. This tunable transpiration by interfacial absorber‐leaf engineering can open an alternative avenue to regulate local atmospheric temperature, humidity, and eventually hydrologic cycle. PMID:29619300

Unfolding pathway of CotA-laccase and the role of copper on the prevention of refolding through aggregation of the unfolded state

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

Fernandes, Andre T.; Lopes, Carlos; Martins, Ligia O.

2012-06-08

Highlights: Black-Right-Pointing-Pointer CotA-laccase unfolds with an intermediate state. Black-Right-Pointing-Pointer Copper stabilizes the native and the intermediate state. Black-Right-Pointing-Pointer Copper binding to the unfolded state prevents refolding through protein aggregation. Black-Right-Pointing-Pointer Copper incorporation in CotA-laccase occurs as a later step during folding. -- Abstract: Copper is a redox-active metal and the main player in electron transfer reactions occurring in multicopper oxidases. The role of copper in the unfolding pathway and refolding of the multicopper oxidase CotA laccase in vitro was solved using double-jump stopped-flow experiments. Unfolding of apo- and holo-CotA was described as a three-state process with accumulation of an intermediatemore » in between the native and unfolded state. Copper stabilizes the native holo-CotA but also the intermediate state showing that copper is still bound to this state. Also, copper binds to unfolded holo-CotA in a non-native coordination promoting CotA aggregation and preventing refolding to the native structure. These results gather information on unfolding/folding pathways of multicopper oxidases and show that copper incorporation in vivo should be a tight controlled process as copper binding to the unfolded state under native conditions promotes protein aggregation.« less

Probing the contribution of internal cavities to the volume change of protein unfolding under pressure.

PubMed Central

Frye, K. J.; Royer, C. A.

1998-01-01

The structural origin of the decrease in system volume upon protein denaturation by pressure has remained a puzzle for decades. This negative volume change upon unfolding is assumed to arise globally from more intimate interactions between the polypeptide chain and water, including electrostriction of buried charges that become exposed upon unfolding, hydration of the polypeptide backbone and amino acid side chains and elimination of packing defects and internal void volumes upon unfolding of the chain. However, the relative signs and magnitudes of each of these contributing factors have not been experimentally determined. Our laboratory has probed the fundamental basis for the volume change upon unfolding of staphylococcal nuclease (Snase) using variable solution conditions and point mutants of Snase (Royer CA et al., 1993, Biochemistry 32:5222-5232; Frye KJ et al., 1996, Biochemistry 35:10234-10239). Our prior results indicate that for Snase, neither electrostriction nor polar or nonpolar hydration contributes significantly to the value of the volume change of unfolding. In the present work, we investigate the pressure induced unfolding of three point mutants of Snase in which internal cavity size is altered. The experimentally determined volume changes of unfolding for the mutants suggest that loss of internal void volume upon unfolding represents the major contributing factor to the value of the volume change of Snase unfolding. PMID:9792110

Beetle wings are inflatable origami

NASA Astrophysics Data System (ADS)

Chen, Rui; Ren, Jing; Ge, Siqin; Hu, David

2015-11-01

Beetles keep their wings folded and protected under a hard shell. In times of danger, they must unfold them rapidly in order for them to fly to escape. Moreover, they must do so across a range of body mass, from 1 mg to 10 grams. How can they unfold their wings so quickly? We use high-speed videography to record wing unfolding times, which we relate to the geometry of the network of blood vessels in the wing. Larger beetles have longer unfolding times. Modeling of the flow of blood through the veins successfully accounts for the wing unfolding speed of large beetles. However, smaller beetles have anomalously short unfolding times, suggesting they have lower blood viscosity or higher driving pressure. The use of hydraulics to unfold complex objects may have implications in the design of micro-flying air vehicles.

History, rare, and multiple events of mechanical unfolding of repeat proteins

NASA Astrophysics Data System (ADS)

Sumbul, Fidan; Marchesi, Arin; Rico, Felix

2018-03-01

Mechanical unfolding of proteins consisting of repeat domains is an excellent tool to obtain large statistics. Force spectroscopy experiments using atomic force microscopy on proteins presenting multiple domains have revealed that unfolding forces depend on the number of folded domains (history) and have reported intermediate states and rare events. However, the common use of unspecific attachment approaches to pull the protein of interest holds important limitations to study unfolding history and may lead to discarding rare and multiple probing events due to the presence of unspecific adhesion and uncertainty on the pulling site. Site-specific methods that have recently emerged minimize this uncertainty and would be excellent tools to probe unfolding history and rare events. However, detailed characterization of these approaches is required to identify their advantages and limitations. Here, we characterize a site-specific binding approach based on the ultrastable complex dockerin/cohesin III revealing its advantages and limitations to assess the unfolding history and to investigate rare and multiple events during the unfolding of repeated domains. We show that this approach is more robust, reproducible, and provides larger statistics than conventional unspecific methods. We show that the method is optimal to reveal the history of unfolding from the very first domain and to detect rare events, while being more limited to assess intermediate states. Finally, we quantify the forces required to unfold two molecules pulled in parallel, difficult when using unspecific approaches. The proposed method represents a step forward toward more reproducible measurements to probe protein unfolding history and opens the door to systematic probing of rare and multiple molecule unfolding mechanisms.

Folding and unfolding pathway of chaperonin GroEL monomer and elucidation of thermodynamic parameters.

PubMed

Puri, Sarita; Chaudhuri, Tapan K

2017-03-01

The conformation and thermodynamic stability of monomeric GroEL were studied by CD and fluorescence spectroscopy. GroEL denaturation with urea and dilution in buffer leads to formation of a folded GroEL monomer. The monomeric nature of this protein was verified by size-exclusion chromatography and native PAGE. It has a well-defined secondary and tertiary structure, folding activity (prevention of aggregation) for substrate protein and is resistant to proteolysis. Being a properly folded and reversibly refoldable, monomeric GroEL is amenable for the study of thermodynamic stability by unfolding transition methods. We present the equilibrium unfolding of monomeric GroEL as studied by urea and heat mediated unfolding processes. The urea mediated unfolding shows two transitions and a single transition in the heat mediated unfolding process. In the case of thermal unfolding, some residual structure unfolds at a higher temperature (70-75°C). The process of folding/unfolding is reversible in both cases. Analysis of folding/unfolding data provides a measure of ΔG NU H 2 O , T m , ΔH van and ΔS van of monomeric GroEL. The thermodynamic stability parameter ΔG NU H 2 O is similar with both CD and intrinsic fluorescence i.e. 7.10±1.0kcal/mol. The calculated T m , ΔH van and ΔS van from the thermal unfolding transition is 46±0.5°C, 43.3±0.1kcal/mol and 143.9±0.1cal/mol/k respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

Towards an understanding of coupled physical and biological processes in the cultivated Sahel - 2. Vegetation and carbon dynamics

NASA Astrophysics Data System (ADS)

Boulain, N.; Cappelaere, B.; Ramier, D.; Issoufou, H. B. A.; Halilou, O.; Seghieri, J.; Guillemin, F.; Oï, M.; Gignoux, J.; Timouk, F.

2009-08-01

SummaryThis paper analyses the dynamics of vegetation and carbon during the West African monsoon season, for millet crop and fallow vegetation covers in the cultivated area of the Sahel. Comparing these two dominant land cover types informs on the impact of cultivation on productivity and carbon fluxes. Biomass, leaf area index (LAI) and carbon fluxes were monitored over a 2-year period for these two vegetation systems in the Wankama catchment of the AMMA (African monsoon multidisciplinary analyses) experimental super-site in West Niger. Carbon fluxes and water use efficiency observed at the field scale are confronted with ecophysiological measurements (photosynthetic response to light, and relation of water use efficiency to air humidity) made at the leaf scale for the dominant plant species in the two vegetation systems. The two rainy seasons monitored were dissimilar with respect to rain patterns, reflecting some of the interannual variability. Distinct responses in vegetation development and in carbon dynamics were observed between the two vegetation systems. Vegetation development in the fallow was found to depend more on rainfall distribution along the season than on its starting date. A quite opposite behaviour was observed for the crop vegetation: the date of first rain appears as a principal factor of millet growth. Carbon flux exchanges were well correlated to vegetation development. High responses of photosynthesis to light were observed for the dominant herbaceous and shrub species of the fallow at the leaf and field scales. Millet showed high response at the leaf scale, but a much lesser response at the field scale. This pattern, also observed for water use efficiency, is to be related to the low density of the millet cover. A simple LAI-based model for scaling up the photosynthetic response from leaf to field scale was found quite successful for the fallow, but was less conclusive for the crop, due to spatial variability of LAI. Time/space variations in leaf distribution for the dominant species are key to scale transition of carbon dynamics. Results obtained for the two vegetation covers are important in light of the major land use/cover change experienced in the Sahel region due to extensive savanna clearing for food production.

The mechanochemistry of copper reports on the directionality of unfolding in model cupredoxin proteins

NASA Astrophysics Data System (ADS)

Beedle, Amy E. M.; Lezamiz, Ainhoa; Stirnemann, Guillaume; Garcia-Manyes, Sergi

2015-08-01

Understanding the directionality and sequence of protein unfolding is crucial to elucidate the underlying folding free energy landscape. An extra layer of complexity is added in metalloproteins, where a metal cofactor participates in the correct, functional fold of the protein. However, the precise mechanisms by which organometallic interactions are dynamically broken and reformed on (un)folding are largely unknown. Here we use single molecule force spectroscopy AFM combined with protein engineering and MD simulations to study the individual unfolding pathways of the blue-copper proteins azurin and plastocyanin. Using the nanomechanical properties of the native copper centre as a structurally embedded molecular reporter, we demonstrate that both proteins unfold via two independent, competing pathways. Our results provide experimental evidence of a novel kinetic partitioning scenario whereby the protein can stochastically unfold through two distinct main transition states placed at the N and C termini that dictate the direction in which unfolding occurs.

Complex Stability of Single Proteins Explored by Forced Unfolding Experiments

PubMed Central

Janovjak, Harald; Sapra, K. Tanuj; Müller, Daniel J.

2005-01-01

In the last decade atomic force microscopy has been used to measure the mechanical stability of single proteins. These force spectroscopy experiments have shown that many water-soluble and membrane proteins unfold via one or more intermediates. Recently, Li and co-workers found a linear correlation between the unfolding force of the native state and the intermediate in fibronectin, which they suggested indicated the presence of a molecular memory or multiple unfolding pathways (1). Here, we apply two independent methods in combination with Monte Carlo simulations to analyze the unfolding of α-helices E and D of bacteriorhodopsin (BR). We show that correlation analysis of unfolding forces is very sensitive to errors in force calibration of the instrument. In contrast, a comparison of relative forces provides a robust measure for the stability of unfolding intermediates. The proposed approach detects three energetically different states of α-helices E and D in trimeric BR. These states are not observed for monomeric BR and indicate that substantial information is hidden in forced unfolding experiments of single proteins. PMID:15792967

Complex stability of single proteins explored by forced unfolding experiments.

PubMed

Janovjak, Harald; Sapra, K Tanuj; Müller, Daniel J

2005-05-01

In the last decade atomic force microscopy has been used to measure the mechanical stability of single proteins. These force spectroscopy experiments have shown that many water-soluble and membrane proteins unfold via one or more intermediates. Recently, Li and co-workers found a linear correlation between the unfolding force of the native state and the intermediate in fibronectin, which they suggested indicated the presence of a molecular memory or multiple unfolding pathways (1). Here, we apply two independent methods in combination with Monte Carlo simulations to analyze the unfolding of alpha-helices E and D of bacteriorhodopsin (BR). We show that correlation analysis of unfolding forces is very sensitive to errors in force calibration of the instrument. In contrast, a comparison of relative forces provides a robust measure for the stability of unfolding intermediates. The proposed approach detects three energetically different states of alpha-helices E and D in trimeric BR. These states are not observed for monomeric BR and indicate that substantial information is hidden in forced unfolding experiments of single proteins.

Framework for Real-Time All-Hazards Global Situational Awareness

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

Omitaomu, Olufemi A; Fernandez, Steven J; Bhaduri, Budhendra L

Information systems play a pivotal role in emergency response by making consequence analysis models based on up-to-date data available to decision makers. While consequence analysis models have been used for years on local scales, their application on national and global scales has been constrained by lack of non-proprietary data. This chapter describes how this has changed using a framework for real-time all-hazards situational awareness called the Energy Awareness and Resiliency Standardized Services (EARSS) as an example. EARSS is a system of systems developed to collect non-proprietary data from diverse open content sources to develop a geodatabase of critical infrastructures allmore » over the world. The EARSS system shows that it is feasible to provide global disaster alerts by producing valuable information such as texting messages about detected hazards, emailing reports about affected areas, estimating an expected number of impacted people and their demographic characteristics, identifying critical infrastructures that may be affected, and analyzing potential downstream effects. This information is provided in real-time to federal agencies and subscribers all over the world for decision making in humanitarian assistance and emergency response. The system also uses live streams of power outages, weather, and satellite surveillance data as events unfold. This, in turn, is combined with other public domain or open content information, such as media reports and postings on social networking websites, for complete coverage of the situation as events unfold. Working with up-to-date information from the EARSS system, emergency responders on the ground could pre-position their staff and resources, such as emergency generators and ice, where they are most needed.« less

Spectroscopic Studies on Unfolding Processes of Apo-Neuroglobin Induced by Guanidine Hydrochloride and Urea

PubMed Central

Zhang, Cui; Gao, Chaohui; Qiu, Zhanglei

2013-01-01

Neuroglobin (Ngb), a recently discovered globin, is predominantly expressed in the brain, retina, and other nerve tissues of vertebrates. The unfolding processes of apo-neuroglobin (apoNgb) induced by guanidine hydrochloride (GdnHCl) and urea were investigated by spectroscopic methods. In the unfolding processes, apoNgb's tertiary structural transition was monitored by the changes of intrinsic fluorescence emission spectra, and its secondary structural transition was measured by the changes of far-ultraviolet circular dichroism (CD) spectra. In addition, 8-anilino-1-naphthalenesulfonic acid (ANS), a hydrophobic cluster binding dye, was also used to monitor the unfolding process of apoNgb and to explore its intermediates. Results showed that GdnHCl-induced unfolding of apoNgb was via a three-state pathway, that is, Native state (N) → Intermediate state (I) → Unfolded state (U), during which the intermediate was inferred by an increase in fluorescence intensity and the change of CD value. Gibbs free energy changes are 10.2 kJ·mol−1 for the first unfolding transition and 14.0 kJ·mol−1 for the second transition. However, urea-induced unfolding of apoNgb only underwent a two-state transition: Native state (N) → Partially unfolded state (P). The result showed that GdnHCl can efficiently affect the conformational states of apoNgb compared with those of urea. The work will benefit to have an understanding of the unfolding mechanism of apoNgb induced by GdnHCl and urea. PMID:23984347

Chemically crosslinked protein dimers: stability and denaturation effects.

PubMed Central

Byrne, M. P.; Stites, W. E.

1995-01-01

Nine single substitution cysteine mutants of staphylococcal nuclease (nuclease) were preferentially crosslinked at the introduced cysteine residues using three different bifunctional crosslinking reagents; 1,6-bismaleimidohexane (BMH), 1,3-dibromo-2-propanol (DBP), and the chemical warfare agent, mustard gas (bis(2-chloroethyl)sulfide; mustard). BMH and mustard gas are highly specific reagents for cysteine residues, whereas DBP is not as specific. Guanidine hydrochloride (GuHCl) denaturations of the resulting dimeric proteins exhibited biphasic unfolding behavior that did not fit the two-state model of unfolding. The monofunctional reagent, epsilon-maleimidocaproic acid (MCA), was used as a control for the effects of alkylation. Proteins modified with MCA unfolded normally, showing that this unusual unfolding behavior is due to crosslinking. The data obtained from these crosslinked dimers was fitted to a three-state thermodynamic model of two successive transitions in which the individual subunits cooperatively unfold. These two unfolding transitions were very different from the unfolding of the monomeric protein. These differences in unfolding behavior can be attributed in large part to changes in the denatured state. In addition to GuHCl titrations, the crosslinked dimers were also thermally unfolded. In contrast to the GuHCl denaturations, analysis of this data fit a two-state model well, but with greatly elevated van't Hoff enthalpies in many cases. However, clear correlations between the thermal and GuHCl denaturations exist, and the differences in thermal unfolding can be rationalized by postulating interactions of the denatured crosslinked proteins. PMID:8580845

The unfolding mechanism of monomeric mutant SOD1 by simulated force spectroscopy.

PubMed

Habibi, Mona; Rottler, Jörg; Plotkin, Steven S

2017-11-01

Mechanical unfolding of mutated apo, disulfide-reduced, monomeric superoxide dismutase 1 protein (SOD1) has been simulated via force spectroscopy techniques, using both an all-atom (AA), explicit solvent model and a coarse-grained heavy-atom Gō (HA-Gō) model. The HA-Gō model was implemented at two different pulling speeds for comparison. The most-common sequence of unfolding in the AA model agrees well with the most-common unfolding sequence of the HA-Gō model, when the same normalized pulling rate was used. Clustering of partially-native structures as the protein unfolds shows that the AA and HA-Gō models both exhibit a dominant pathway for early unfolding, which eventually bifurcates repeatedly to multiple branches after the protein is about half-unfolded. The force-extension curve exhibits multiple force drops, which are concomitant with jumps in the local interaction potential energy between specific β-strands in the protein. These sudden jumps in the potential energy coincide with the dissociation of specific pairs of β-strands, and thus intermediate unfolding events. The most common sequence of β-strand dissociation in the unfolding pathway of the AA model is β-strands 5, 4, 8, 7, 1, 2, then finally β-strands 3 and 6. The observation that β-strand 5 is among the first to unfold here, but the last to unfold in simulations of loop-truncated SOD1, could imply the existence of an evolutionary compensation mechanism, which would stabilize β-strands flanking long loops against their entropic penalty by strengthening intramolecular interactions. This article is part of a Special Issue entitled: Biophysics in Canada, edited by Lewis Kay, John Baenziger, Albert Berghuis and Peter Tieleman. Copyright © 2017 Elsevier B.V. All rights reserved.

Chemical unfolding of chicken villin headpiece in aqueous dimethyl sulfoxide solution: cosolvent concentration dependence, pathway, and microscopic mechanism.

PubMed

Roy, Susmita; Bagchi, Biman

2013-04-25

Unfolding of a protein often proceeds through partial unfolded intermediate states (PUIS). PUIS have been detected in several experimental and simulation studies. However, complete analyses of transitions between different PUIS and the unfolding trajectory are sparse. To understand such dynamical processes, we study chemical unfolding of a small protein, chicken villin head piece (HP-36), in aqueous dimethyl sulfoxide (DMSO) solution. We carry out molecular dynamics simulations at various solution compositions under ambient conditions. In each concentration, the initial step of unfolding involves separation of two adjacent native contacts, between phenyl alanine residues (11-18 and 7-18). This first step induces, under appropriate conditions, subsequent separation among other hydrophobic contacts, signifying a high degree of cooperativity in the unfolding process. The observed sequence of structural changes in HP-36 on increasing DMSO concentration and the observed sequence of PUIS, are in approximate agreement with earlier simulation results (in pure water) and experimental observations on unfolding of HP-36. Peculiar to water-DMSO mixture, an intervening structural transformation (around 15% of DMSO) in the binary mixture solvent retards the progression of unfolding as composition is increased. This is reflected in a remarkable nonmonotonic composition dependence of RMSD, radius of gyration and the fraction of native contacts. At 30% mole fraction of DMSO, we find the extended randomly coiled structure of the unfolded protein. The molecular mechanism of DMSO induced unfolding process is attributed to the initial preferential solvation of the hydrophobic side chain atoms through the methyl groups of DMSO, followed by the hydrogen bonding of the oxygen atom of DMSO to the exposed backbone NH groups of HP-36.

Unfolding study of a trimeric membrane protein AcrB.

PubMed

Ye, Cui; Wang, Zhaoshuai; Lu, Wei; Wei, Yinan

2014-07-01

The folding of a multi-domain trimeric α-helical membrane protein, Escherichia coli inner membrane protein AcrB, was investigated. AcrB contains both a transmembrane domain and a large periplasmic domain. Protein unfolding in sodium dodecyl sulfate (SDS) and urea was monitored using the intrinsic fluorescence and circular dichroism spectroscopy. The SDS denaturation curve displayed a sigmoidal profile, which could be fitted with a two-state unfolding model. To investigate the unfolding of separate domains, a triple mutant was created, in which all three Trp residues in the transmembrane domain were replaced with Phe. The SDS unfolding profile of the mutant was comparable to that of the wild type AcrB, suggesting that the observed signal change was largely originated from the unfolding of the soluble domain. Strengthening of trimer association through the introduction of an inter-subunit disulfide bond had little effect on the unfolding profile, suggesting that trimer dissociation was not the rate-limiting step in unfolding monitored by fluorescence emission. Under our experimental condition, AcrB unfolding was not reversible. Furthermore, we experimented with the refolding of a monomeric mutant, AcrBΔloop , from the SDS unfolded state. The CD spectrum of the refolded AcrBΔloop superimposed well onto the spectra of the original folded protein, while the fluorescence spectrum was not fully recovered. In summary, our results suggested that the unfolding of the trimeric AcrB started with a local structural rearrangement. While the refolding of secondary structure in individual monomers could be achieved, the re-association of the trimer might be the limiting factor to obtain folded wild-type AcrB. © 2014 The Protein Society.

Studying pressure denaturation of a protein by molecular dynamics simulations.

PubMed

Sarupria, Sapna; Ghosh, Tuhin; García, Angel E; Garde, Shekhar

2010-05-15

Many globular proteins unfold when subjected to several kilobars of hydrostatic pressure. This "unfolding-up-on-squeezing" is counter-intuitive in that one expects mechanical compression of proteins with increasing pressure. Molecular simulations have the potential to provide fundamental understanding of pressure effects on proteins. However, the slow kinetics of unfolding, especially at high pressures, eliminates the possibility of its direct observation by molecular dynamics (MD) simulations. Motivated by experimental results-that pressure denatured states are water-swollen, and theoretical results-that water transfer into hydrophobic contacts becomes favorable with increasing pressure, we employ a water insertion method to generate unfolded states of the protein Staphylococcal Nuclease (Snase). Structural characteristics of these unfolded states-their water-swollen nature, retention of secondary structure, and overall compactness-mimic those observed in experiments. Using conformations of folded and unfolded states, we calculate their partial molar volumes in MD simulations and estimate the pressure-dependent free energy of unfolding. The volume of unfolding of Snase is negative (approximately -60 mL/mol at 1 bar) and is relatively insensitive to pressure, leading to its unfolding in the pressure range of 1500-2000 bars. Interestingly, once the protein is sufficiently water swollen, the partial molar volume of the protein appears to be insensitive to further conformational expansion or unfolding. Specifically, water-swollen structures with relatively low radii of gyration have partial molar volume that are similar to that of significantly more unfolded states. We find that the compressibility change on unfolding is negligible, consistent with experiments. We also analyze hydration shell fluctuations to comment on the hydration contributions to protein compressibility. Our study demonstrates the utility of molecular simulations in estimating volumetric properties and pressure stability of proteins, and can be potentially extended for applications to protein complexes and assemblies. Proteins 2010. (c) 2009 Wiley-Liss, Inc.

Leaf Tissue C:N and Soil N are Modified by Growing Season and Goose Grazing Phenology in a Sub-Arctic Coastal Wetland of Western Alaska

NASA Astrophysics Data System (ADS)

Choi, R. T.; Beard, K. H.; Leffler, A. J.; Schmutz, J. A.; Welker, J. M.

2014-12-01

Climate change in Arctic wetlands is resulting in a widening phenological mismatch between the onset of the growing season and the arrival and hatch date of migratory geese, the primary consumers in the system. During the past three decades, the growing season has advanced but geese have not advanced arrival or hatch date at the same rate. Geese now arrive into a system that has been growing longer than in the past with potential changes in forage quality because sedges have their highest nutrient density shortly following emergence. One potential concomitant result of this phenological gap is altered carbon to nitrogen ratio (C:N) of leaf tissue being returned to the ecosystem as feces that is more N-poor. Altering the C:N of these inputs can further influence C and N cycling in the system. We examine the influence of advanced growing season and different arrival times by black brant on leaf and soil C:N ratio and soil N-form. Our experiment consists of six blocks with nine study plots each. Half the plots are warmed to advance the growing season. Two plots each receive early, typical, late, and no grazing; one plot is a control that is not warmed and grazing is natural. Leaf tissue was collected to determine C and N concentration using an elemental analyzer. Anion and cation exchange membranes were used to monitor inorganic N forms in soil; samples were analyzed via fluorescence following extraction. Soil water collected from lysimeters was analyzed for organic N. Warming advanced plant growth between one and two weeks and resulted in higher C:N of leaf tissue Geese maintained 'grazing lawns', areas of exceptionally short vegetation, where plants had high N compared to non-grazed areas. Grazing early in the season promoted higher N content of leaves and soil while grazing late had little influence on N. The timing of the growing season and grazing both have important implications for C and N in this system.

Arabidopsis Thaliana Ecotypes With Differential Susceptibility To The Bacterial Pathogen Xylella fastidiosa

USDA-ARS?s Scientific Manuscript database

Pierce’s disease of grapes and almond leaf scorch are devastating diseases caused by the bacterium Xylella fastidiosa (Xf). To date, progress in determining the mechanisms of host plant susceptibility, tolerance or resistance has been slow, due in large part to the long generation time and limited a...

Effect of understory management on phenological responses of eastern black walnut on an alluvial Arkansas soil

USDA-ARS?s Scientific Manuscript database

Black walnut (Juglans nigra L.) is commonly grown in agroforestry practices for nuts and/or timber with little knowledge of how understory herbage management might affect tree phenology. We compared black walnut plant type (variety and wild-type) for phenological response in date of budburst, leaf ...

Effects of ambient ozone on first-year growth and physiology of black cherry (Prunus serotina Ehrh. ) seedlings

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

Kolb, T.E.; Skelly, J.M.; Steiner, K.C.

1993-06-01

Black cherry seedlings of two open-pollinated families were exposed to ambient ozone in open plots (100%) and plots receiving non-filtered (95%), half-filtered (60%) and full-filtered (40%) concentrations via open-top chambers between early June and late-September 1992 in a replicated field experiment in central Pennsylvania. Seasonal 24-hour ambient ozone concentration averaged 34 ppb with a peak 1-hour concentration of 110 ppb. Foliar symptoms of ozone damage (adaxial stipple) occurred most prominently in open and non-filtered plots and differed between families. Net photosynthetic rate for both families was significantly lower in open and non-filtered plots compared with half- and full-filtered plots onmore » most dates, while ozone concentration had no consistent effect on leaf conductance or dark respiration. Leaf conductance of the ozone sensitive family was significantly greater than the ozone tolerant family on most dates. First-year height and diameter growth were significantly lower in open and non-filtered plots compared with half- and full-filtered plots for both families.« less

Effect of temperature on the conformation of natively unfolded protein 4E-BP1 in aqueous and mixed solutions containing trifluoroethanol and hexafluoroisopropanol.

PubMed

Hackl, Ellen V

2015-02-01

Natively unfolded (intrinsically disordered) proteins have attracted growing attention due to their high abundance in nature, involvement in various signalling and regulatory pathways and direct association with many diseases. In the present work the combined effect of temperature and alcohols, trifluoroethanol (TFE) and hexafluoroisopropanol (HFIP), on the natively unfolded 4E-BP1 protein was studied to elucidate the balance between temperature-induced folding and unfolding in intrinsically disordered proteins. It was shown that elevated temperatures induce reversible partial folding of 4E-BP1 both in buffer and in the mixed solutions containing denaturants. In the mixed solutions containing TFE (HFIP) 4E-BP1 adopts a partially folded helical conformation. As the temperature increases, the initial temperature-induced protein folding is replaced by irreversible unfolding/melting only after a certain level of the protein helicity has been reached. Onset unfolding temperature decreases with TFE (HFIP) concentration in solution. It was shown that an increase in the temperature induces two divergent processes in a natively unfolded protein--hydrophobicity-driven folding and unfolding. Balance between these two processes determines thermal behaviour of a protein. The correlation between heat-induced protein unfolding and the amount of helical content in a protein is revealed. Heat-induced secondary structure formation can be a valuable test to characterise minor changes in the conformations of natively unfolded proteins as a result of site-directed mutagenesis. Mutants with an increased propensity to fold into a structured form reveal different temperature behaviour.

Unfolding of the cold shock protein studied with biased molecular dynamics.

PubMed

Morra, Giulia; Hodoscek, Milan; Knapp, Ernst-Walter

2003-11-15

The cold shock protein from Bacillus caldolyticus is a small beta-barrel protein that folds in a two-state mechanism. For the native protein and for several mutants, a wealth of experimental data are available on stability and folding, so that it is an optimal system to study this process. We compare data from unfolding simulations (trajectories of 5 and up to 12 ns) obtained with a bias potential at room temperature and from unbiased thermal unfolding simulations with experimental data. The unfolding patterns derived from the trajectories starting from different native-like conformations and subject to different unfolding conditions agree. The transition state found in the simulations of unfolding is close to the native structure in agreement with experiment. Moreover, a lower value of the free energy barrier of unfolding was found for the mutant R3E than for the mutant E46A and the native protein, as indicated by experimental data. The first unfolding event involves the three-stranded beta-sheet whose decomposition corresponds to the transition state. In contrast to conclusions drawn from experiments, we found that the two-stranded beta-strand forms the most stable substructure, which decomposes very late in the unfolding process. However, assuming that this structure forms very early in the folding process, our findings would not contradict the experiments but require a different interpretation of them. Copyright 2003 Wiley-Liss, Inc.

Application of long-range order to predict unfolding rates of two-state proteins.

PubMed

Harihar, B; Selvaraj, S

2011-03-01

Predicting the experimental unfolding rates of two-state proteins and models describing the unfolding rates of these proteins is quite limited because of the complexity present in the unfolding mechanism and the lack of experimental unfolding data compared with folding data. In this work, 25 two-state proteins characterized by Maxwell et al. (Protein Sci 2005;14:602–616) using a consensus set of experimental conditions were taken, and the parameter long-range order (LRO) derived from their three-dimensional structures were related with their experimental unfolding rates ln(k(u)). From the total data set of 30 proteins used by Maxwell et al. (Protein Sci 2005;14:602–616), five slow-unfolding proteins with very low unfolding rates were considered to be outliers and were not included in our data set. Except all beta structural class, LRO of both the all-alpha and mixed-class proteins showed a strong inverse correlation of r = -0.99 and -0.88, respectively, with experimental ln(k(u)). LRO shows a correlation of -0.62 with experimental ln(k(u)) for all-beta proteins. For predicting the unfolding rates, a simple statistical method has been used and linear regression equations were developed for individual structural classes of proteins using LRO, and the results obtained showed a better agreement with experimental results. Copyright © 2010 Wiley-Liss, Inc.

Deconvoluting Protein (Un)folding Structural Ensembles Using X-Ray Scattering, Nuclear Magnetic Resonance Spectroscopy and Molecular Dynamics Simulation

PubMed Central

Nasedkin, Alexandr; Marcellini, Moreno; Religa, Tomasz L.; Freund, Stefan M.; Menzel, Andreas; Fersht, Alan R.; Jemth, Per; van der Spoel, David; Davidsson, Jan

2015-01-01

The folding and unfolding of protein domains is an apparently cooperative process, but transient intermediates have been detected in some cases. Such (un)folding intermediates are challenging to investigate structurally as they are typically not long-lived and their role in the (un)folding reaction has often been questioned. One of the most well studied (un)folding pathways is that of Drosophila melanogaster Engrailed homeodomain (EnHD): this 61-residue protein forms a three helix bundle in the native state and folds via a helical intermediate. Here we used molecular dynamics simulations to derive sample conformations of EnHD in the native, intermediate, and unfolded states and selected the relevant structural clusters by comparing to small/wide angle X-ray scattering data at four different temperatures. The results are corroborated using residual dipolar couplings determined by NMR spectroscopy. Our results agree well with the previously proposed (un)folding pathway. However, they also suggest that the fully unfolded state is present at a low fraction throughout the investigated temperature interval, and that the (un)folding intermediate is highly populated at the thermal midpoint in line with the view that this intermediate can be regarded to be the denatured state under physiological conditions. Further, the combination of ensemble structural techniques with MD allows for determination of structures and populations of multiple interconverting structures in solution. PMID:25946337

Deconvoluting Protein (Un)folding Structural Ensembles Using X-Ray Scattering, Nuclear Magnetic Resonance Spectroscopy and Molecular Dynamics Simulation.

PubMed

Nasedkin, Alexandr; Marcellini, Moreno; Religa, Tomasz L; Freund, Stefan M; Menzel, Andreas; Fersht, Alan R; Jemth, Per; van der Spoel, David; Davidsson, Jan

2015-01-01

The folding and unfolding of protein domains is an apparently cooperative process, but transient intermediates have been detected in some cases. Such (un)folding intermediates are challenging to investigate structurally as they are typically not long-lived and their role in the (un)folding reaction has often been questioned. One of the most well studied (un)folding pathways is that of Drosophila melanogaster Engrailed homeodomain (EnHD): this 61-residue protein forms a three helix bundle in the native state and folds via a helical intermediate. Here we used molecular dynamics simulations to derive sample conformations of EnHD in the native, intermediate, and unfolded states and selected the relevant structural clusters by comparing to small/wide angle X-ray scattering data at four different temperatures. The results are corroborated using residual dipolar couplings determined by NMR spectroscopy. Our results agree well with the previously proposed (un)folding pathway. However, they also suggest that the fully unfolded state is present at a low fraction throughout the investigated temperature interval, and that the (un)folding intermediate is highly populated at the thermal midpoint in line with the view that this intermediate can be regarded to be the denatured state under physiological conditions. Further, the combination of ensemble structural techniques with MD allows for determination of structures and populations of multiple interconverting structures in solution.

Folding thermodynamics of pseudoknotted chain conformations

PubMed Central

Kopeikin, Zoia; Chen, Shi-Jie

2008-01-01

We develop a statistical mechanical framework for the folding thermodynamics of pseudoknotted structures. As applications of the theory, we investigate the folding stability and the free energy landscapes for both the thermal and the mechanical unfolding of pseudoknotted chains. For the mechanical unfolding process, we predict the force-extension curves, from which we can obtain the information about structural transitions in the unfolding process. In general, a pseudoknotted structure unfolds through multiple structural transitions. The interplay between the helix stems and the loops plays an important role in the folding stability of pseudoknots. For instance, variations in loop sizes can lead to the destabilization of some intermediate states and change the (equilibrium) folding pathways (e.g., two helix stems unfold either cooperatively or sequentially). In both thermal and mechanical unfolding, depending on the nucleotide sequence, misfolded intermediate states can emerge in the folding process. In addition, thermal and mechanical unfoldings often have different (equilibrium) pathways. For example, for certain sequences, the misfolded intermediates, which generally have longer tails, can fold, unfold, and refold again in the pulling process, which means that these intermediates can switch between two different average end-end extensions. PMID:16674261

Chemical probes and engineered constructs reveal a detailed unfolding mechanism for a solvent-free multi-domain protein

PubMed Central

Eschweiler, Joseph D.; Martini, Rachel M.; Ruotolo, Brandon T.

2017-01-01

Despite the growing application of gas-phase measurements in structural biology and drug discovery, the factors that govern protein stabilities and structures in a solvent-free environment are still poorly understood. Here, we examine the solvent-free unfolding pathway for a group of homologous serum albumins. Utilizing a combination of chemical probes and non-covalent reconstructions, we draw new specific conclusions regarding the unfolding of albumins in the gas-phase, as well as more-general inferences regarding the sensitivity of collision induced unfolding to changes in protein primary and tertiary structure. Our findings suggest that the general unfolding pathway of low charge state albumin ions is largely unaffected by changes in primary structure; however, the stabilities of intermediates along these pathways vary widely as sequences diverge. Additionally, we find that human albumin follows a domain associated unfolding pathway, and are able to assign each unfolded form observed in our gas-phase dataset to the disruption of specific domains within the protein. The totality of our data informs the first detailed mechanism for multi-domain protein unfolding in the gas phase, and highlights key similarities and differences from the known the solution-phase pathway. PMID:27959526

Comparative study of protein unfolding in aqueous urea and dimethyl sulfoxide solutions: surface polarity, solvent specificity, and sequence of secondary structure melting.

PubMed

Roy, Susmita; Bagchi, Biman

2014-05-29

Elucidation of possible pathways between folded (native) and unfolded states of a protein is a challenging task, as the intermediates are often hard to detect. Here, we alter the solvent environment in a controlled manner by choosing two different cosolvents of water, urea, and dimethyl sulfoxide (DMSO) and study unfolding of four different proteins to understand the respective sequence of melting by computer simulation methods. We indeed find interesting differences in the sequence of melting of α helices and β sheets in these two solvents. For example, in 8 M urea solution, β-sheet parts of a protein are found to unfold preferentially, followed by the unfolding of α helices. In contrast, 8 M DMSO solution unfolds α helices first, followed by the separation of β sheets for the majority of proteins. Sequence of unfolding events in four different α/β proteins and also in chicken villin head piece (HP-36) both in urea and DMSO solutions demonstrate that the unfolding pathways are determined jointly by relative exposure of polar and nonpolar residues of a protein and the mode of molecular action of a solvent on that protein.

Impact of ion binding on poly-L-lysine (un)folding energy landscape and kinetics.

PubMed

Xiong, Kan; Asher, Sanford A

2012-06-21

We utilize T-jump UV resonance Raman spectroscopy (UVRR) to study the impact of ion binding on the equilibrium energy landscape and on (un)folding kinetics of poly-L-lysine (PLL). We observe that the relaxation rates of the folded conformations (including π-helix (bulge), pure α-helix, and turns) of PLL are slower than those of short alanine-based peptides. The PLL pure α-helix folding time is similar to that of short alanine-based peptides. We for the first time have directly observed that turn conformations are α-helix and π-helix (bulge) unfolding intermediates. ClO(4)(-) binding to the Lys side chain -NH(3)(+) groups and the peptide backbone slows the α-helix unfolding rate compared to that in pure water, but little impacts the folding rate, resulting in an increased α-helix stability. ClO(4)(-) binding significantly increases the PLL unfolding activation barrier but little impacts the folding barrier. Thus, the PLL folding coordinate(s) differs from the unfolding coordinate(s). The-π helix (bulge) unfolding and folding coordinates do not directly go through the α-helix energy well. Our results clearly demonstrate that PLL (un)folding is not a two-state process.

Distinct transcriptional responses elicited by unfolded nuclear or cytoplasmic protein in mammalian cells

PubMed Central

Miyazaki, Yusuke; Chen, Ling-chun; Chu, Bernard W; Swigut, Tomek; Wandless, Thomas J

2015-01-01

Eukaryotic cells possess a variety of signaling pathways that prevent accumulation of unfolded and misfolded proteins. Chief among these is the heat shock response (HSR), which is assumed to respond to unfolded proteins in the cytosol and nucleus alike. In this study, we probe this axiom further using engineered proteins called ‘destabilizing domains’, whose folding state we control with a small molecule. The sudden appearance of unfolded protein in mammalian cells elicits a robust transcriptional response, which is distinct from the HSR and other known pathways that respond to unfolded proteins. The cellular response to unfolded protein is strikingly different in the nucleus and the cytosol, although unfolded protein in either compartment engages the p53 network. This response provides cross-protection during subsequent proteotoxic stress, suggesting that it is a central component of protein quality control networks, and like the HSR, is likely to influence the initiation and progression of human pathologies. DOI: http://dx.doi.org/10.7554/eLife.07687.001 PMID:26314864

A method to determine residue-specific unfolded-state pKa values from analysis of stability changes in single mutant cycles.

PubMed

Shen, Jana K

2010-06-02

It is now widely recognized that the unfolded state of a protein in equilibrium with the native state under folding conditions may contain significant residual structures. However, due to technical difficulties residue-specific interactions in the unfolded state remain elusive. Here we introduce a method derived from the Wyman-Tanford theory to determine residue-specific pK(a)'s in the unfolded state. This method requires equilibrium stability measurements of the wild type and single-point mutants in which titrable residues are replaced with charge-neutral ones under two pH conditions. Application of the proposed approach reveals a highly depressed pK(a) for Asp8 in the unfolded state of the NTL9 protein. Knowledge of unfolded-state pK(a)'s enables quantitative estimation of the unfolded-state electrostatic effects on protein stability. It also provides valuable benchmarks for the improvement of force fields and validation of microscopic information from molecular dynamics simulations.

High temperature unfolding of a truncated hemoglobin by molecular dynamics simulation.

PubMed

Sharma, Ravi Datta; Kanwal, Rajnee; Lynn, Andrew M; Singh, Prerna; Pasha, Syed Tazeen; Fatma, Tasneem; Jawaid, Safdar

2013-09-01

Heme containing proteins are associated with peroxidase activity. The proteins like hemoglobin, myoglobins, cytochrome c and micro-peroxidase other than peroxidases have been shown to exhibit weak peroxidase-like activity. This weak peroxidase-like activity in hemoglobin-like molecules is due to heme moiety. We conducted molecular dynamics (MD) studies to decipher the unfolding path of Ba-Glb (a truncated hemoglobin from Bacillus anthracis) and the role of heme moiety to its unfolding path. The similar unfolding path is also observed in vitro by UV/VIS spectroscopy. The data confirmed that the unfolding of Ba-Glb follows a three state process with a meta-stable (intermediate) state between the native and unfolded conformations. The present study is supported by several unfolding parameters like root-mean-square-deviation (RMSD), dictionary of protein secondary structure (DSSP), and free energy landscape. Understanding the structure of hemoglobin like proteins in unicellular dreaded pathogens like B. anthracis will pave way for newer drug discovery targets and in the disease management of anthrax.

From observations to experiments in phenology research: investigating climate change impacts on trees and shrubs using dormant twigs.

PubMed

Primack, Richard B; Laube, Julia; Gallinat, Amanda S; Menzel, Annette

2015-11-01

Climate change is advancing the leaf-out times of many plant species and mostly extending the growing season in temperate ecosystems. Laboratory experiments using twig cuttings from woody plant species present an affordable, easily replicated approach to investigate the relative importance of factors such as winter chilling, photoperiod, spring warming and frost tolerance on the leafing-out times of plant communities. This Viewpoint article demonstrates how the results of these experiments deepen our understanding beyond what is possible via analyses of remote sensing and field observation data, and can be used to improve climate change forecasts of shifts in phenology, ecosystem processes and ecological interactions. The twig method involves cutting dormant twigs from trees, shrubs and vines on a single date or at intervals over the course of the winter and early spring, placing them in containers of water in controlled environments, and regularly recording leaf-out, flowering or other phenomena. Prior to or following leaf-out or flowering, twigs may be assigned to treatment groups for experiments involving temperature, photoperiod, frost, humidity and more. Recent studies using these methods have shown that winter chilling requirements and spring warming strongly affect leaf-out and flowering times of temperate trees and shrubs, whereas photoperiod requirements are less important than previously thought for most species. Invasive plant species have weaker winter chilling requirements than native species in temperate ecosystems, and species that leaf-out early in the season have greater frost tolerance than later leafing species. This methodology could be extended to investigate additional drivers of leaf-out phenology, leaf senescence in the autumn, and other phenomena, and could be a useful tool for education and outreach. Additional ecosystems, such as boreal, southern hemisphere and sub-tropical forests, could also be investigated using dormant twigs to determine the drivers of leaf-out times and how these ecosystems will be affected by climate change. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Dynamic quantification of canopy structure to characterize early plant vigour in wheat genotypes

PubMed Central

Duan, T.; Chapman, S.C.; Holland, E.; Rebetzke, G.J.; Guo, Y.; Zheng, B.

2016-01-01

Early vigour is an important physiological trait to improve establishment, water-use efficiency, and grain yield for wheat. Phenotyping large numbers of lines is challenging due to the fast growth and development of wheat seedlings. Here we developed a new photo-based workflow to monitor dynamically the growth and development of the wheat canopy of two wheat lines with a contrasting early vigour trait. Multiview images were taken using a ‘vegetation stress’ camera at 2 d intervals from emergence to the sixth leaf stage. Point clouds were extracted using the Multi-View Stereo and Structure From Motion (MVS-SFM) algorithm, and segmented into individual organs using the Octree method, with leaf midribs fitted using local polynomial function. Finally, phenotypic parameters were calculated from the reconstructed point cloud including: tiller and leaf number, plant height, Haun index, phyllochron, leaf length, angle, and leaf elongation rate. There was good agreement between the observed and estimated leaf length (RMSE=8.6mm, R 2=0.98, n=322) across both lines. Significant contrasts of phenotyping parameters were observed between the two lines and were consistent with manual observations. The early vigour line had fewer tillers (2.4±0.6) and larger leaves (308.0±38.4mm and 17.1±2.7mm for leaf length and width, respectively). While the phyllochron of both lines was quite similar, the non-vigorous line had a greater Haun index (more leaves on the main stem) on any date, as the vigorous line had slower development of its first two leaves. The workflow presented in this study provides an efficient method to phenotype individual plants using a low-cost camera (an RGB camera is also suitable) and could be applied in phenotyping for applications in both simulation modelling and breeding. The rapidity and accuracy of this novel method can characterize the results of specific selection criteria (e.g. width of leaf three, number of tillers, rate of leaf appearance) that have been or can now be utilized to breed for early leaf growth and tillering in wheat. PMID:27312669

Dynamic quantification of canopy structure to characterize early plant vigour in wheat genotypes.

PubMed

Duan, T; Chapman, S C; Holland, E; Rebetzke, G J; Guo, Y; Zheng, B

2016-08-01

Early vigour is an important physiological trait to improve establishment, water-use efficiency, and grain yield for wheat. Phenotyping large numbers of lines is challenging due to the fast growth and development of wheat seedlings. Here we developed a new photo-based workflow to monitor dynamically the growth and development of the wheat canopy of two wheat lines with a contrasting early vigour trait. Multiview images were taken using a 'vegetation stress' camera at 2 d intervals from emergence to the sixth leaf stage. Point clouds were extracted using the Multi-View Stereo and Structure From Motion (MVS-SFM) algorithm, and segmented into individual organs using the Octree method, with leaf midribs fitted using local polynomial function. Finally, phenotypic parameters were calculated from the reconstructed point cloud including: tiller and leaf number, plant height, Haun index, phyllochron, leaf length, angle, and leaf elongation rate. There was good agreement between the observed and estimated leaf length (RMSE=8.6mm, R (2)=0.98, n=322) across both lines. Significant contrasts of phenotyping parameters were observed between the two lines and were consistent with manual observations. The early vigour line had fewer tillers (2.4±0.6) and larger leaves (308.0±38.4mm and 17.1±2.7mm for leaf length and width, respectively). While the phyllochron of both lines was quite similar, the non-vigorous line had a greater Haun index (more leaves on the main stem) on any date, as the vigorous line had slower development of its first two leaves. The workflow presented in this study provides an efficient method to phenotype individual plants using a low-cost camera (an RGB camera is also suitable) and could be applied in phenotyping for applications in both simulation modelling and breeding. The rapidity and accuracy of this novel method can characterize the results of specific selection criteria (e.g. width of leaf three, number of tillers, rate of leaf appearance) that have been or can now be utilized to breed for early leaf growth and tillering in wheat. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Resistance of Brachystegia spiciformis to Carbohydrate and Phenological Manipulation

NASA Astrophysics Data System (ADS)

Richer, R. A.

2006-12-01

Despite the development of metabolic ecology, the role that carbohydrate stores play in regulating phenological events is unknown. Whole tree manipulation experiments in the common southern African tree species, Brachystegia spiciformis offer new insights into the physiological and environmental mechanisms controlling bud break. This experiment tested a novel hypothesis that decreasing Total Non-Structural Carbohydrates (TNC) in the stem could cue bud break in Brachystegia spiciformis. The experimental treatments included fertilization, canopy defoliation, shading and stem heating and were repeated over a two year period in the Kalahari sand savanna system of northwestern Zimbabwe. The treatments were designed to decrease stem carbohydrates and result in an earlier leaf flush. None of the treatments significantly decreased seasonal stem TNC. Likewise the heating, fertilization and defoliation treatments did not significantly affect the date of bud break from controls. However, shaded trees showed a significant delay in bud break. This delay in bud break could not be attributed to leaf level photosynthetic traits, stem water content, leaf pre-dawn water potential or delayed leaf fall. These results question widely accepted hypotheses about the mechanism controlling bud break in savanna ecosystems and may suggest a carbohydrate homeostatic mechanism.

Study on the change law of hyperspectral data and pigments for rice in mature process

NASA Astrophysics Data System (ADS)

Huang, Jingfeng; Tang, Yanlin; Wang, Renchao

2004-01-01

The hyperspectral reflectances of the canopy, the sword leaf, the third unfolding leaf from the top and ear of the main stem of two varieties of rice are measured by a ASD FieldSpec Pro FR in field and indoor under 3 nitrogen support levels in mature process. The concentrations of chlorophyll and carotenoid of leaves and ears corresponding to the spectra were determined by biochemical method. The spectral differences are significant for the canopy and leaves of rice under differet nitrogen support level, and the concentrations of chlorophyll and carotenoid of leaves increase with the increasing of nitrogen applying. There exist significant differences for the pigment concentrations of the leaves of rice under different nitrogen levels. The spectral reflectances of the canopy are gradually getting bigger in the visible region and smaller in the near infrared region as the growth stage goes on. 'Blue shift' phenomena for the spectra red edge position of the canopy, leaves and ears were proved. The concentrations of chlorophyll and carotenoid of leaves and ears are very significantly correlative to the spectral vegetation indices VI1(= R990/R553), VI2(=R1200/R553), VI3(=R750/R553), VI4(=R670/R440), VI5(= R553/R670), PRVI(=R800/R553), PSSRa, PSNDa and λred (the red edge position). The results show that these VIs can be used to estimate the concentrations of chlorophyll and carotenoid of leaves and ears of rice.

Multiple phenological responses to climate change among 42 plant species in Xi'an, China.

PubMed

Dai, Junhu; Wang, Huanjiong; Ge, Quansheng

2013-09-01

Phenological data of 42 woody plants in a temperate deciduous forest from the Chinese Phenological Observation Network (CPON) and the corresponding meteorological data from 1963 to 2011 in Xi'an, Shaanxi Province, China were collected and analyzed. The first leaf date (FLD), leaf coloring date (LCD) and first flower date (FFD) are revealed as strong biological signals of climatic change. The FLD, LCD and FFD of most species are sensitive to average temperature during a certain period before phenophase onset. Regional precipitation also has a significant impact on phenophases of about half of the species investigated. Affected by climate change, the FLD and FFD of these species have advanced by 5.54 days and 10.20 days on average during 2003-2011 compared with the period 1963-1996, respectively. Meanwhile, the LCD has delayed by 10.59 days, and growing season length has extended 16.13 days. Diverse responses of phenology commonly exist among different species and functional groups during the study period. Especially for FFD, the deviations between the above two periods ranged from -20.68 to -2.79 days; biotic pollination species showed a significantly greater advance than abiotic pollination species. These results were conducive to the understanding of possible changes in both the structure of plant communities and interspecific relationships in the context of climate change.

Cytogenetic analysis and mapping of leaf rust resistance in Aegilops speltoides Tausch derived bread wheat line Selection2427 carrying putative gametocidal gene(s).

PubMed

Niranjana, M; Vinod; Sharma, J B; Mallick, Niharika; Tomar, S M S; Jha, S K

2017-12-01

Leaf rust (Puccinia triticina) is a major biotic stress affecting wheat yields worldwide. Host-plant resistance is the best method for controlling leaf rust. Aegilops speltoides is a good source of resistance against wheat rusts. To date, five Lr genes, Lr28, Lr35, Lr36, Lr47, and Lr51, have been transferred from Ae. speltoides to bread wheat. In Selection2427, a bread wheat introgresed line with Ae. speltoides as the donor parent, a dominant gene for leaf rust resistance was mapped to the long arm of chromosome 3B (LrS2427). None of the Lr genes introgressed from Ae. speltoides have been mapped to chromosome 3B. Since none of the designated seedling leaf rust resistance genes have been located on chromosome 3B, LrS2427 seems to be a novel gene. Selection2427 showed a unique property typical of gametocidal genes, that when crossed to other bread wheat cultivars, the F 1 showed partial pollen sterility and poor seed setting, whilst Selection2427 showed reasonable male and female fertility. Accidental co-transfer of gametocidal genes with LrS2427 may have occurred in Selection2427. Though LrS2427 did not show any segregation distortion and assorted independently of putative gametocidal gene(s), its utilization will be difficult due to the selfish behavior of gametocidal genes.

How Kinetics within the Unfolded State Affects Protein Folding: an Analysis Based on Markov State Models and an Ultra-Long MD Trajectory

PubMed Central

Deng, Nan-jie; Dai, Wei

2013-01-01

Understanding how kinetics in the unfolded state affects protein folding is a fundamentally important yet less well-understood issue. Here we employ three different models to analyze the unfolded landscape and folding kinetics of the miniprotein Trp-cage. The first is a 208 μs explicit solvent molecular dynamics (MD) simulation from D. E. Shaw Research containing tens of folding events. The second is a Markov state model (MSM-MD) constructed from the same ultra-long MD simulation; MSM-MD can be used to generate thousands of folding events. The third is a Markov state model built from temperature replica exchange MD simulations in implicit solvent (MSM-REMD). All the models exhibit multiple folding pathways, and there is a good correspondence between the folding pathways from direct MD and those computed from the MSMs. The unfolded populations interconvert rapidly between extended and collapsed conformations on time scales ≤ 40 ns, compared with the folding time of ≈ 5 μs. The folding rates are independent of where the folding is initiated from within the unfolded ensemble. About 90 % of the unfolded states are sampled within the first 40 μs of the ultra-long MD trajectory, which on average explores ~27 % of the unfolded state ensemble between consecutive folding events. We clustered the folding pathways according to structural similarity into “tubes”, and kinetically partitioned the unfolded state into populations that fold along different tubes. From our analysis of the simulations and a simple kinetic model, we find that when the mixing within the unfolded state is comparable to or faster than folding, the folding waiting times for all the folding tubes are similar and the folding kinetics is essentially single exponential despite the presence of heterogeneous folding paths with non-uniform barriers. When the mixing is much slower than folding, different unfolded populations fold independently leading to non-exponential kinetics. A kinetic partition of the Trp-cage unfolded state is constructed which reveals that different unfolded populations have almost the same probability to fold along any of the multiple folding paths. We are investigating whether the results for the kinetics in the unfolded state of the twenty-residue Trp-cage is representative of larger single domain proteins. PMID:23705683

An Application of Unfolding and Cumulative Item Response Theory Models for Noncognitive Scaling: Examining the Assumptions and Applicability of the Generalized Graded Unfolding Model

ERIC Educational Resources Information Center

Sgammato, Adrienne N.

2009-01-01

This study examined the applicability of a relatively new unidimensional, unfolding item response theory (IRT) model called the generalized graded unfolding model (GGUM; Roberts, Donoghue, & Laughlin, 2000). A total of four scaling methods were applied. Two commonly used cumulative IRT models for polytomous data, the Partial Credit Model and…

Rapid atmospheric transport and large-scale deposition of recently synthesized plant waxes

NASA Astrophysics Data System (ADS)

Nelson, Daniel B.; Ladd, S. Nemiah; Schubert, Carsten J.; Kahmen, Ansgar

2018-02-01

Sedimentary plant wax 2H/1H ratios are important tools for understanding hydroclimate and environmental changes, but large spatial and temporal uncertainties exist about transport mechanisms from ecosystem to sediments. To assess atmospheric pathways, we collected aerosol samples for two years at four locations within a ∼60 km radius in northern Switzerland. We measured n-alkane distributions and 2H/1H ratios in these samples, and from local plants, leaf litter, and soil, as well as surface sediment from six nearby lakes. Increased concentrations and 2H depletion of long odd chain n-alkanes in early summer aerosols indicate that most wax aerosol production occurred shortly after leaf unfolding, when plants synthesize waxes in large quantities. During autumn and winter, aerosols were characterized by degraded n-alkanes lacking chain length preferences diagnostic of recent biosynthesis, and 2H/1H values that were in some cases more than 100‰ higher than growing season values. Despite these seasonal shifts, modeled deposition-weighted average 2H/1H values of long odd chain n-alkanes primarily reflected summer values. This was corroborated by n-alkane 2H/1H values in lake sediments, which were similar to deposition-weighted aerosol values at five of six sites. Atmospheric deposition rates for plant n-alkanes on land were ∼20% of accumulation rates in lakes, suggesting a role for direct deposition to lakes or coastal oceans near similar production sources, and likely a larger role for deposition on land and transport in river systems. This mechanism allows mobilization and transport of large quantities of recently produced waxes as fine-grained material to low energy sedimentation sites over short timescales, even in areas with limited topography. Widespread atmospheric transfer well before leaf senescence also highlights the importance of the isotopic composition of early season source water used to synthesize waxes for the geologic record.

Verification of unfold error estimates in the unfold operator code

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

Fehl, D.L.; Biggs, F.

Spectral unfolding is an inverse mathematical operation that attempts to obtain spectral source information from a set of response functions and data measurements. Several unfold algorithms have appeared over the past 30 years; among them is the unfold operator (UFO) code written at Sandia National Laboratories. In addition to an unfolded spectrum, the UFO code also estimates the unfold uncertainty (error) induced by estimated random uncertainties in the data. In UFO the unfold uncertainty is obtained from the error matrix. This built-in estimate has now been compared to error estimates obtained by running the code in a Monte Carlo fashionmore » with prescribed data distributions (Gaussian deviates). In the test problem studied, data were simulated from an arbitrarily chosen blackbody spectrum (10 keV) and a set of overlapping response functions. The data were assumed to have an imprecision of 5{percent} (standard deviation). One hundred random data sets were generated. The built-in estimate of unfold uncertainty agreed with the Monte Carlo estimate to within the statistical resolution of this relatively small sample size (95{percent} confidence level). A possible 10{percent} bias between the two methods was unresolved. The Monte Carlo technique is also useful in underdetermined problems, for which the error matrix method does not apply. UFO has been applied to the diagnosis of low energy x rays emitted by Z-pinch and ion-beam driven hohlraums. {copyright} {ital 1997 American Institute of Physics.}« less

Inter- and intraspecific variation in leaf economic traits in wheat and maize

PubMed Central

Hale, Christine E; Cerabolini, Bruno E L; Cornelissen, Johannes H C; Craine, Joseph; Gough, William A; Kattge, Jens; Tirona, Cairan K F

2018-01-01

Abstract Leaf Economics Spectrum (LES) trait variation underpins multiple agroecological processes and many prominent crop yield models. While there are numerous independent studies assessing trait variation in crops, to date there have been no comprehensive assessments of intraspecific trait variation (ITV) in LES traits for wheat and maize: the world’s most widespread crops. Using trait databases and peer-reviewed literature, we compiled over 700 records of specific leaf area (SLA), maximum photosynthetic rates (Amax) and leaf nitrogen (N) concentrations, for wheat and maize. We evaluated intraspecific LES trait variation, and intraspecific trait–environment relationships. While wheat and maize occupy the upper 90th percentile of LES trait values observed across a global species pool, ITV ranged widely across the LES in wheat and maize. Fertilization treatments had strong impacts on leaf N, while plant developmental stage (here standardized as the number of days since planting) had strong impacts on Amax; days since planting, N fertilization and irrigation all influenced SLA. When controlling for these factors, intraspecific responses to temperature and precipitation explained 39.4 and 43.7 % of the variation in Amax and SLA, respectively, but only 5.4 % of the variation in leaf N. Despite a long history of domestication in these species, ITV in wheat and maize among and within cultivars remains large. Intraspecific trait variation is a critical consideration to refine regional to global models of agroecosystem structure, function and food security. Considerable opportunities and benefits exist for consolidating a crop trait database for a wider range of domesticated plant species. PMID:29484152

Inter- and intraspecific variation in leaf economic traits in wheat and maize.

PubMed

Martin, Adam R; Hale, Christine E; Cerabolini, Bruno E L; Cornelissen, Johannes H C; Craine, Joseph; Gough, William A; Kattge, Jens; Tirona, Cairan K F

2018-02-01

Leaf Economics Spectrum (LES) trait variation underpins multiple agroecological processes and many prominent crop yield models. While there are numerous independent studies assessing trait variation in crops, to date there have been no comprehensive assessments of intraspecific trait variation (ITV) in LES traits for wheat and maize: the world's most widespread crops. Using trait databases and peer-reviewed literature, we compiled over 700 records of specific leaf area (SLA), maximum photosynthetic rates ( A max ) and leaf nitrogen (N) concentrations, for wheat and maize. We evaluated intraspecific LES trait variation, and intraspecific trait-environment relationships. While wheat and maize occupy the upper 90th percentile of LES trait values observed across a global species pool, ITV ranged widely across the LES in wheat and maize. Fertilization treatments had strong impacts on leaf N, while plant developmental stage (here standardized as the number of days since planting) had strong impacts on A max ; days since planting, N fertilization and irrigation all influenced SLA. When controlling for these factors, intraspecific responses to temperature and precipitation explained 39.4 and 43.7 % of the variation in A max and SLA, respectively, but only 5.4 % of the variation in leaf N. Despite a long history of domestication in these species, ITV in wheat and maize among and within cultivars remains large. Intraspecific trait variation is a critical consideration to refine regional to global models of agroecosystem structure, function and food security. Considerable opportunities and benefits exist for consolidating a crop trait database for a wider range of domesticated plant species.

Titin domains progressively unfolded by force are homogenously distributed along the molecule.

PubMed

Bianco, Pasquale; Mártonfalvi, Zsolt; Naftz, Katalin; Kőszegi, Dorina; Kellermayer, Miklós

2015-07-21

Titin is a giant filamentous protein of the muscle sarcomere in which stretch induces the unfolding of its globular domains. However, the mechanisms of how domains are progressively selected for unfolding and which domains eventually unfold have for long been elusive. Based on force-clamp optical tweezers experiments we report here that, in a paradoxical violation of mechanically driven activation kinetics, neither the global domain unfolding rate, nor the folded-state lifetime distributions of full-length titin are sensitive to force. This paradox is reconciled by a gradient of mechanical stability so that domains are gradually selected for unfolding as the magnitude of the force field increases. Atomic force microscopic screening of extended titin molecules revealed that the unfolded domains are distributed homogenously along the entire length of titin, and this homogeneity is maintained with increasing overstretch. Although the unfolding of domains with progressively increasing mechanical stability makes titin a variable viscosity damper, the spatially randomized variation of domain stability ensures that the induced structural changes are not localized but are distributed along the molecule's length. Titin may thereby provide complex safety mechanims for protecting the sarcomere against structural disintegration under excessive mechanical conditions. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Protein unfolding with a steric trap.

PubMed

Blois, Tracy M; Hong, Heedeok; Kim, Tae H; Bowie, James U

2009-10-07

The study of protein folding requires a method to drive unfolding, which is typically accomplished by altering solution conditions to favor the denatured state. This has the undesirable consequence that the molecular forces responsible for configuring the polypeptide chain are also changed. It would therefore be useful to develop methods that can drive unfolding without the need for destabilizing solvent conditions. Here we introduce a new method to accomplish this goal, which we call steric trapping. In the steric trap method, the target protein is labeled with two biotin tags placed close in space so that both biotin tags can only be bound by streptavidin when the protein unfolds. Thus, binding of the second streptavidin is energetically coupled to unfolding of the target protein. Testing the method on a model protein, dihydrofolate reductase (DHFR), we find that streptavidin binding can drive unfolding and that the apparent binding affinity reports on changes in DHFR stability. Finally, by employing the slow off-rate of wild-type streptavidin, we find that DHFR can be locked in the unfolded state. The steric trap method provides a simple method for studying aspects of protein folding and stability in native solvent conditions, could be used to specifically unfold selected domains, and could be applicable to membrane proteins.

The Proteasomal ATPases Use a Slow but Highly Processive Strategy to Unfold Proteins

PubMed Central

Snoberger, Aaron; Anderson, Raymond T.; Smith, David M.

2017-01-01

All domains of life have ATP-dependent compartmentalized proteases that sequester their peptidase sites on their interior. ATPase complexes will often associate with these compartmentalized proteases in order to unfold and inject substrates into the protease for degradation. Significant effort has been put into understanding how ATP hydrolysis is used to apply force to proteins and cause them to unfold. The unfolding kinetics of the bacterial ATPase, ClpX, have been shown to resemble a fast motor that traps unfolded intermediates as a strategy to unfold proteins. In the present study, we sought to determine if the proteasomal ATPases from eukaryotes and archaea exhibit similar unfolding kinetics. We found that the proteasomal ATPases appear to use a different kinetic strategy for protein unfolding, behaving as a slower but more processive and efficient translocation motor, particularly when encountering a folded domain. We expect that these dissimilarities are due to differences in the ATP binding/exchange cycle, the presence of a trans-arginine finger, or the presence of a threading ring (i.e., the OB domain), which may be used as a rigid platform to pull folded domains against. We speculate that these differences may have evolved due to the differing client pools these machines are expected to encounter. PMID:28421184

Histopathological assessment of the infection of maize leaves by Fusarium graminearum, F. proliferatum, and F. verticillioides.

PubMed

Nguyen, Thi Thanh Xuan; Dehne, Heinz-Wilhelm; Steiner, Ulrike

2016-09-01

Young maize plants were inoculated on unfolded mature leaves and on folded immature leaves with Fusarium graminearum, Fusarium proliferatum, and Fusarium verticillioides suspensions. Infection and symptom development of disease on these asymptomatic mature leaves and immature leaves were then documented. Subcuticular infection was found by the three Fusarium species on both symptomatic and symptomless leaves. The three Fusarium species penetrated the stomata of immature leaves by the formation of appressoria-like structures, infection cushions or by direct penetration. Infection by the three species of Fusarium via stomata is reported here for the first time. The superficial hyphae and re-emerging hyphae of the three species produced conidia. The macroconidia of F. graminearum produced secondary macroconidia and F. proliferatum formed microconidia inside the leaf tissues that sporulated through stomata and trichomes. The infection of maize leaves by the three species of Fusarium and their sporulation may contribute inoculum to cob and kernel infection. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Partitioning of 13C-photosynthate from Spur Leaves during Fruit Growth of Three Japanese Pear (Pyrus pyrifolia) Cultivars Differing in Maturation Date

PubMed Central

ZHANG, CAIXI; TANABE, KENJI; TAMURA, FUMIO; ITAI, AKIHIRO; WANG, SHIPING

2005-01-01

• Background and Aims In fruit crops, fruit size at harvest is an important aspect of quality. With Japanese pears (Pyrus pyrifolia), later maturing cultivars usually have larger fruits than earlier maturing cultivars. It is considered that the supply of photosynthate during fruit development is a critical determinant of size. To assess the interaction of assimilate supply and early/late maturity of cultivars and its effect on final fruit size, the pattern of carbon assimilate partitioning from spur leaves (source) to fruit and other organs (sinks) during fruit growth was investigated using three genotypes differing in maturation date. • Methods Partitioning of photosynthate from spur leaves during fruit growth was investigated by exposure of spurs to 13CO2 and measurement of the change in 13C abundance in dry matter with time. Leaf number and leaf area per spur, fresh fruit weight, cell number and cell size of the mesocarp were measured and used to model the development of the spur leaf and fruit. • Key Results Compared with the earlier-maturing cultivars ‘Shinsui’ and ‘Kousui’, the larger-fruited, later-maturing cultivar ‘Shinsetsu’ had a greater total leaf area per spur, greater source strength (source weight × source specific activity), with more 13C assimilated per spur and allocated to fruit, smaller loss of 13C in respiration and export over the season, and longer duration of cell division and enlargement. Histology shows that cultivar differences in final fruit size were mainly attributable to the number of cells in the mesocarp. • Conclusions Assimilate availability during the period of cell division was crucial for early fruit growth and closely correlated with final fruit size. Early fruit growth of the earlier-maturing cultivars, but not the later-maturing ones, was severely restrained by assimilate supply rather than by sink limitation. PMID:15655106

Probing the Structure of the Escherichia coli Periplasmic Proteins HdeA and YmgD by Molecular Dynamics Simulations.

PubMed

Socher, Eileen; Sticht, Heinrich

2016-11-23

HdeA and YmgD are structurally homologous proteins in the periplasm of Escherichia coli. HdeA has been shown to represent an acid-activated chaperone, whereas the function of YmgD has not yet been characterized. We performed pH-titrating molecular dynamics simulations (pHtMD) to investigate the structural changes of both proteins and to assess whether YmgD may also exhibit an unfolding behavior similar to that of HdeA. The unfolding pathway of HdeA includes partially unfolded dimer structures, which represent a prerequisite for subsequent dissociation. In contrast to the coupled unfolding and dissociation of HdeA, YmgD displays dissociation of the folded subunits, and the subunits do not undergo significant unfolding even at low pH values. The differences in subunit stability between HdeA and YmgD may be explained by the structural features of helix D, which represents the starting point of unfolding in HdeA. In summary, the present study suggests that YmgD either is not an acid-activated chaperone or, at least, does not require unfolding for activation.

Unfolding of Proteins: Thermal and Mechanical Unfolding

NASA Technical Reports Server (NTRS)

Hur, Joe S.; Darve, Eric

2004-01-01

We have employed a Hamiltonian model based on a self-consistent Gaussian appoximation to examine the unfolding process of proteins in external - both mechanical and thermal - force elds. The motivation was to investigate the unfolding pathways of proteins by including only the essence of the important interactions of the native-state topology. Furthermore, if such a model can indeed correctly predict the physics of protein unfolding, it can complement more computationally expensive simulations and theoretical work. The self-consistent Gaussian approximation by Micheletti et al. has been incorporated in our model to make the model mathematically tractable by signi cantly reducing the computational cost. All thermodynamic properties and pair contact probabilities are calculated by simply evaluating the values of a series of Incomplete Gamma functions in an iterative manner. We have compared our results to previous molecular dynamics simulation and experimental data for the mechanical unfolding of the giant muscle protein Titin (1TIT). Our model, especially in light of its simplicity and excellent agreement with experiment and simulation, demonstrates the basic physical elements necessary to capture the mechanism of protein unfolding in an external force field.

77 FR 16145 - Airworthiness Directives; Bombardier, Inc.

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-20

... chamfer of the upper edge of each leaf spring, and rework if necessary. This AD also requires installing a... [rework] the springs as applicable, and SB 8-76-02 Rev. A to replace the pre-SB 8-76-2 (Mod 8/ 0443... Bulletin A8-76-32, dated January 27, 2012. Do all applicable rework before further flight, in accordance...

Leafing-out date not indicative of growth rate in hybrid poplars

Treesearch

Harold F. Ford; Edward I. Sucoff

1961-01-01

In breeding trees for rapid growth, testing the progeny usually requires many years. To shorten the testing period, geneticists have tried to find characteristics in juvenile trees that would indicate mature-tree performance. With hybrid poplars (Populus spp.), work at the Northeastern Forest Experiment Station has shown that the thickness of bark on...

Unfolding of the bacterial nucleoid both in vivo and in vitro as a result of exposure to camphor.

PubMed Central

Harrington, E W; Trun, N J

1997-01-01

Both prokaryotic and eukaryotic cells are sensitive to killing by camphor; however, the mechanism by which camphor kills has not been elucidated. We report here that camphor unfolds the nucleoid of Escherichia coli and that unfolding does not require DNA replication, translation, or cell division. We show that exposure of isolated nucleoids to camphor results in unfolding of the chromosome. PMID:9079934

Machinery of protein folding and unfolding.

PubMed

Zhang, Xiaodong; Beuron, Fabienne; Freemont, Paul S

2002-04-01

During the past two years, a large amount of biochemical, biophysical and low- to high-resolution structural data have provided mechanistic insights into the machinery of protein folding and unfolding. It has emerged that dual functionality in terms of folding and unfolding might exist for some systems. The majority of folding/unfolding machines adopt oligomeric ring structures in a cooperative fashion and utilise the conformational changes induced by ATP binding/hydrolysis for their specific functions.

Single molecule unfolding and stretching of protein domains inside a solid-state nanopore by electric field.

PubMed

Freedman, Kevin J; Haq, S Raza; Edel, Joshua B; Jemth, Per; Kim, Min Jun

2013-01-01

Single molecule methods have provided a significantly new look at the behavior of biomolecules in both equilibrium and non-equilibrium conditions. Most notable are the stretching experiments performed by atomic force microscopes and laser tweezers. Here we present an alternative single molecule method that can unfold a protein domain, observed at electric fields greater than 10(6) V/m, and is fully controllable by the application of increasing voltages across the membrane of the pore. Furthermore this unfolding mechanism is characterized by measuring both the residence time of the protein within the nanopore and the current blockade. The unfolding data supports a gradual unfolding mechanism rather than the cooperative transition observed by classical urea denaturation experiments. Lastly it is shown that the voltage-mediated unfolding is a function of the stability of the protein by comparing two mutationally destabilized variants of the protein.

UNFOLD-SENSE: a parallel MRI method with self-calibration and artifact suppression.

PubMed

Madore, Bruno

2004-08-01

This work aims at improving the performance of parallel imaging by using it with our "unaliasing by Fourier-encoding the overlaps in the temporal dimension" (UNFOLD) temporal strategy. A self-calibration method called "self, hybrid referencing with UNFOLD and GRAPPA" (SHRUG) is presented. SHRUG combines the UNFOLD-based sensitivity mapping strategy introduced in the TSENSE method by Kellman et al. (5), with the strategy introduced in the GRAPPA method by Griswold et al. (10). SHRUG merges the two approaches to alleviate their respective limitations, and provides fast self-calibration at any given acceleration factor. UNFOLD-SENSE further includes an UNFOLD artifact suppression scheme to significantly suppress artifacts and amplified noise produced by parallel imaging. This suppression scheme, which was published previously (4), is related to another method that was presented independently as part of TSENSE. While the two are equivalent at accelerations < or = 2.0, the present approach is shown here to be significantly superior at accelerations > 2.0, with up to double the artifact suppression at high accelerations. Furthermore, a slight modification of Cartesian SENSE is introduced, which allows departures from purely Cartesian sampling grids. This technique, termed variable-density SENSE (vdSENSE), allows the variable-density data required by SHRUG to be reconstructed with the simplicity and fast processing of Cartesian SENSE. UNFOLD-SENSE is given by the combination of SHRUG for sensitivity mapping, vdSENSE for reconstruction, and UNFOLD for artifact/amplified noise suppression. The method was implemented, with online reconstruction, on both an SSFP and a myocardium-perfusion sequence. The results from six patients scanned with UNFOLD-SENSE are presented.

Salt bridge as a gatekeeper against partial unfolding.

PubMed

Hinzman, Mark W; Essex, Morgan E; Park, Chiwook

2016-05-01

Salt bridges are frequently observed in protein structures. Because the energetic contribution of salt bridges is strongly dependent on the environmental context, salt bridges are believed to contribute to the structural specificity rather than the stability. To test the role of salt bridges in enhancing structural specificity, we investigated the contribution of a salt bridge to the energetics of native-state partial unfolding in a cysteine-free version of Escherichia coli ribonuclease H (RNase H*). Thermolysin cleaves a protruding loop of RNase H(*) through transient partial unfolding under native conditions. Lys86 and Asp108 in RNase H(*) form a partially buried salt bridge that tethers the protruding loop. Investigation of the global stability of K86Q/D108N RNase H(*) showed that the salt bridge does not significantly contribute to the global stability. However, K86Q/D108N RNase H(*) is greatly more susceptible to proteolysis by thermolysin than wild-type RNase H(*) is. The free energy for partial unfolding determined by native-state proteolysis indicates that the salt bridge significantly increases the energy for partial unfolding by destabilizing the partially unfolded form. Double mutant cycles with single and double mutations of the salt bridge suggest that the partially unfolded form is destabilized due to a significant decrease in the interaction energy between Lys86 and Asp108 upon partial unfolding. This study demonstrates that, even in the case that a salt bridge does not contribute to the global stability, the salt bridge may function as a gatekeeper against partial unfolding that disturbs the optimal geometry of the salt bridge. © 2016 The Protein Society.

Protein unfolding versus β-sheet separation in spider silk nanocrystals

NASA Astrophysics Data System (ADS)

Alam, Parvez

2014-03-01

In this communication a mechanism for spider silk strain hardening is proposed. Shear failure of β-sheet nanocrystals is the first failure mode that gives rise to the creation of smaller nanocrystals, which are of higher strength and stiffness. β-sheet unfolding requires more energy than nanocrystal separation in a shear mode of failure. As a result, unfolding occurs after the nanocrystals separate in shear. β-sheet unfolding yields a secondary strain hardening effect once the β-sheet conformation is geometrically stable and acts like a unidirectional fibre in a fibre reinforced composite. The mechanism suggested herein is based on molecular dynamics calculations of residual inter-β-sheet separation strengths against residual intra-β-sheet unfolding strengths.

Comparative Study of the Mechanical Unfolding Pathways of α- and β-Peptides.

PubMed

Uribe, Lalita; Gauss, Jürgen; Diezemann, Gregor

2015-07-02

Using molecular simulations, we analyze the unfolding pathways of various peptides. We compare the mechanical unfolding of a β-alanine's octamer (β-HAla8) and an α-alanine's decamer (α-Ala10). Using force-probe molecular-dynamics simulations, to induce unfolding, we show that the 3(14)-helix formed by β-HAla8 is mechanically more stable than the α-helix formed by α-Ala10, although both structures are stabilized by six hydrogen bonds. Additionally, computations of the potential of mean force validate this result and show that also the thermal stability of the 3(14)-helix is higher. It is demonstrated that β-HAla8 unfolds in a two-step fashion with a stable intermediate. This is contrasted with the known single-step scenario of the unfolding of α-Ala10. Furthermore, we present a study of the chain-length dependence of the mechanical unfolding pathway of the 3(14)-helix. The calculation of the dynamic strength for oligomers with chain lengths ranging from 6 to 18 monomers shows that the unfolding pathway of helices with an integer and noninteger number of turns has m + 1 and m energy barriers, respectively, with m being the number of complete turns. The additional barrier for helices with an integer number of turns is shown to be related to the breaking of the N-terminus' hydrogen bond.

Resolution of the unfolded state.

NASA Astrophysics Data System (ADS)

Beaucage, Gregory

2008-03-01

The unfolded states in proteins and nucleic acids remain weakly understood despite their importance to protein folding; misfolding diseases (Parkinson's & Alzheimer's); natively unfolded proteins (˜ 30% of eukaryotic proteins); and to understanding ribozymes. Research has been hindered by the inability to quantify the residual (native) structure present in an unfolded protein or nucleic acid. Here, a scaling model is proposed to quantify the degree of folding and the unfolded state (Beaucage, 2004, 2007). The model takes a global view of protein structure and can be applied to a number of analytic methods and to simulations. Three examples are given of application to small-angle scattering from pressure induced unfolding of SNase (Panick, 1998), from acid unfolded Cyt c (Kataoka, 1993) and from folding of Azoarcus ribozyme (Perez-Salas, 2004). These examples quantitatively show 3 characteristic unfolded states for proteins, the statistical nature of a folding pathway and the relationship between extent of folding and chain size during folding for charge driven folding in RNA. Beaucage, G., Biophys. J., in press (2007). Beaucage, G., Phys. Rev. E. 70, 031401 (2004). Kataoka, M., Y. Hagihara, K. Mihara, Y. Goto J. Mol. Biol. 229, 591 (1993). Panick, G., R. Malessa, R. Winter, G. Rapp, K. J. Frye, C. A. Royer J. Mol. Biol. 275, 389 (1998). Perez-Salas U. A., P. Rangan, S. Krueger, R. M. Briber, D. Thirumalai, S. A. Woodson, Biochemistry 43 1746 (2004).

TFE-induced local unfolding and fibrillation of SOD1: bridging the experiment and simulation studies.

PubMed

Kumar, Vijay; Prakash, Amresh; Pandey, Preeti; Lynn, Andrew M; Hassan, Md Imtaiyaz

2018-05-18

Misfolding and aggregation of Cu, Zn Superoxide dismutase (SOD1) is involved in the neurodegenerative disease, amyotrophic lateral sclerosis. Many studies have shown that metal-depleted, monomeric form of SOD1 displays substantial local unfolding dynamics and is the precursor for aggregation. Here, we have studied the structure and dynamics of different apo monomeric SOD1 variants associated with unfolding and aggregation in aqueous trifluoroethanol (TFE) through experiments and simulation. TFE induces partially unfolded β-sheet-rich extended conformations in these SOD1 variants, which subsequently develops aggregates with fibril-like characteristics. Fibrillation was achieved more easily in disulfide-reduced monomeric SOD1 when compared with wild-type and mutant monomeric SOD1. At higher concentrations of TFE, a native-like structure with the increase in α-helical content was observed. The molecular dynamics simulation results illustrate distinct structural dynamics for different regions of SOD1 variants and show uniform local unfolding of β-strands. The strands protected by the zinc-binding and electrostatic loops were found to unfold first in 20% (v/v) TFE, leading to a partial unfolding of β-strands 4, 5, and 6 which are prone to aggregation. Our results thus shed light on the role of local unfolding and conformational dynamics in SOD1 misfolding and aggregation. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Mature phenotype in Hemerocallis plantlets fortuitously generated in vitro

NASA Technical Reports Server (NTRS)

Fitter, M. S.; Krikorian, A. D.

1985-01-01

Daylily plantlets generated on semi-solid media from morphogenetically competent cells or morphogenetically competent cells regenerated from protoplasts can give rise in aseptic culture to plantlets with a mature phenotype. The individual leaves of these plantlets open to the extreme base so that no encircling leaf sheath is present. This permits the overlapping bases and leaves to assume an open fan-like arrangement. The occurrence of fans correlates with exceptionally tightly sealed culture vessels and experiments to date suggest a gaseous component is associated with this change of growth form. It has not been possible to fix the mature growth mode, however, and new leaf growth assumes the more normal juvenile phenotype when the gaseous environment is altered by admitting or exposure to room air.

Phenotypic and biochemical profile changes in calendula (Calendula officinalis L.) plants treated with two chemical mutagenesis.

PubMed

El-Nashar, Y I; Asrar, A A

2016-05-06

Chemical mutagenesis is an efficient tool used in mutation-breeding programs to improve the vital characters of the floricultural crops. This study aimed to estimate the effects of different concentrations of two chemical mutagens; sodium azide (SA) and diethyl sulfate (DES). The vegetative growth and flowering characteristics in two generations (M1 and M2) of calendula plants were investigated. Seeds were treated with five different concentrations of SA and DES (at the same rates) of 1000, 2000, 3000, 4000, and 5000 ppm, in addition to a control treatment of 0 ppm. Results showed that lower concentrations of SA mutagen had significant effects on seed germination percentage, plant height, leaf area, plant fresh weight, flowering date, inflorescence diameter, and gas-exchange measurements in plants of both generations. Calendula plants tended to flower earlier under low mutagen concentrations (1000 ppm), whereas higher concentrations delayed flowering significantly. Positive results on seed germination, plant height, number of branches, plant fresh weight, and leaf area were observed in the M2-generation at lower concentrations of SA (1000 ppm), as well as at 4000 ppm DES on number of leaves and inflorescences. The highest total soluble protein was detected at the concentrations of 1000 ppm SA and 2000 ppm DES. DES showed higher average of acid phosphatase activity than SA. Results indicated that lower concentrations of SA and DES mutagens had positive effects on seed germination percentage, plant height, leaf area, plant fresh weight, flowering date, inflorescence diameter, and gas-exchange measurements. Thus, lower mutagen concentrations could be recommended for better floral and physio-chemical performance.

Chromosome sizes of phytoplasmas composing major phylogenetic groups and subgroups.

PubMed

Marcone, C; Neimark, H; Ragozzino, A; Lauer, U; Seemüller, E

1999-09-01

ABSTRACT Chromosome sizes of 71 phytoplasmas belonging to 12 major phylogenetic groups including several of the aster yellows subgroups were estimated from electrophoretic mobilities of full-length chromosomes in pulsed-field gels. Considerable variation in genome size, from 660 to 1,130 kilobases (kb), was observed among aster yellows phytoplasmas. Chromosome size heterogeneity was also observed in the stolbur phytoplasma group (range 860 to 1,350 kb); in this group, isolate STOLF contains the largest chromosome found in a phytoplasma to date. A wide range of chromosome sizes, from 670 to 1,075 kb, was also identified in the X-disease group. The other phytoplasmas examined, which included members of the apple proliferation, Italian alfalfa witches' broom, faba bean phyllody, pigeon pea witches' broom, sugarcane white leaf, Bermuda grass white leaf, ash yellows, clover proliferation, and elm yellows groups, all have chromosomes smaller than 1 megabase, and the size ranges within each of these groups is narrower than in the aster yellows, stolbur, and X-disease groups. The smallest chromosome, approximately 530 kb, was found in two Bermuda grass white leaf phytoplasma isolates. This not only is the smallest mollicute chromosome found to date, but also is the smallest chromosome known for any cell. More than one large DNA band was observed in several phytoplasma preparations. Possible explanations for the occurrence of more than one band may be infection of the host plant by different phytoplasmas, the presence of more than one chromosome in the same organism, or the presence of large extrachromosomal DNA elements.

Probing the folded state and mechanical unfolding pathways of T4 lysozyme using all-atom and coarse-grained molecular simulation

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

Zheng, Wenjun, E-mail: wjzheng@buffalo.edu; Glenn, Paul

2015-01-21

The Bacteriophage T4 Lysozyme (T4L) is a prototype modular protein comprised of an N-terminal and a C-domain domain, which was extensively studied to understand the folding/unfolding mechanism of modular proteins. To offer detailed structural and dynamic insights to the folded-state stability and the mechanical unfolding behaviors of T4L, we have performed extensive equilibrium and steered molecular dynamics simulations of both the wild-type (WT) and a circular permutation (CP) variant of T4L using all-atom and coarse-grained force fields. Our all-atom and coarse-grained simulations of the folded state have consistently found greater stability of the C-domain than the N-domain in isolation, whichmore » is in agreement with past thermostatic studies of T4L. While the all-atom simulation cannot fully explain the mechanical unfolding behaviors of the WT and the CP variant observed in an optical tweezers study, the coarse-grained simulations based on the Go model or a modified elastic network model (mENM) are in qualitative agreement with the experimental finding of greater unfolding cooperativity in the WT than the CP variant. Interestingly, the two coarse-grained models predict different structural mechanisms for the observed change in cooperativity between the WT and the CP variant—while the Go model predicts minor modification of the unfolding pathways by circular permutation (i.e., preserving the general order that the N-domain unfolds before the C-domain), the mENM predicts a dramatic change in unfolding pathways (e.g., different order of N/C-domain unfolding in the WT and the CP variant). Based on our simulations, we have analyzed the limitations of and the key differences between these models and offered testable predictions for future experiments to resolve the structural mechanism for cooperative folding/unfolding of T4L.« less

A new atomic force microscope force ramp technique using digital force feedback control reveals mechanically weak protein unfolding events.

PubMed

Kawakami, M; Smith, D A

2008-12-10

We have developed a new force ramp modification of the atomic force microscope (AFM) which can control multiple unfolding events of a multi-modular protein using software-based digital force feedback control. With this feedback the force loading rate can be kept constant regardless the length of soft elastic linkage or number of unfolded polypeptide domains. An unfolding event is detected as a sudden drop in force, immediately after which the feedback control reduces the applied force to a low value of a few pN by lowering the force set point. Hence the remaining folded domains can relax and the subsequent force ramp is applied to relaxed protein domains identically in each case. We have applied this technique to determine the kinetic parameters x(u), which is the distance between the native state and transition state, and α(0), which is the unfolding rate constant at zero force, for the mechanical unfolding of a pentamer of I27 domains of titin. In each force ramp the unfolding probability depends on the number of folded domains remaining in the system and we had to take account of this effect in the analysis of unfolding force data. We obtained values of x(u) and α(0) to be 0.28 nm and 1.02 × 10(-3) s(-1), which are in good agreement with those obtained from conventional constant velocity experiments. This method reveals unfolding data at low forces that are not seen in constant velocity experiments and corrects for the change in stiffness that occurs with most mechanical systems throughout the unfolding process to allow constant force ramp experiments to be carried out. In addition, a mechanically weak structure was detected, which formed from the fully extended polypeptide chain during a force quench. This indicates that the new technique will allow studies of the folding kinetics of previously hidden, mechanically weak species.

OPERATOR NORM INEQUALITIES BETWEEN TENSOR UNFOLDINGS ON THE PARTITION LATTICE

PubMed Central

Wang, Miaoyan; Duc, Khanh Dao; Fischer, Jonathan; Song, Yun S.

2017-01-01

Interest in higher-order tensors has recently surged in data-intensive fields, with a wide range of applications including image processing, blind source separation, community detection, and feature extraction. A common paradigm in tensor-related algorithms advocates unfolding (or flattening) the tensor into a matrix and applying classical methods developed for matrices. Despite the popularity of such techniques, how the functional properties of a tensor changes upon unfolding is currently not well understood. In contrast to the body of existing work which has focused almost exclusively on matricizations, we here consider all possible unfoldings of an order-k tensor, which are in one-to-one correspondence with the set of partitions of {1, …, k}. We derive general inequalities between the lp-norms of arbitrary unfoldings defined on the partition lattice. In particular, we demonstrate how the spectral norm (p = 2) of a tensor is bounded by that of its unfoldings, and obtain an improved upper bound on the ratio of the Frobenius norm to the spectral norm of an arbitrary tensor. For specially-structured tensors satisfying a generalized definition of orthogonal decomposability, we prove that the spectral norm remains invariant under specific subsets of unfolding operations. PMID:28286347

(Un)Folding Mechanisms of the FBP28 WW Domain in Explicit Solvent Revealed by Multiple Rare Event Simulation Methods

PubMed Central

Juraszek, Jarek; Bolhuis, Peter G.

2010-01-01

Abstract We report a numerical study of the (un)folding routes of the truncated FBP28 WW domain at ambient conditions using a combination of four advanced rare event molecular simulation techniques. We explore the free energy landscape of the native state, the unfolded state, and possible intermediates, with replica exchange molecular dynamics. Subsequent application of bias-exchange metadynamics yields three tentative unfolding pathways at room temperature. Using these paths to initiate a transition path sampling simulation reveals the existence of two major folding routes, differing in the formation order of the two main hairpins, and in hydrophobic side-chain interactions. Having established that the hairpin strand separation distances can act as reasonable reaction coordinates, we employ metadynamics to compute the unfolding barriers and find that the barrier with the lowest free energy corresponds with the most likely pathway found by transition path sampling. The unfolding barrier at 300 K is ∼17 kBT ≈ 42 kJ/mol, in agreement with the experimental unfolding rate constant. This work shows that combining several powerful simulation techniques provides a more complete understanding of the kinetic mechanism of protein folding. PMID:20159161

Verification of unfold error estimates in the UFO code

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

Fehl, D.L.; Biggs, F.

Spectral unfolding is an inverse mathematical operation which attempts to obtain spectral source information from a set of tabulated response functions and data measurements. Several unfold algorithms have appeared over the past 30 years; among them is the UFO (UnFold Operator) code. In addition to an unfolded spectrum, UFO also estimates the unfold uncertainty (error) induced by running the code in a Monte Carlo fashion with prescribed data distributions (Gaussian deviates). In the problem studied, data were simulated from an arbitrarily chosen blackbody spectrum (10 keV) and a set of overlapping response functions. The data were assumed to have anmore » imprecision of 5% (standard deviation). 100 random data sets were generated. The built-in estimate of unfold uncertainty agreed with the Monte Carlo estimate to within the statistical resolution of this relatively small sample size (95% confidence level). A possible 10% bias between the two methods was unresolved. The Monte Carlo technique is also useful in underdetemined problems, for which the error matrix method does not apply. UFO has been applied to the diagnosis of low energy x rays emitted by Z-Pinch and ion-beam driven hohlraums.« less

Cooperative unfolding of distinctive mechanoreceptor domains transduces force into signals

PubMed Central

Ju, Lining; Chen, Yunfeng; Xue, Lingzhou; Du, Xiaoping; Zhu, Cheng

2016-01-01

How cells sense their mechanical environment and transduce forces into biochemical signals is a crucial yet unresolved question in mechanobiology. Platelets use receptor glycoprotein Ib (GPIb), specifically its α subunit (GPIbα), to signal as they tether and translocate on von Willebrand factor (VWF) of injured arterial surfaces against blood flow. Force elicits catch bonds to slow VWF–GPIbα dissociation and unfolds the GPIbα leucine-rich repeat domain (LRRD) and juxtamembrane mechanosensitive domain (MSD). How these mechanical processes trigger biochemical signals remains unknown. Here we analyze these extracellular events and the resulting intracellular Ca2+ on a single platelet in real time, revealing that LRRD unfolding intensifies Ca2+ signal whereas MSD unfolding affects the type of Ca2+ signal. Therefore, LRRD and MSD are analog and digital force transducers, respectively. The >30 nm macroglycopeptide separating the two domains transmits force on the VWF–GPIbα bond (whose lifetime is prolonged by LRRD unfolding) to the MSD to enhance its unfolding, resulting in unfolding cooperativity at an optimal force. These elements may provide design principles for a generic mechanosensory protein machine. DOI: http://dx.doi.org/10.7554/eLife.15447.001 PMID:27434669

OPERATOR NORM INEQUALITIES BETWEEN TENSOR UNFOLDINGS ON THE PARTITION LATTICE.

PubMed

Wang, Miaoyan; Duc, Khanh Dao; Fischer, Jonathan; Song, Yun S

2017-05-01

Interest in higher-order tensors has recently surged in data-intensive fields, with a wide range of applications including image processing, blind source separation, community detection, and feature extraction. A common paradigm in tensor-related algorithms advocates unfolding (or flattening) the tensor into a matrix and applying classical methods developed for matrices. Despite the popularity of such techniques, how the functional properties of a tensor changes upon unfolding is currently not well understood. In contrast to the body of existing work which has focused almost exclusively on matricizations, we here consider all possible unfoldings of an order- k tensor, which are in one-to-one correspondence with the set of partitions of {1, …, k }. We derive general inequalities between the l p -norms of arbitrary unfoldings defined on the partition lattice. In particular, we demonstrate how the spectral norm ( p = 2) of a tensor is bounded by that of its unfoldings, and obtain an improved upper bound on the ratio of the Frobenius norm to the spectral norm of an arbitrary tensor. For specially-structured tensors satisfying a generalized definition of orthogonal decomposability, we prove that the spectral norm remains invariant under specific subsets of unfolding operations.

Thermodynamics of the Trp-cage Miniprotein Unfolding in Urea

PubMed Central

Wafer, Lucas N. R.; Streicher, Werner W.; Makhatadze, George I.

2010-01-01

The thermodynamic properties of unfolding of the Trp-cage mini protein in the presence of various concentrations of urea have been characterized using temperature-induced unfolding monitored by far-UV circular dichroism spectroscopy. Analysis of the data using a two-state model allowed the calculation of the Gibbs energy of unfolding at 25°C as a function of urea concentration. This in turn was analyzed by the linear extrapolation model that yielded the dependence of Gibbs energy on urea concentration, i.e. the m-value for Trp-cage unfolding. The m-value obtained from the experimental data, as well as the experimental heat capacity change upon unfolding, were correlated with the structural parameters derived from the three dimensional structure of Trp-cage. It is shown that the m-value can be predicted well using a transfer model, while the heat capacity changes are in very good agreement with the empirical models based on model compounds studies. These results provide direct evidence that Trp-cage, despite its small size, is an excellent model for studies of protein unfolding and provide thermodynamic data that can be used to compare with atomistic computer simulations. PMID:20112418

Functional Inactivation of Putative Photosynthetic Electron Acceptor Ferredoxin C2 (FdC2) Induces Delayed Heading Date and Decreased Photosynthetic Rate in Rice

PubMed Central

Ruan, Banpu; Kang, Shujing; He, Lei; Zhang, Sen; Dong, Guojun; Hu, Jiang; Zeng, Dali; Zhang, Guangheng; Gao, Zhenyu; Ren, Deyong; Hu, Xingming; Chen, Guang; Guo, Longbiao; Qian, Qian; Zhu, Li

2015-01-01

Ferredoxin (Fd) protein as unique electron acceptor, involved in a variety of fundamental metabolic and signaling processes, which is indispensable for plant growth. The molecular mechanisms of Fd such as regulation of electron partitioning, impact of photosynthetic rate and involvement in the carbon fixing remain elusive in rice. Here we reported a heading date delay and yellowish leaf 1 (hdy1) mutant derived from Japonica rice cultivar “Nipponbare” subjected to EMS treatment. In the paddy field, the hdy1 mutant appeared at a significantly late heading date and had yellow-green leaves during the whole growth stage. Further investigation indicated that the abnormal phenotype of hdy1 was connected with depressed pigment content and photosynthetic rate. Genetic analysis results showed that the hdy1 mutant phenotype was caused by a single recessive nuclear gene mutation. Map-based cloning revealed that OsHDY1 is located on chromosome 3 and encodes an ortholog of the AtFdC2 gene. Complementation and overexpression, transgenic plants exhibited the mutant phenotype including head date, leaf color and the transcription levels of the FdC2 were completely rescued by transformation with OsHDY1. Real-time PCR revealed that the expression product of OsHDY1 was detected in almost all of the organs except root, whereas highest expression levels were observed in seeding new leaves. The lower expression levels of HDY1 and content of iron were detected in hdy1 than WT’s. The FdC2::GFP was detected in the chloroplasts of rice. Real-time PCR results showed that the expression of many photosynthetic electron transfer related genes in hdy1 were higher than WT. Our results suggest that OsFdC2 plays an important role in photosynthetic rate and development of heading date by regulating electron transfer and chlorophyll content in rice. PMID:26598971

Stereochemistry and solvent role in protein folding: nuclear magnetic resonance and molecular dynamics studies of poly-L and alternating-L,D homopolypeptides in dimethyl sulfoxide.

PubMed

Srivastava, Kinshuk Raj; Kumar, Anil; Goyal, Bhupesh; Durani, Susheel

2011-05-26

The competing interactions folding and unfolding protein structure remain obscure. Using homopolypeptides, we ask if poly-L structure may have a role. We mutate the structure to alternating-L,D stereochemistry and substitute water as the fold-promoting solvent with methanol and dimethyl sulfoxide (DMSO) as the fold-denaturing solvents. Circular dichroism and molecular dynamics established previously that, while both isomers were folded in water, the poly-L isomer was unfolded and alternating-L,D isomer folded in methanol. Nuclear magnetic resonance and molecular dynamics establish now that both isomers are unfolded in DMSO. We calculated energetics of folding-unfolding equilibrium with water and methanol as solvents. We have now calculated interactions of unfolded polypeptide structures with DMSO as solvent. Methanol was found to unfold and water fold poly-L structure as a dielectric. DMSO has now been found to unfold both poly-L and alternating-L,D structures by strong solvation of peptides to disrupt their hydrogen bonds. Accordingly, we propose that while linked peptides fold protein structure with hydrogen bonds they unfold the structure electrostatically due to the stereochemical effect of the poly-L structure. Protein folding to ordering of peptide hydrogen bonds with water as canonical solvent may thus involve two specific and independent solvent effects-one, strong screening of electrostatics of poly-L linked peptides, and two, weak dipolar solvation of peptides. Correspondingly, protein denaturation may involve two independent solvent effects-one, weak dielectric to unfold poly-L structure electrostatically, and two, strong polarity to disrupt peptide hydrogen bonds by solvation of peptides.

As Simple As Possible, but Not Simpler: Exploring the Fidelity of Coarse-Grained Protein Models for Simulated Force Spectroscopy

PubMed Central

Rottler, Jörg; Plotkin, Steven S.

2016-01-01

Mechanical unfolding of a single domain of loop-truncated superoxide dismutase protein has been simulated via force spectroscopy techniques with both all-atom (AA) models and several coarse-grained models having different levels of resolution: A Gō model containing all heavy atoms in the protein (HA-Gō), the associative memory, water mediated, structure and energy model (AWSEM) which has 3 interaction sites per amino acid, and a Gō model containing only one interaction site per amino acid at the Cα position (Cα-Gō). To systematically compare results across models, the scales of time, energy, and force had to be suitably renormalized in each model. Surprisingly, the HA-Gō model gives the softest protein, exhibiting much smaller force peaks than all other models after the above renormalization. Clustering to render a structural taxonomy as the protein unfolds showed that the AA, HA-Gō, and Cα-Gō models exhibit a single pathway for early unfolding, which eventually bifurcates repeatedly to multiple branches only after the protein is about half-unfolded. The AWSEM model shows a single dominant unfolding pathway over the whole range of unfolding, in contrast to all other models. TM alignment, clustering analysis, and native contact maps show that the AWSEM pathway has however the most structural similarity to the AA model at high nativeness, but the least structural similarity to the AA model at low nativeness. In comparison to the AA model, the sequence of native contact breakage is best predicted by the HA-Gō model. All models consistently predict a similar unfolding mechanism for early force-induced unfolding events, but diverge in their predictions for late stage unfolding events when the protein is more significantly disordered. PMID:27898663

Unfolding of Ubiquitin Studied by Picosecond Time-Resolved Fluorescence of the Tyrosine Residue

PubMed Central

Noronha, Melinda; Lima, João C.; Bastos, Margarida; Santos, Helena; Maçanita, António L.

2004-01-01

The photophysics of the single tyrosine in bovine ubiquitin (UBQ) was studied by picosecond time-resolved fluorescence spectroscopy, as a function of pH and along thermal and chemical unfolding, with the following results: First, at room temperature (25°C) and below pH 1.5, native UBQ shows single-exponential decays. From pH 2 to 7, triple-exponential decays were observed and the three decay times were attributed to the presence of tyrosine, a tyrosine-carboxylate hydrogen-bonded complex, and excited-state tyrosinate. Second, at pH 1.5, the water-exposed tyrosine of either thermally or chemically unfolded UBQ decays as a sum of two exponentials. The double-exponential decays were interpreted and analyzed in terms of excited-state intramolecular electron transfer from the phenol to the amide moiety, occurring in one of the three rotamers of tyrosine in UBQ. The values of the rate constants indicate the presence of different unfolded states and an increase in the mobility of the tyrosine residue during unfolding. Finally, from the pre-exponential coefficients of the fluorescence decays, the unfolding equilibrium constants (KU) were calculated, as a function of temperature or denaturant concentration. Despite the presence of different unfolded states, both thermal and chemical unfolding data of UBQ could be fitted to a two-state model. The thermodynamic parameters Tm = 54.6°C, ΔHTm = 56.5 kcal/mol, and ΔCp = 890 cal/mol//K, were determined from the unfolding equilibrium constants calculated accordingly, and compared to values obtained by differential scanning calorimetry also under the assumption of a two-state transition, Tm = 57.0°C, ΔHm= 51.4 kcal/mol, and ΔCp = 730 cal/mol//K. PMID:15454455

As Simple As Possible, but Not Simpler: Exploring the Fidelity of Coarse-Grained Protein Models for Simulated Force Spectroscopy.

PubMed

Habibi, Mona; Rottler, Jörg; Plotkin, Steven S

2016-11-01

Mechanical unfolding of a single domain of loop-truncated superoxide dismutase protein has been simulated via force spectroscopy techniques with both all-atom (AA) models and several coarse-grained models having different levels of resolution: A Gō model containing all heavy atoms in the protein (HA-Gō), the associative memory, water mediated, structure and energy model (AWSEM) which has 3 interaction sites per amino acid, and a Gō model containing only one interaction site per amino acid at the Cα position (Cα-Gō). To systematically compare results across models, the scales of time, energy, and force had to be suitably renormalized in each model. Surprisingly, the HA-Gō model gives the softest protein, exhibiting much smaller force peaks than all other models after the above renormalization. Clustering to render a structural taxonomy as the protein unfolds showed that the AA, HA-Gō, and Cα-Gō models exhibit a single pathway for early unfolding, which eventually bifurcates repeatedly to multiple branches only after the protein is about half-unfolded. The AWSEM model shows a single dominant unfolding pathway over the whole range of unfolding, in contrast to all other models. TM alignment, clustering analysis, and native contact maps show that the AWSEM pathway has however the most structural similarity to the AA model at high nativeness, but the least structural similarity to the AA model at low nativeness. In comparison to the AA model, the sequence of native contact breakage is best predicted by the HA-Gō model. All models consistently predict a similar unfolding mechanism for early force-induced unfolding events, but diverge in their predictions for late stage unfolding events when the protein is more significantly disordered.

Leaf fossils of Banksia (Proteaceae) from New Zealand: An Australian abroad.

PubMed

Carpenter, Raymond J; Jordan, Gregory J; Lee, Daphne E; Hill, Robert S

2010-02-01

Fossils can shed new light on plant biogeography and phylogeny. Pinnately lobed leaves from the Oligo-Miocene Newvale lignite mine, South Island, New Zealand are the first extra-Australian leaf fossils of the charismatic genus Banksia (Proteaceae), and they are assigned to a new species, B. novae-zelandiae. Comparison with extant taxa shows that the fossils are best regarded as an extinct stem relative of Banksia because their available features are either plesiomorphic for the genus (notably, the stomata are superficially placed, not sunken in balloon-like pits as in many extant species) or lack evidence of synapomorphies that would enable them to be placed in the crown group. Banksia novae-zelandiae does, however, exhibit two cuticular features that are unique or highly derived for Banksia. These are rugulate subsidiary cell ornamentation and the presence of complex papillae that extensively cover the abaxial leaf surface. The fossils add to the widespread records of the pinnately lobed leaf form in Banksia in Australia beginning in the late Paleocene. This form is now limited to species confined to sclerophyllous heathlands of Mediterranean climate in southwestern Australia. Banksia novae-zelandiae could be part of a lineage that had a long history in New Zealand, perhaps dating to the early Paleogene.

Carbon gain and bud physiology in Populus tremuloides and Betula papyrifera grown under long-term exposure to elevated concentrations of CO2 and O3.

PubMed

Riikonen, Johanna; Kets, Katre; Darbah, Joseph; Oksanen, Elina; Sober, Anu; Vapaavuori, Elina; Kubiske, Mark E; Nelson, Neil; Karnosky, David F

2008-02-01

Paper birch (Betula papyrifera Marsh.) and three trembling aspen clones (Populus tremuloides Michx.) were studied to determine if alterations in carbon gain in response to an elevated concentration of CO(2) ([CO(2)]) or O(3) ([O(3)]) or a combination of both affected bud size and carbohydrate composition in autumn, and early leaf development in the following spring. The trees were measured for gas exchange, leaf size, date of leaf abscission, size and biochemical characteristics of the overwintering buds and early leaf development during the 8th-9th year of free-air CO(2) and O(3) exposure at the Aspen FACE site located near Rhinelander, WI. Net photosynthesis was enhanced 49-73% by elevated [CO(2)], and decreased 13-30% by elevated [O(3)]. Elevated [CO(2)] delayed, and elevated [O(3)] tended to accelerate, leaf abscission in autumn. Elevated [CO(2)] increased the ratio of monosaccharides to di- and oligosaccharides in aspen buds, which may indicate a lag in cold acclimation. The total carbon concentration in overwintering buds was unaffected by the treatments, although elevated [O(3)] decreased the amount of starch by 16% in birch buds, and reduced the size of aspen buds, which may be related to the delayed leaf development in aspen during the spring. Elevated [CO(2)] generally ameliorated the effects of elevated [O(3)]. Our results show that both elevated [CO(2)] and elevated [O(3)] have the potential to alter carbon metabolism of overwintering buds. These changes may cause carry-over effects during the next growing season.

Structure of a AAA+ unfoldase in the process of unfolding substrate

PubMed Central

Ripstein, Zev A; Huang, Rui; Augustyniak, Rafal; Kay, Lewis E; Rubinstein, John L

2017-01-01

AAA+ unfoldases are thought to unfold substrate through the central pore of their hexameric structures, but how this process occurs is not known. VAT, the Thermoplasma acidophilum homologue of eukaryotic CDC48/p97, works in conjunction with the proteasome to degrade misfolded or damaged proteins. We show that in the presence of ATP, VAT with its regulatory N-terminal domains removed unfolds other VAT complexes as substrate. We captured images of this transient process by electron cryomicroscopy (cryo-EM) to reveal the structure of the substrate-bound intermediate. Substrate binding breaks the six-fold symmetry of the complex, allowing five of the six VAT subunits to constrict into a tight helix that grips an ~80 Å stretch of unfolded protein. The structure suggests a processive hand-over-hand unfolding mechanism, where each VAT subunit releases the substrate in turn before re-engaging further along the target protein, thereby unfolding it. DOI: http://dx.doi.org/10.7554/eLife.25754.001 PMID:28390173

Conformational dynamics of a protein in the folded and the unfolded state

NASA Astrophysics Data System (ADS)

Fitter, Jörg

2003-08-01

In a quasielastic neutron scattering experiment, the picosecond dynamics of α-amylase was investigated for the folded and the unfolded state of the protein. In order to ensure a reasonable interpretation of the internal protein dynamics, the protein was measured in D 2O-buffer solution. The much higher structural flexibility of the pH induced unfolded state as compared to the native folded state was quantified using a simple analytical model, describing a local diffusion inside a sphere. In terms of this model the conformational volume, which is explored mainly by confined protein side-chain movements, is parameterized by the radius of a sphere (folded state, r=1.2 Å; unfolded state, 1.8 Å). Differences in conformational dynamics between the folded and the unfolded state of a protein are of fundamental interest in the field of protein science, because they are assumed to play an important role for the thermodynamics of folding/unfolding transition and for protein stability.

Common features in the unfolding and misfolding of PDZ domains and beyond: the modulatory effect of domain swapping and extra-elements.

PubMed

Murciano-Calles, Javier; Güell-Bosch, Jofre; Villegas, Sandra; Martinez, Jose C

2016-01-12

PDZ domains are protein-protein interaction modules sharing the same structural arrangement. To discern whether they display common features in their unfolding/misfolding behaviour we have analyzed in this work the unfolding thermodynamics, together with the misfolding kinetics, of the PDZ fold using three archetypical examples: the second and third PDZ domains of the PSD95 protein and the Erbin PDZ domain. Results showed that all domains passed through a common intermediate, which populated upon unfolding, and that this in turn drove the misfolding towards worm-like fibrillar structures. Thus, the unfolding/misfolding behaviour appears to be shared within these domains. We have also analyzed how this landscape can be modified upon the inclusion of extra-elements, as it is in the nNOS PDZ domain, or the organization of swapped species, as happens in the second PDZ domain of the ZO2 protein. Although the intermediates still formed upon thermal unfolding, the misfolding was prevented to varying degrees.

Anomalous Diffusion Reports on the Interaction of Misfolded Proteins with the Quality Control Machinery in the Endoplasmic Reticulum

PubMed Central

Malchus, Nina; Weiss, Matthias

2010-01-01

A multitude of transmembrane proteins enters the endoplasmic reticulum (ER) as unfolded polypeptide chains. During their folding process, they interact repetitively with the ER's quality control machinery. Here, we have used fluorescence correlation spectroscopy to probe these interactions for a prototypical transmembrane protein, VSVG ts045, in vivo. While both folded and unfolded VSVG ts045 showed anomalous diffusion, the unfolded protein had a significantly stronger anomaly. This difference subsided when unfolded VSVG ts045 was in a complex with its chaperone calnexin, or when a mutant form of VSVG ts045 with only one glycan was used. Our experimental data and accompanying simulations suggest that the folding sensor of the quality control (UGT1) oligomerizes unfolded VSVG ts045, leading to a more anomalous/obstructed diffusion. In contrast, calnexin dissolves the oligomers, rendering unfolded VSVG ts045 more mobile, and hence prevents poisoning of the ER. PMID:20713018

Protein unfolding as a switch from self-recognition to high-affinity client binding

PubMed Central

Groitl, Bastian; Horowitz, Scott; Makepeace, Karl A. T.; Petrotchenko, Evgeniy V.; Borchers, Christoph H.; Reichmann, Dana; Bardwell, James C. A.; Jakob, Ursula

2016-01-01

Stress-specific activation of the chaperone Hsp33 requires the unfolding of a central linker region. This activation mechanism suggests an intriguing functional relationship between the chaperone's own partial unfolding and its ability to bind other partially folded client proteins. However, identifying where Hsp33 binds its clients has remained a major gap in our understanding of Hsp33's working mechanism. By using site-specific Fluorine-19 nuclear magnetic resonance experiments guided by in vivo crosslinking studies, we now reveal that the partial unfolding of Hsp33's linker region facilitates client binding to an amphipathic docking surface on Hsp33. Furthermore, our results provide experimental evidence for the direct involvement of conditionally disordered regions in unfolded protein binding. The observed structural similarities between Hsp33's own metastable linker region and client proteins present a possible model for how Hsp33 uses protein unfolding as a switch from self-recognition to high-affinity client binding. PMID:26787517

Determination of thermodynamics and kinetics of RNA reactions by force

PubMed Central

Tinoco, Ignacio; Li, Pan T. X.; Bustamante, Carlos

2008-01-01

Single-molecule methods have made it possible to apply force to an individual RNA molecule. Two beads are attached to the RNA; one is on a micropipette, the other is in a laser trap. The force on the RNA and the distance between the beads are measured. Force can change the equilibrium and the rate of any reaction in which the product has a different extension from the reactant. This review describes use of laser tweezers to measure thermodynamics and kinetics of unfolding/refolding RNA. For a reversible reaction the work directly provides the free energy; for irreversible reactions the free energy is obtained from the distribution of work values. The rate constants for the folding and unfolding reactions can be measured by several methods. The effect of pulling rate on the distribution of force-unfolding values leads to rate constants for unfolding. Hopping of the RNA between folded and unfolded states at constant force provides both unfolding and folding rates. Force-jumps and force-drops, similar to the temperature jump method, provide direct measurement of reaction rates over a wide range of forces. The advantages of applying force and using single-molecule methods are discussed. These methods, for example, allow reactions to be studied in non-denaturing solvents at physiological temperatures; they also simplify analysis of kinetic mechanisms because only one intermediate at a time is present. Unfolding of RNA in biological cells by helicases, or ribosomes, has similarities to unfolding by force. PMID:17040613

Force-Induced Unfolding of Fibronectin in the Extracellular Matrix of Living Cells

PubMed Central

Smith, Michael L; Gourdon, Delphine; Little, William C; Kubow, Kristopher E; Eguiluz, R. Andresen; Luna-Morris, Sheila; Vogel, Viola

2007-01-01

Whether mechanically unfolded fibronectin (Fn) is present within native extracellular matrix fibrils is controversial. Fn extensibility under the influence of cell traction forces has been proposed to originate either from the force-induced lengthening of an initially compact, folded quaternary structure as is found in solution (quaternary structure model, where the dimeric arms of Fn cross each other), or from the force-induced unfolding of type III modules (unfolding model). Clarification of this issue is central to our understanding of the structural arrangement of Fn within fibrils, the mechanism of fibrillogenesis, and whether cryptic sites, which are exposed by partial protein unfolding, can be exposed by cell-derived force. In order to differentiate between these two models, two fluorescence resonance energy transfer schemes to label plasma Fn were applied, with sensitivity to either compact-to-extended conformation (arm separation) without loss of secondary structure or compact-to-unfolded conformation. Fluorescence resonance energy transfer studies revealed that a significant fraction of fibrillar Fn within a three-dimensional human fibroblast matrix is partially unfolded. Complete relaxation of Fn fibrils led to a refolding of Fn. The compactly folded quaternary structure with crossed Fn arms, however, was never detected within extracellular matrix fibrils. We conclude that the resting state of Fn fibrils does not contain Fn molecules with crossed-over arms, and that the several-fold extensibility of Fn fibrils involves the unfolding of type III modules. This could imply that Fn might play a significant role in mechanotransduction processes. PMID:17914904

How long does it take to equilibrate the unfolded state of a protein?

PubMed Central

Levy, Ronald M; Dai, Wei; Deng, Nan-Jie; Makarov, Dmitrii E

2013-01-01

How long does it take to equilibrate the unfolded state of a protein? The answer to this question has important implications for our understanding of why many small proteins fold with two state kinetics. When the equilibration within the unfolded state U is much faster than the folding, the folding kinetics will be two state even if there are many folding pathways with different barriers. Yet the mean first passage times (MFPTs) between different regions of the unfolded state can be much longer than the folding time. This seems to imply that the equilibration within U is much slower than the folding. In this communication we resolve this paradox. We present a formula for estimating the time to equilibrate the unfolded state of a protein. We also present a formula for the MFPT to any state within U, which is proportional to the average lifetime of that state divided by the state population. This relation is valid when the equilibration within U is very fast as compared with folding as it often is for small proteins. To illustrate the concepts, we apply the formulas to estimate the time to equilibrate the unfolded state of Trp-cage and MFPTs within the unfolded state based on a Markov State Model using an ultra-long 208 microsecond trajectory of the miniprotein to parameterize the model. The time to equilibrate the unfolded state of Trp-cage is ∼100 ns while the typical MFPTs within U are tens of microseconds or longer. PMID:23963761

Increased Atmospheric SO2 Detected from Changes in Leaf Physiognomy across the Triassic–Jurassic Boundary Interval of East Greenland

PubMed Central

Bacon, Karen L.; Belcher, Claire M.; Haworth, Matthew; McElwain, Jennifer C.

2013-01-01

The Triassic–Jurassic boundary (Tr–J; ∼201 Ma) is marked by a doubling in the concentration of atmospheric CO2, rising temperatures, and ecosystem instability. This appears to have been driven by a major perturbation in the global carbon cycle due to massive volcanism in the Central Atlantic Magmatic Province. It is hypothesized that this volcanism also likely delivered sulphur dioxide (SO2) to the atmosphere. The role that SO2 may have played in leading to ecosystem instability at the time has not received much attention. To date, little direct evidence has been presented from the fossil record capable of implicating SO2 as a cause of plant extinctions at this time. In order to address this, we performed a physiognomic leaf analysis on well-preserved fossil leaves, including Ginkgoales, bennettites, and conifers from nine plant beds that span the Tr–J boundary at Astartekløft, East Greenland. The physiognomic responses of fossil taxa were compared to the leaf size and shape variations observed in nearest living equivalent taxa exposed to simulated palaeoatmospheric treatments in controlled environment chambers. The modern taxa showed a statistically significant increase in leaf roundness when fumigated with SO2. A similar increase in leaf roundness was also observed in the Tr–J fossil taxa immediately prior to a sudden decrease in their relative abundances at Astartekløft. This research reveals that increases in atmospheric SO2 can likely be traced in the fossil record by analyzing physiognomic changes in fossil leaves. A pattern of relative abundance decline following increased leaf roundness for all six fossil taxa investigated supports the hypothesis that SO2 had a significant role in Tr–J plant extinctions. This finding highlights that the role of SO2 in plant biodiversity declines across other major geological boundaries coinciding with global scale volcanism should be further explored using leaf physiognomy. PMID:23593262

Evidence of Physiological Decoupling from Grassland Ecosystem Drivers by an Encroaching Woody Shrub

PubMed Central

Nippert, Jesse B.; Ocheltree, Troy W.; Orozco, Graciela L.; Ratajczak, Zak; Ling, Bohua; Skibbe, Adam M.

2013-01-01

Shrub encroachment of grasslands is a transformative ecological process by which native woody species increase in cover and frequency and replace the herbaceous community. Mechanisms of encroachment are typically assessed using temporal data or experimental manipulations, with few large spatial assessments of shrub physiology. In a mesic grassland in North America, we measured inter- and intra-annual variability in leaf δ13C in Cornus drummondii across a grassland landscape with varying fire frequency, presence of large grazers and topographic variability. This assessment of changes in individual shrub physiology is the largest spatial and temporal assessment recorded to date. Despite a doubling of annual rainfall (in 2008 versus 2011), leaf δ13C was statistically similar among and within years from 2008-11 (range of −28 to −27‰). A topography*grazing interaction was present, with higher leaf δ13C in locations that typically have more bare soil and higher sensible heat in the growing season (upland topographic positions and grazed grasslands). Leaf δ13C from slopes varied among grazing contrasts, with upland and slope leaf δ13C more similar in ungrazed locations, while slopes and lowlands were more similar in grazed locations. In 2011, canopy greenness (normalized difference vegetation index – NDVI) was assessed at the centroid of individual shrubs using high-resolution hyperspectral imagery. Canopy greenness was highest mid-summer, likely reflecting temporal periods when C assimilation rates were highest. Similar to patterns seen in leaf δ13C, NDVI was highest in locations that typically experience lowest sensible heat (lowlands and ungrazed). The ability of Cornus drummondii to decouple leaf physiological responses from climate variability and fire frequency is a likely contributor to the increase in cover and frequency of this shrub species in mesic grassland and may be generalizable to other grasslands undergoing woody encroachment. PMID:24339950

Phenological series in Bologna (Northern Italy): temporal trends and spatial pattern of greening

NASA Astrophysics Data System (ADS)

Puppi, Giovanna; Zanotti, Anna Letizia

2010-05-01

In Europe, in the last thirty years, the onset of spring has advanced (Estrella et al. 2009 Clim Res. 39). However the phenological shifts vary from one country to another and long time series are scanty in Southern Europe (Peňuelas et. al. 2002, Glob. Change Biol.; Črepinšek et Kajfež 2003, Ann.Ser.Hist.Nat.13): for this reason we analyzed the data on leafing of some woody species recorded during the last 3 decades (1977-2009) in the city of Bologna (Emilia-Romagna, Northern Italy) and in several hillside stations in the neighbourhood of the town. As BGS index, the average day of "first leaf unfolding" (GFI=V4, BBCH=11) of a group of common woody species was chosen: the BGS days were analysed in relation with time and temperature changes. The BGS happens on average at the end of March. The species observed display a negative trend along the period (about 2 days of advance per decade) and the BGS days show a significant correlation with the mean temperature of the period January-March (about 3 and a half days of earlier start per degree of increasing temperature). Since an increase in winter temperatures has been predicted in the Emilia-Romagna region in the next decades (Tomozeiu et al. 2007, Theor. Appl. Climatol. 90), the growing season in our region is probably going to extend. Phenological spatial patterns of the neighbourhood of the city are also analysed: greening generally starts on southfacing hills between 100 and 300 m asl.

A Latent Class Unfolding Model for Analyzing Single Stimulus Preference Ratings.

ERIC Educational Resources Information Center

De Soete, Geert; Heiser, Willem J.

1993-01-01

A latent class unfolding model is developed for single stimulus preference ratings. One advantage is the possibility of testing the spatial unfolding model against the unconstrained latent class model for rating data. The model is applied to data about party preferences of members of the Dutch parliament. (SLD)

XBP1, Unfolded Protein Response, and Endocrine Responsiveness

DTIC Science & Technology

2011-05-01

initially modeled in yeast cells (21, 22). Components of the unfolded protein response (UPR) are also conserved across species and these include...response in tumors. Mol Cancer Res 2005;3:597–605. 38. Sriburi R, Jackowski S, Mori K, Brewer JW. XBP1: a link between the unfolded protein response

Unfolding Case-Based Practicum Curriculum Infusing Crisis, Trauma, and Disaster Preparation

ERIC Educational Resources Information Center

Greene, Catie A.; Williams, Amy E.; Harris, Pamela N.; Travis, Sterling P.; Kim, Sharon Y.

2016-01-01

The authors evaluated an unfolding case-based approach to a practicum in counseling course infusing crisis, trauma, and disaster preparation for changes in students' crisis self-efficacy across a semester. The course, informed by constructivist-developmental pedagogy and centered on the unfolding case, resulted in significant increases in…

Unfolding the prompt gamma ray spectra measured in a Lanthanum Bromide detector using GRAVEL method

NASA Astrophysics Data System (ADS)

De, S.; Thomas, R. G.; Rout, P. C.; Suryanarayana, S. V.; Nayak, B. K.; Saxena, A.

2018-02-01

Prompt fission Upsilon -ray energy spectra in spontaneous fission of 252Cf has been measured using a 6'' LaBr3(Ce) detector. Unfolding of the measured Upsilon -ray energy spectra has been carried out using GRAVEL method. The response matrix of the detector has been simulated using GEANT4 and the unfolding of Upsilon -ray energy spectra for 60Co and 137Cs sources have been validated. This unfolding technique has then been applied to the prompt gamma spectra obtained from the spontaneous fission of 252Cf.

Molecular Dynamics Simulations of the Temperature Induced Unfolding of Crambin Follow the Arrhenius Equation.

PubMed

Dalby, Andrew; Shamsir, Mohd Shahir

2015-01-01

Molecular dynamics simulations have been used extensively to model the folding and unfolding of proteins. The rates of folding and unfolding should follow the Arrhenius equation over a limited range of temperatures. This study shows that molecular dynamic simulations of the unfolding of crambin between 500K and 560K do follow the Arrhenius equation. They also show that while there is a large amount of variation between the simulations the average values for the rate show a very high degree of correlation.

Molecular Dynamics Simulations of the Temperature Induced Unfolding of Crambin Follow the Arrhenius Equation.

PubMed Central

Dalby, Andrew; Shamsir, Mohd Shahir

2015-01-01

Molecular dynamics simulations have been used extensively to model the folding and unfolding of proteins. The rates of folding and unfolding should follow the Arrhenius equation over a limited range of temperatures. This study shows that molecular dynamic simulations of the unfolding of crambin between 500K and 560K do follow the Arrhenius equation. They also show that while there is a large amount of variation between the simulations the average values for the rate show a very high degree of correlation. PMID:26539292

Proof-of-principle to unfold an angle-energy dependent source from forward and adjoint calculations

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

Pace, J.V. III

For many years there has existed a discrepancy between the measured and calculated responses from the Little Boy weapon in Hiroshima. A myriad of solutions have been proposed, but to no avail. If one can rationalize to himself that it does not really matter exactly what happened with the weapon when it exploded, and if sufficient information exist about the measurements, one should be able to unfold the source. Moreover, if a source can be unfolded in a controlled environment, then it should be possible to unfold a more complicated source, for example, the Little Boy source. This report recordsmore » the findings of a proof-of-principle test to unfold a source in the controlled environment.« less

Unfolding mechanism of lysozyme in various urea solutions: Insights from fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Chen, Bang; Zhang, Hongjia; Xi, Wenying; Zhao, Liqing; Liang, Li; Chen, Yantao

2014-11-01

Fluorescence spectroscopic technique is very popular in exploring the folding/unfolding process of proteins. In this paper, unfolding process of hen egg-white lysozyme was investigated in various denaturing solutions. Firstly, polymer solution theory was employed to comprehend the dependence of fluorescence quenching effect on protein concentration, and dynamic contact concentration was suggested as a critical value for related fluorescence experiment. Secondly, it was found that urea alone could not completely unfold lysozyme but did when together with DTT or HCl. Lysozyme was destabilized in concentrated urea solution, but still could maintain its spatial structure. Phase diagram of fluorescence intensities revealed that HCl could enhance the denaturing capacity of urea, resulting in the emergence of intermediate state in the thermodynamic unfolding process of lysozyme.

Unfolding energy spectra of double-periodicity two-dimensional systems: Twisted bilayer graphene and MoS2 on graphene

NASA Astrophysics Data System (ADS)

Matsushita, Yu-ichiro; Nishi, Hirofumi; Iwata, Jun-ichi; Kosugi, Taichi; Oshiyama, Atsushi

2018-01-01

We propose an unfolding scheme to analyze energy spectra of complex large-scale systems which are inherently of double periodicity on the basis of the density-functional theory. Applying our method to a twisted bilayer graphene (tBLG) and a stack of monolayer MoS2 on graphene (MoS2/graphene) as examples, we first show that the conventional unfolding scheme in the past using a single primitive-cell representation causes serious problems in analyses of the energy spectra. We then introduce our multispace representation scheme in the unfolding method and clarify its validity. Velocity renormalization of Dirac electrons in tBLG and mini gaps of Dirac cones in MoS2/graphene are elucidated in the present unfolding scheme.

Nonparametric density estimation and optimal bandwidth selection for protein unfolding and unbinding data

NASA Astrophysics Data System (ADS)

Bura, E.; Zhmurov, A.; Barsegov, V.

2009-01-01

Dynamic force spectroscopy and steered molecular simulations have become powerful tools for analyzing the mechanical properties of proteins, and the strength of protein-protein complexes and aggregates. Probability density functions of the unfolding forces and unfolding times for proteins, and rupture forces and bond lifetimes for protein-protein complexes allow quantification of the forced unfolding and unbinding transitions, and mapping the biomolecular free energy landscape. The inference of the unknown probability distribution functions from the experimental and simulated forced unfolding and unbinding data, as well as the assessment of analytically tractable models of the protein unfolding and unbinding requires the use of a bandwidth. The choice of this quantity is typically subjective as it draws heavily on the investigator's intuition and past experience. We describe several approaches for selecting the "optimal bandwidth" for nonparametric density estimators, such as the traditionally used histogram and the more advanced kernel density estimators. The performance of these methods is tested on unimodal and multimodal skewed, long-tailed distributed data, as typically observed in force spectroscopy experiments and in molecular pulling simulations. The results of these studies can serve as a guideline for selecting the optimal bandwidth to resolve the underlying distributions from the forced unfolding and unbinding data for proteins.

Multistage unfolding of an SH3 domain: an initial urea-filled dry molten globule precedes a wet molten globule with non-native structure.

PubMed

Dasgupta, Amrita; Udgaonkar, Jayant B; Das, Payel

2014-06-19

The unfolding of the SH3 domain of the PI3 kinase in aqueous urea has been studied using a synergistic experiment-simulation approach. The experimental observation of a transient wet molten globule intermediate, IU, with an unusual non-native burial of the sole Trp residue, W53, provides the benchmark for the unfolding simulations performed (eight in total, each at least 0.5 μs long). The simulations reveal that the partially unfolded IU ensemble is preceded by an early native-like molten globule intermediate ensemble I*. In the very initial stage of unfolding, dry globule conformations with the protein core filled with urea instead of water are transiently observed within the I* ensemble. Water penetration into the urea-filled core of dry globule conformations is frequently accompanied by very transient burial of W53. Later during gradual unfolding, W53 is seen to again become transiently buried in the IU ensemble for a much longer time. In the structurally heterogeneous IU ensemble, conformational flexibility of the C-terminal β-strands enables W53 burial by the formation of non-native, tertiary contacts with hydrophobic residues, which could serve to protect the protein from aggregation during unfolding.

Characterization of two distinct beta2-microglobulin unfolding intermediates that may lead to amyloid fibrils of different morphology.

PubMed

Armen, Roger S; Daggett, Valerie

2005-12-13

The self-assembly of beta(2)-microglobulin into fibrils leads to dialysis-related amyloidosis. pH-mediated partial unfolding is required for the formation of the amyloidogenic intermediate that then self-assembles into amyloid fibrils. Two partially folded intermediates of beta(2)-microglobulin have been identified experimentally and linked to the formation of fibrils of distinct morphology, yet it remains difficult to characterize these partially unfolded states at high resolution using experimental approaches. Consequently, we have performed molecular dynamics simulations at neutral and low pH to determine the structures of these partially unfolded amyloidogenic intermediates. In the low-pH simulations, we observed the formation of alpha-sheet structure, which was first proposed by Pauling and Corey. Multiple simulations were performed, and two distinct intermediate state ensembles were identified that may account for the different fibril morphologies. The predominant early unfolding intermediate was nativelike in structure, in agreement with previous NMR studies. The late unfolding intermediate was significantly disordered, but it maintained an extended elongated structure, with hydrophobic clusters and residual alpha-extended chain strands in specific regions of the sequence that map to amyloidogenic peptides. We propose that the formation of alpha-sheet facilitates self-assembly into partially unfolded prefibrillar amyloidogenic intermediates.

Branches of Triangulated Origami Near the Unfolded State

NASA Astrophysics Data System (ADS)

Chen, Bryan Gin-ge; Santangelo, Christian D.

2018-01-01

Origami structures are characterized by a network of folds and vertices joining unbendable plates. For applications to mechanical design and self-folding structures, it is essential to understand the interplay between the set of folds in the unfolded origami and the possible 3D folded configurations. When deforming a structure that has been folded, one can often linearize the geometric constraints, but the degeneracy of the unfolded state makes a linear approach impossible there. We derive a theory for the second-order infinitesimal rigidity of an initially unfolded triangulated origami structure and use it to study the set of nearly unfolded configurations of origami with four boundary vertices. We find that locally, this set consists of a number of distinct "branches" which intersect at the unfolded state, and that the number of these branches is exponential in the number of vertices. We find numerical and analytical evidence that suggests that the branches are characterized by choosing each internal vertex to either "pop up" or "pop down." The large number of pathways along which one can fold an initially unfolded origami structure strongly indicates that a generic structure is likely to become trapped in a "misfolded" state. Thus, new techniques for creating self-folding origami are likely necessary; controlling the popping state of the vertices may be one possibility.

Studies on the Dissociation and Urea-Induced Unfolding of FtsZ Support the Dimer Nucleus Polymerization Mechanism

PubMed Central

Montecinos-Franjola, Felipe; Ross, Justin A.; Sánchez, Susana A.; Brunet, Juan E.; Lagos, Rosalba; Jameson, David M.; Monasterio, Octavio

2012-01-01

FtsZ is a major protein in bacterial cytokinesis that polymerizes into single filaments. A dimer has been proposed to be the nucleating species in FtsZ polymerization. To investigate the influence of the self-assembly of FtsZ on its unfolding pathway, we characterized its oligomerization and unfolding thermodynamics. We studied the assembly using size-exclusion chromatography and fluorescence spectroscopy, and the unfolding using circular dichroism and two-photon fluorescence correlation spectroscopy. The chromatographic analysis demonstrated the presence of monomers, dimers, and tetramers with populations dependent on protein concentration. Dilution experiments using fluorescent conjugates revealed dimer-to-monomer and tetramer-to-dimer dissociation constants in the micromolar range. Measurements of fluorescence lifetimes and rotational correlation times of the conjugates supported the presence of tetramers at high protein concentrations and monomers at low protein concentrations. The unfolding study demonstrated that the three-state unfolding of FtsZ was due to the mainly dimeric state of the protein, and that the monomer unfolds through a two-state mechanism. The monomer-to-dimer equilibrium characterized here (Kd = 9 μM) indicates a significant fraction (∼10%) of stable dimers at the critical concentration for polymerization, supporting a role of the dimeric species in the first steps of FtsZ polymerization. PMID:22824282

Thermal, Chemical and pH Induced Denaturation of a Multimeric β-Galactosidase Reveals Multiple Unfolding Pathways

PubMed Central

Kishore, Devesh; Kundu, Suman; Kayastha, Arvind M.

2012-01-01

Background In this case study, we analysed the properties of unfolded states and pathways leading to complete denaturation of a multimeric chick pea β-galactosidase (CpGAL), as obtained from treatment with guanidium hydrochloride, urea, elevated temperature and extreme pH. Methodology/Principal Findings CpGAL, a heterodimeric protein with native molecular mass of 85 kDa, belongs to α+β class of protein. The conformational stability and thermodynamic parameters of CpGAL unfolding in different states were estimated and interpreted using circular dichroism and fluorescence spectroscopic measurements. The enzyme was found to be structurally and functionally stable in the entire pH range and upto 50°C temperature. Further increase in temperature induces unfolding followed by aggregation. Chemical induced denaturation was found to be cooperative and transitions were irreversible, non-coincidental and sigmoidal. Free energy of protein unfolding (ΔG0) and unfolding constant (Kobs) were also calculated for chemically denatured CpGAL. Significance The protein seems to use different pathways for unfolding in different environments and is a classical example of how the environment dictates the path a protein might take to fold while its amino acid sequence only defines its final three-dimensional conformation. The knowledge accumulated could be of immense biotechnological significance as well. PMID:23185611

First Passage Times, Lifetimes, and Relaxation Times of Unfolded Proteins

NASA Astrophysics Data System (ADS)

Dai, Wei; Sengupta, Anirvan M.; Levy, Ronald M.

2015-07-01

The dynamics of proteins in the unfolded state can be quantified in computer simulations by calculating a spectrum of relaxation times which describes the time scales over which the population fluctuations decay to equilibrium. If the unfolded state space is discretized, we can evaluate the relaxation time of each state. We derive a simple relation that shows the mean first passage time to any state is equal to the relaxation time of that state divided by the equilibrium population. This explains why mean first passage times from state to state within the unfolded ensemble can be very long but the energy landscape can still be smooth (minimally frustrated). In fact, when the folding kinetics is two-state, all of the unfolded state relaxation times within the unfolded free energy basin are faster than the folding time. This result supports the well-established funnel energy landscape picture and resolves an apparent contradiction between this model and the recently proposed kinetic hub model of protein folding. We validate these concepts by analyzing a Markov state model of the kinetics in the unfolded state and folding of the miniprotein NTL9 (where NTL9 is the N -terminal domain of the ribosomal protein L9), constructed from a 2.9 ms simulation provided by D. E. Shaw Research.

First Passage Times, Lifetimes, and Relaxation Times of Unfolded Proteins.

PubMed

Dai, Wei; Sengupta, Anirvan M; Levy, Ronald M

2015-07-24

The dynamics of proteins in the unfolded state can be quantified in computer simulations by calculating a spectrum of relaxation times which describes the time scales over which the population fluctuations decay to equilibrium. If the unfolded state space is discretized, we can evaluate the relaxation time of each state. We derive a simple relation that shows the mean first passage time to any state is equal to the relaxation time of that state divided by the equilibrium population. This explains why mean first passage times from state to state within the unfolded ensemble can be very long but the energy landscape can still be smooth (minimally frustrated). In fact, when the folding kinetics is two-state, all of the unfolded state relaxation times within the unfolded free energy basin are faster than the folding time. This result supports the well-established funnel energy landscape picture and resolves an apparent contradiction between this model and the recently proposed kinetic hub model of protein folding. We validate these concepts by analyzing a Markov state model of the kinetics in the unfolded state and folding of the miniprotein NTL9 (where NTL9 is the N-terminal domain of the ribosomal protein L9), constructed from a 2.9 ms simulation provided by D. E. Shaw Research.

Isothermal chemical denaturation of large proteins: Path-dependence and irreversibility.

PubMed

Wafer, Lucas; Kloczewiak, Marek; Polleck, Sharon M; Luo, Yin

2017-12-15

State functions (e.g., ΔG) are path independent and quantitatively describe the equilibrium states of a thermodynamic system. Isothermal chemical denaturation (ICD) is often used to extrapolate state function parameters for protein unfolding in native buffer conditions. The approach is prudent when the unfolding/refolding processes are path independent and reversible, but may lead to erroneous results if the processes are not reversible. The reversibility was demonstrated in several early studies for smaller proteins, but was assumed in some reports for large proteins with complex structures. In this work, the unfolding/refolding of several proteins were systematically studied using an automated ICD instrument. It is shown that: (i) the apparent unfolding mechanism and conformational stability of large proteins can be denaturant-dependent, (ii) equilibration times for large proteins are non-trivial and may introduce significant error into calculations of ΔG, (iii) fluorescence emission spectroscopy may not correspond to other methods, such as circular dichroism, when used to measure protein unfolding, and (iv) irreversible unfolding and hysteresis can occur in the absence of aggregation. These results suggest that thorough confirmation of the state functions by, for example, performing refolding experiments or using additional denaturants, is needed when quantitatively studying the thermodynamics of protein unfolding using ICD. Copyright © 2017 Elsevier Inc. All rights reserved.

High Temperature Unfolding and Low Temperature Refolding Pathway of Chymotrypsin Inhibitor 2 Using Molecular Dynamics Simulation

NASA Astrophysics Data System (ADS)

Malau, N. D.; Sumaryada, T.

2016-01-01

The mechanism that explains the unfolding/refolding process of the protein is still a major problem that has not been fully understood. In this paper we present our study on the unfolding and refolding pathway of Chymotrypsin Inhibitor 2 (CI2) protein through a molecular dynamics simulation technique. The high temperature unfolding simulation were performed at 500 K for 35 ns. While the low temperature refolding simulation performed at 200 K for 35 ns. The unfolding and refolding pathway of protein were analysed by looking at the dynamics of root mean squared deviation (RMSD) and secondary structure profiles. The signatures of unfolding were observed from significant increase of RMSD within the time span of 10 ns to 35 ns. For the refolding process, the initial structure was prepared from the structure of unfolding protein at t=15 ns and T=500 K. Analysis have shown that some of the secondary structures of CI2 protein that have been damaged at high temperature can be refolded back to its initial structure at low temperature simulation. Our results suggest that most of α-helix structure of CI2 protein can be refolded back to its initial state, while only half beta-sheet structure can be reformed.

Engineered Bi-Histidine Metal Chelation Sites Map the Structure of the Mechanical Unfolding Transition State of an Elastomeric Protein Domain GB1

PubMed Central

Shen, Tao; Cao, Yi; Zhuang, Shulin; Li, Hongbin

2012-01-01

Determining the structure of the transition state is critical for elucidating the mechanism behind how proteins fold and unfold. Due to its high free energy, however, the transition state generally cannot be trapped and studied directly using traditional structural biology methods. Thus, characterizing the structure of the transition state that occurs as proteins fold and unfold remains a major challenge. Here, we report a novel (to our knowledge) method that uses engineered bi-histidine (bi-His) metal-binding sites to directly map the structure of the mechanical unfolding transition state of proteins. This method is adapted from the traditional ψ-value analysis, which uses engineered bi-His metal chelation sites to probe chemical (un)folding transition-state structure. The ϕM2+U-value is defined as ΔΔG‡-N/ΔΔGU-N, which is the energetic effects of metal chelation by the bi-His site on the unfolding energy barrier (ΔG‡-N) relative to its thermodynamic stability (ΔGU-N) and can be used to obtain information about the transition state in the mutational site. As a proof of principle, we used the small protein GB1 as a model system and set out to map its mechanical unfolding transition-state structure. Using single-molecule atomic force microscopy and spectrofluorimetry, we directly quantified the effect of divalent metal ion binding on the mechanical unfolding free energy and thermodynamic stability of GB1, which allowed us to quantify ϕM2+U-values for different sites in GB1. Our results enabled us to map the structure of the mechanical unfolding transition state of GB1. Within GB1’s mechanical unfolding transition state, the interface between force-bearing β-strands 1 and 4 is largely disrupted, and the first β-hairpin is partially disordered while the second β-hairpin and the α-helix remain structured. Our results demonstrate the unique application of ψ-value analysis in elucidating the structure of the transition state that occurs during the mechanical unfolding process, offering a potentially powerful new method for investigating the design of novel elastomeric proteins. PMID:22947942

Engineered bi-histidine metal chelation sites map the structure of the mechanical unfolding transition state of an elastomeric protein domain GB1.

PubMed

Shen, Tao; Cao, Yi; Zhuang, Shulin; Li, Hongbin

2012-08-22

Determining the structure of the transition state is critical for elucidating the mechanism behind how proteins fold and unfold. Due to its high free energy, however, the transition state generally cannot be trapped and studied directly using traditional structural biology methods. Thus, characterizing the structure of the transition state that occurs as proteins fold and unfold remains a major challenge. Here, we report a novel (to our knowledge) method that uses engineered bi-histidine (bi-His) metal-binding sites to directly map the structure of the mechanical unfolding transition state of proteins. This method is adapted from the traditional ψ-value analysis, which uses engineered bi-His metal chelation sites to probe chemical (un)folding transition-state structure. The φ(M2+)(U)-value is defined as ΔΔG(‡-N)/ΔΔG(U-N), which is the energetic effects of metal chelation by the bi-His site on the unfolding energy barrier (ΔG(‡-N)) relative to its thermodynamic stability (ΔG(U-N)) and can be used to obtain information about the transition state in the mutational site. As a proof of principle, we used the small protein GB1 as a model system and set out to map its mechanical unfolding transition-state structure. Using single-molecule atomic force microscopy and spectrofluorimetry, we directly quantified the effect of divalent metal ion binding on the mechanical unfolding free energy and thermodynamic stability of GB1, which allowed us to quantify φ(M2+)(U)-values for different sites in GB1. Our results enabled us to map the structure of the mechanical unfolding transition state of GB1. Within GB1's mechanical unfolding transition state, the interface between force-bearing β-strands 1 and 4 is largely disrupted, and the first β-hairpin is partially disordered while the second β-hairpin and the α-helix remain structured. Our results demonstrate the unique application of ψ-value analysis in elucidating the structure of the transition state that occurs during the mechanical unfolding process, offering a potentially powerful new method for investigating the design of novel elastomeric proteins. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Sequential protein unfolding through a carbon nanotube pore

NASA Astrophysics Data System (ADS)

Xu, Zhonghe; Zhang, Shuang; Weber, Jeffrey K.; Luan, Binquan; Zhou, Ruhong; Li, Jingyuan

2016-06-01

An assortment of biological processes, like protein degradation and the transport of proteins across membranes, depend on protein unfolding events mediated by nanopore interfaces. In this work, we exploit fully atomistic simulations of an artificial, CNT-based nanopore to investigate the nature of ubiquitin unfolding. With one end of the protein subjected to an external force, we observe non-canonical unfolding behaviour as ubiquitin is pulled through the pore opening. Secondary structural elements are sequentially detached from the protein and threaded into the nanotube, interestingly, the remaining part maintains native-like characteristics. The constraints of the nanopore interface thus facilitate the formation of stable ``unfoldon'' motifs above the nanotube aperture that can exist in the absence of specific native contacts with the other secondary structure. Destruction of these unfoldons gives rise to distinct force peaks in our simulations, providing us with a sensitive probe for studying the kinetics of serial unfolding events. Our detailed analysis of nanopore-mediated protein unfolding events not only provides insight into how related processes might proceed in the cell, but also serves to deepen our understanding of structural arrangements which form the basis for protein conformational stability.An assortment of biological processes, like protein degradation and the transport of proteins across membranes, depend on protein unfolding events mediated by nanopore interfaces. In this work, we exploit fully atomistic simulations of an artificial, CNT-based nanopore to investigate the nature of ubiquitin unfolding. With one end of the protein subjected to an external force, we observe non-canonical unfolding behaviour as ubiquitin is pulled through the pore opening. Secondary structural elements are sequentially detached from the protein and threaded into the nanotube, interestingly, the remaining part maintains native-like characteristics. The constraints of the nanopore interface thus facilitate the formation of stable ``unfoldon'' motifs above the nanotube aperture that can exist in the absence of specific native contacts with the other secondary structure. Destruction of these unfoldons gives rise to distinct force peaks in our simulations, providing us with a sensitive probe for studying the kinetics of serial unfolding events. Our detailed analysis of nanopore-mediated protein unfolding events not only provides insight into how related processes might proceed in the cell, but also serves to deepen our understanding of structural arrangements which form the basis for protein conformational stability. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr00410e

Predictors of natively unfolded proteins: unanimous consensus score to detect a twilight zone between order and disorder in generic datasets.

PubMed

Deiana, Antonio; Giansanti, Andrea

2010-04-21

Natively unfolded proteins lack a well defined three dimensional structure but have important biological functions, suggesting a re-assignment of the structure-function paradigm. To assess that a given protein is natively unfolded requires laborious experimental investigations, then reliable sequence-only methods for predicting whether a sequence corresponds to a folded or to an unfolded protein are of interest in fundamental and applicative studies. Many proteins have amino acidic compositions compatible both with the folded and unfolded status, and belong to a twilight zone between order and disorder. This makes difficult a dichotomic classification of protein sequences into folded and natively unfolded ones. In this work we propose an operational method to identify proteins belonging to the twilight zone by combining into a consensus score good performing single predictors of folding. In this methodological paper dichotomic folding indexes are considered: hydrophobicity-charge, mean packing, mean pairwise energy, Poodle-W and a new global index, that is called here gVSL2, based on the local disorder predictor VSL2. The performance of these indexes is evaluated on different datasets, in particular on a new dataset composed by 2369 folded and 81 natively unfolded proteins. Poodle-W, gVSL2 and mean pairwise energy have good performance and stability in all the datasets considered and are combined into a strictly unanimous combination score SSU, that leaves proteins unclassified when the consensus of all combined indexes is not reached. The unclassified proteins: i) belong to an overlap region in the vector space of amino acidic compositions occupied by both folded and unfolded proteins; ii) are composed by approximately the same number of order-promoting and disorder-promoting amino acids; iii) have a mean flexibility intermediate between that of folded and that of unfolded proteins. Our results show that proteins unclassified by SSU belong to a twilight zone. Proteins left unclassified by the consensus score SSU have physical properties intermediate between those of folded and those of natively unfolded proteins and their structural properties and evolutionary history are worth to be investigated.

Predictors of natively unfolded proteins: unanimous consensus score to detect a twilight zone between order and disorder in generic datasets

PubMed Central

2010-01-01

Background Natively unfolded proteins lack a well defined three dimensional structure but have important biological functions, suggesting a re-assignment of the structure-function paradigm. To assess that a given protein is natively unfolded requires laborious experimental investigations, then reliable sequence-only methods for predicting whether a sequence corresponds to a folded or to an unfolded protein are of interest in fundamental and applicative studies. Many proteins have amino acidic compositions compatible both with the folded and unfolded status, and belong to a twilight zone between order and disorder. This makes difficult a dichotomic classification of protein sequences into folded and natively unfolded ones. In this work we propose an operational method to identify proteins belonging to the twilight zone by combining into a consensus score good performing single predictors of folding. Results In this methodological paper dichotomic folding indexes are considered: hydrophobicity-charge, mean packing, mean pairwise energy, Poodle-W and a new global index, that is called here gVSL2, based on the local disorder predictor VSL2. The performance of these indexes is evaluated on different datasets, in particular on a new dataset composed by 2369 folded and 81 natively unfolded proteins. Poodle-W, gVSL2 and mean pairwise energy have good performance and stability in all the datasets considered and are combined into a strictly unanimous combination score SSU, that leaves proteins unclassified when the consensus of all combined indexes is not reached. The unclassified proteins: i) belong to an overlap region in the vector space of amino acidic compositions occupied by both folded and unfolded proteins; ii) are composed by approximately the same number of order-promoting and disorder-promoting amino acids; iii) have a mean flexibility intermediate between that of folded and that of unfolded proteins. Conclusions Our results show that proteins unclassified by SSU belong to a twilight zone. Proteins left unclassified by the consensus score SSU have physical properties intermediate between those of folded and those of natively unfolded proteins and their structural properties and evolutionary history are worth to be investigated. PMID:20409339

Maternally inherited architecture in tertiary leaf beetles: paleoichnology of cryptocephaline fecal cases in Dominican and Baltic amber.

PubMed

Chaboo, Caroline S; Engel, Michael S; Chamorro-Lacayo, Maria Lourdes

2009-09-01

Complex ethological adaptations and intraspecific interactions leave few fossil traces. We document three Dominican (20 million years old [myo]) and Baltic (45 myo) amber fossils that exhibit firm evidence of highly integrated interactions between mothers and offspring in the diverse camptosomate lineage of beetles (Chrysomelidae, leaf beetles). As in contemporary species, these hard cases were initially constructed by mothers, then inherited and retained by offspring, which then elaborate this protective domicile with an unusual but economical building material, their feces. The three fossils are classified in the Subfamily Cryptocephalinae; two are classified in the tribe Chlamisini based on morphological evidence-the flattened head lacking a sharp keel and long legs with simple recurved untoothed claws. These diagnostic features are not clearly visible in the third specimen to permit more refined identification. These fossils provide more precise paleontological dating of tribal nodes within the cryptocephaline radiation of leaf beetles. These fossils are the first and earliest evidence of mother-offspring interaction, building behavior, and fecal recycling in Camptosomata beetles and of inheritance of architectural structures in beetles.

Maternally inherited architecture in tertiary leaf beetles: paleoichnology of cryptocephaline fecal cases in Dominican and Baltic amber

NASA Astrophysics Data System (ADS)

Chaboo, Caroline S.; Engel, Michael S.; Chamorro-Lacayo, Maria Lourdes

2009-09-01

Complex ethological adaptations and intraspecific interactions leave few fossil traces. We document three Dominican (20 million years old [myo]) and Baltic (45 myo) amber fossils that exhibit firm evidence of highly integrated interactions between mothers and offspring in the diverse camptosomate lineage of beetles (Chrysomelidae, leaf beetles). As in contemporary species, these hard cases were initially constructed by mothers, then inherited and retained by offspring, which then elaborate this protective domicile with an unusual but economical building material, their feces. The three fossils are classified in the Subfamily Cryptocephalinae; two are classified in the tribe Chlamisini based on morphological evidence—the flattened head lacking a sharp keel and long legs with simple recurved untoothed claws. These diagnostic features are not clearly visible in the third specimen to permit more refined identification. These fossils provide more precise paleontological dating of tribal nodes within the cryptocephaline radiation of leaf beetles. These fossils are the first and earliest evidence of mother-offspring interaction, building behavior, and fecal recycling in Camptosomata beetles and of inheritance of architectural structures in beetles.

Tracking forest canopy dynamics from an automated proximal hyperspectral monitoring system: linking remote sensing observations to leaf level photosynthetic processes

NASA Astrophysics Data System (ADS)

Woodgate, W.; van Gorsel, E.; Hughes, D.; Suarez, L.; Cabello-Leblic, A.; Held, A. A.; Norton, A.; Dempsey, R.

2017-12-01

To better understand the vegetation response to climate extremes we have developed a fully automated hyperspectral and thermal monitoring system installed on a flux tower at a mature Eucalypt forest site - Tumbarumba, Australia. The automated system bridges spatial, spectral and temporal scales between satellite and in situ observations. Here, we have been acquiring high resolution panoramic hyperspectral and thermal images of the forest canopy three times per day since mid-2014.A specific focus of the work to date has been linking light use efficiency (LUE) as measured by the flux tower to remote sensing observations from the leaf, to crown, to canopy scale. Specifically, targeted field campaigns were conducted in 2016 to establish the interrelationship between structure, function, and spectra. At the leaf level destructive sampling to quantify photosynthetic pigments was conducted to pick apart the mechanisms contributing to photosynthetic processes of non-photochemical quenching and the resultant changes in observed leaf spectra. At the crown level, Terrestrial Laser Scanning data was used to derive canopy structural information, enabling distance to crown and crown foliage density to be calculated to a fine degree of detail. This information is critical for correcting attenuation of the thermal signal from atmospheric transmission, and to distinguish the relative foliage-to-soil contribution to the thermal and hyperspectral imagery. Ancillary data streams from sap flow and dendrometer devices serve to link leaf, crown and canopy observations.Preliminary results of the leaf and crown level relationships between function and spectra will be discussed. We will demonstrate that operating in a tall canopy (40m) forest can lead to additional complexities. We have found the relationship strength between traditional remote sensing LUE proxies and photosynthetic proxies derived from pigments varies strongly with canopy height and pigment pool size. Additionally, the significance of the relationship between some leaf pigments and spectra hinged upon the inclusion of juvenile or unhealthy leaf samples, which were not representative of the canopy. This has implications for temporal scaling of remote sensing proxies from diurnal to seasonal time frames.

pH dependent unfolding characteristics of DLC8 dimer: Residue level details from NMR.

PubMed

Mohan, P M Krishna; Hosur, Ramakrishna V

2008-11-01

Environment dependence of folding and unfolding of a protein is central to its function. In the same vein, knowledge of pH dependence of stability and folding/unfolding is crucial for many biophysical equilibrium and kinetic studies designed to understand protein folding mechanisms. In the present study we investigated the guanidine induced unfolding transition of dynein light chain protein (DLC8), a cargo adaptor of the dynein complex in the pH range 7-10. It is observed that while the protein remains a dimer in the entire pH range, its stability is somewhat reduced at alkaline pH. Global unfolding features monitored using fluorescence spectroscopy revealed that the unfolding transition of DLC8 at pH 7 is best described by a three-state model, whereas, that at pH 10 is best described by a two-state model. Chemical shift perturbations due to pH change provided insights into the corresponding residue level structural perturbations in the DLC8 dimer. Likewise, backbone (15)N relaxation measurements threw light on the corresponding motional changes in the dimeric protein. These observations have been rationalized on the basis of expected changes with increasing pH in the protonation states of the titratable residues on the structure of the protein. These, in turn provide an explanation for the change from three-state to two-state guanidine induced unfolding transition as the pH is increased from 7 to 10. All these results exemplify and highlight the role of environment vis-à-vis the sequence and structure of a given protein in dictating its folding/unfolding characteristics.

The role of sequence in altering the unfolding pathway of an RNA pseudoknot: a steered molecular dynamics study.

PubMed

Gupta, Asmita; Bansal, Manju

2016-10-19

Mechanical unfolding studies on Ribonucleic Acid (RNA) structures are a subject of tremendous interest as they shed light on the principles of higher order assembly of these structures. Pseudoknotting is one of the most elementary ways in which this higher order assembly is achieved as discrete secondary structural units in RNA are brought in close proximity to form a tertiary structure. Using steered molecular dynamics (SMD) simulations, we have studied the unfolding of five RNA pseudoknot structures that differ from each other either by base substitutions in helices or loops. Our SMD simulations reveal the manner in which a biologically functional RNA pseudoknot unfolds and the effect of changes in the primary structure on this unfolding pathway, providing necessary insights into the driving forces behind the functioning of these structures. We observed that an A → C mutation in the loop sequence makes the pseudoknot far more resistant against force induced disruption relative to its wild type structure. In contrast to this, a base-pair substitution GC → AU near the pseudoknot junction region renders it more vulnerable to this disruption. The quantitative estimation of differences in the unfolding paths was carried out using force extension curves, potential of mean force profiles, and the opening of different Watson-Crick and non-Watson-Crick interactions. The results provide a quantified view in which the unfolding paths of the small RNA structures can be used for investigating the programmability of RNA chains for designing RNA switches and aptamers as their biological folding and unfolding could be assessed and manipulated.

Equilibrium unfolding of A. niger RNase: pH dependence of chemical and thermal denaturation.

PubMed

Kumar, Gundampati Ravi; Sharma, Anurag; Kumari, Moni; Jagannadham, Medicherla V; Debnath, Mira

2011-08-01

Equilibrium unfolding of A. niger RNase with chemical denaturants, for example GuHCl and urea, and thermal unfolding have been studied as a function of pH using fluorescence, far-UV, near-UV, and absorbance spectroscopy. Because of their ability to affect electrostatic interactions, pH and chemical denaturants have a marked effect on the stability, structure, and function of many globular proteins. ANS binding studies have been conducted to enable understanding of the folding mechanism of the protein in the presence of the denaturants. Spectroscopic studies by absorbance, fluorescence, and circular dichroism and use of K2D software revealed that the enzyme has α + β type secondary structure with approximately 29% α-helix, 24% β-sheet, and 47% random coil. Under neutral conditions the enzyme is stable in urea whereas GuHCl-induced equilibrium unfolding was cooperative. A. niger RNase has little ANS binding even under neutral conditions. Multiple intermediates were populated during the pH-induced unfolding of A. niger RNase. Urea and temperature-induced unfolding of A. niger RNase into the molten globule-like state is non-cooperative, in contrast to the cooperativity seen with the native protein, suggesting the presence of two parts/domains, in the molecular structure of A. niger RNase, with different stability that unfolds in steps. Interestingly, the GuHCl-induced unfolding of the A state (molten globule state) of A. niger RNase is unique, because a low concentration of denaturant not only induces structural change but also facilitates transition from one molten globule like state (A(MG1)) into another (I(MG2)).

Gene regulatory network of unfolded protein response genes in endoplasmic reticulum stress.

PubMed

Takayanagi, Sayuri; Fukuda, Riga; Takeuchi, Yuuki; Tsukada, Sakiko; Yoshida, Kenichi

2013-01-01

In the endoplasmic reticulum (ER), secretory and membrane proteins are properly folded and modified, and the failure of these processes leads to ER stress. At the same time, unfolded protein response (UPR) genes are activated to maintain homeostasis. Despite the thorough characterization of the individual gene regulation of UPR genes to date, further investigation of the mutual regulation among UPR genes is required to understand the complex mechanism underlying the ER stress response. In this study, we aimed to reveal a gene regulatory network formed by UPR genes, including immunoglobulin heavy chain-binding protein (BiP), X-box binding protein 1 (XBP1), C/EBP [CCAAT/enhancer-binding protein]-homologous protein (CHOP), PKR-like endoplasmic reticulum kinase (PERK), inositol-requiring 1 (IRE1), activating transcription factor 6 (ATF6), and ATF4. For this purpose, we focused on promoter-luciferase reporters for BiP, XBP1, and CHOP genes, which bear an ER stress response element (ERSE), and p5 × ATF6-GL3, which bears an unfolded protein response element (UPRE). We demonstrated that the luciferase activities of the BiP and CHOP promoters were upregulated by all the UPR genes, whereas those of the XBP1 promoter and p5 × ATF6-GL3 were upregulated by all the UPR genes except for BiP, CHOP, and ATF4 in HeLa cells. Therefore, an ERSE- and UPRE-centered gene regulatory network of UPR genes could be responsible for the robustness of the ER stress response. Finally, we revealed that BiP protein was degraded when cells were treated with DNA-damaging reagents, such as etoposide and doxorubicin; this finding suggests that the expression level of BiP is tightly regulated at the post-translational level, rather than at the transcriptional level, in the presence of DNA damage.

Protein denaturation in vacuo: intrinsic unfolding pathways associated with the native tertiary structure of lysozyme

NASA Astrophysics Data System (ADS)

Arteca, Gustavo A.; Tapia, O.

Using computer-simulated molecular dynamics, we study the effect of sequence mutation on the unfolding mechanism of a native fold. The system considered is the native fold of hen egg-white lysozyme, exposed to centrifugal unfolding in vacuo. This unfolding bias elicits configurational transitions that imitate the behaviour of anhydrous proteins diffusing after electrospraying from neutral-pH solutions. By changing the sequences threaded onto the native fold of lysozyme, we probe the role of disulfide bridges and the effect of a global mutation. We find that the initial denaturing steps share common characteristics for the tested sequences. Recurrent features are: (i) the presence of dumbbell conformers with significant residual secondary structure, (ii) the ubiquitous formation of hairpins and two-stranded β-sheets regardless of disulfide bridges, and (iii) an unfolding pattern where the reduction in folding complexity is highly correlated with the decrease in chain compactness. These findings appear to be intrinsic to the shape of the native fold, suggesting that similar unfolding pathways may be accessible to many protein sequences.

40 CFR 180.422 - Tralomethrin; tolerances for residues.

Code of Federal Regulations, 2013 CFR

2013-07-01

.../revocation date Broccoli 0.5 7/9/13 Cotton, undelinted seed 0.02 7/9/13 Cotton, oil 0.20 7/9/13 Lettuce, head 1.00 7/9/13 Lettuce, leaf 3.00 7/9/13 Soybean, seed 0.05 7/9/13 Sunflower, seed 0.05 7/9/13 (2) A..., manufacturing, and processing establishments, such as restaurants, cafeterias, supermarkets, bakeries, breweries...

40 CFR 180.422 - Tralomethrin; tolerances for residues.

Code of Federal Regulations, 2014 CFR

2014-07-01

.../revocation date Broccoli 0.5 7/9/13 Cotton, undelinted seed 0.02 7/9/13 Cotton, oil 0.20 7/9/13 Lettuce, head 1.00 7/9/13 Lettuce, leaf 3.00 7/9/13 Soybean, seed 0.05 7/9/13 Sunflower, seed 0.05 7/9/13 (2) A..., manufacturing, and processing establishments, such as restaurants, cafeterias, supermarkets, bakeries, breweries...

Effects of Botryosphaeria Inoculation and Stress During Lifting and Outplanting on Initial Growth of Outplanted Sweetgum Seedlings

Treesearch

William Carey; Dave Gerwig; Bert Cregg; Scott Enebak

2004-01-01

A coordinated investigation was carried out with sweetgum seedlings from a nursery in South Carolina (SC) and one in Virginia (VA) to evaluate the effects of cultural practices and Botryosphaeria on the establishment and initial growth of transplanted sweetgum. Seedlings from VA were more sensitive to treatments than those from SC, and date of leaf emergence after...

Multidimensional free energy surface of unfolding of HP-36: Microscopic origin of ruggedness

NASA Astrophysics Data System (ADS)

Ghosh, Rikhia; Roy, Susmita; Bagchi, Biman

2014-10-01

The protein folding funnel paradigm suggests that folding and unfolding proceed as directed diffusion in a multidimensional free energy surface where a multitude of pathways can be traversed during the protein's sojourn from initial to final state. However, finding even a single pathway, with the detail chronicling of intermediates, is an arduous task. In this work we explore the free energy surface of unfolding pathway through umbrella sampling, for a small globular α-helical protein chicken-villin headpiece (HP-36) when the melting of secondary structures is induced by adding DMSO in aqueous solution. We find that the unfolding proceeds through the initial separation or melting of aggregated hydrophobic core that comprises of three phenylalanine residues (Phe7, Phe11, and Phe18). This separation is accompanied by simultaneous melting of the second helix. Unfolding is found to be a multistage process involving crossing of three consecutive minima and two barriers at the initial stage. At a molecular level, Phe18 is observed to reorient itself towards other hydrophobic grooves to stabilize the intermediate states. We identify the configuration of the intermediates and correlate the intermediates with those obtained in our previous works. We also give an estimate of the barriers for different transition states and observe the softening of the barriers with increasing DMSO concentration. We show that higher concentration of DMSO tunes the unfolding pathway by destabilizing the third minimum and stabilizing the second one, indicating the development of a solvent modified, less rugged pathway. The prime outcome of this work is the demonstration that mixed solvents can profoundly transform the nature of the energy landscape and induce unfolding via a modified route. A successful application of Kramer's rate equation correlating the free energy simulation results shows faster rate of unfolding with increasing DMSO concentration. This work perhaps presents the first systematic theoretical study of the effect of a chemical denaturant on the microscopic free energy surface and rates of unfolding of HP-36.

[Flag leaf photosynthetic characteristics, change in chlorophyll fluorescence parameters, and their relationships with yield of winter wheat sowed in spring].

PubMed

Xu, Lan; Gao, Zhi-qang; An, Wei; Li, Yan-liang; Jiao, Xiong-fei; Wang, Chuang-yun

2016-01-01

With five good winter wheat cultivars selected from the middle and lower reaches of Yangtze River and Southwest China as test materials, a field experiment in Xinding basin area of Shanxi Province was conducted to study the photosynthetic characteristics, chlorophyll content, and chlorophyll fluorescence parameters of flag leaf at different sowing dates, as well as the correlations between these indices and yield for two years (2013-2014). The results showed that the difference in most fluorescence parameters except chlorophyll content among cultivars was significant. The correlations between these fluorescence parameters and yield were significant. The variation coefficient of chlorophyll (Chl) content was low (0.12-0.17), and that of performance index based on absorption (PIabs) was high (0.32-0.39), with the partial correlation coefficients of them with grain yield from 2013 to 2014 ranged in 0.70-0.81. Under the early sowing condition, the grain yield positively correlated with PIabs at flowering and filling stages and chlorophyll content at grain filling stage, but negatively correlated with the relative variable fluorescence at I point (Vi) at grain filling stage. About 81.1%-82.8% of grain yield were determined by the variations of PIabs, Chl, and Vi. Wheat cultivars had various performances in the treatments with different sowing dates and a consistent trend was observed in the two experimental years. Among these 5 cultivars, Yangmai 13 was suitable for early sowing, with the flag leaf photosynthetic rate (Pn), Chl, most fluorescence parame-ters, and grain yield showed obviously high levels. In conclusion, under early sowing condition chlorophyll content at grain filling stages, PIabs at flowering and filling stages, and Pn were important indices for selecting wheat cultivars with high photosynthetic efficiency.

Sweepoviruses Cause Disease in Sweet Potato and Related Ipomoea spp.: Fulfilling Koch's Postulates for a Divergent Group in the Genus Begomovirus

PubMed Central

Márquez-Martín, Belén; Moriones, Enrique; Navas-Castillo, Jesús

2011-01-01

Sweet potato (Ipomoea batatas) and related Ipomoea species are frequently infected by monopartite begomoviruses (genus Begomovirus, family Geminiviridae), known as sweepoviruses. Unlike other geminiviruses, the genomes of sweepoviruses have been recalcitrant to rendering infectious clones to date. Thus, Koch's postulates have not been fullfilled for any of the viruses in this group. Three novel species of sweepoviruses have recently been described in Spain: Sweet potato leaf curl Lanzarote virus (SPLCLaV), Sweet potato leaf curl Spain virus (SPLCSV) and Sweet potato leaf curl Canary virus (SPLCCaV). Here we describe the generation of the first infectious clone of an isolate (ES:MAL:BG30:06) of SPLCLaV. The clone consisted of a complete tandem dimeric viral genome in a binary vector. Successful infection by agroinoculation of several species of Ipomoea (including sweet potato) and Nicotiana benthamiana was confirmed by PCR, dot blot and Southern blot hybridization. Symptoms observed in infected plants consisted of leaf curl, yellowing, growth reduction and vein yellowing. Two varieties of sweet potato, ‘Beauregard’ and ‘Promesa’, were infected by agroinoculation, and symptoms of leaf curl and interveinal loss of purple colouration were observed, respectively. The virus present in agroinfected plants was readily transmitted by the whitefly Bemisia tabaci to I. setosa plants. The progeny virus population present in agroinfected I. setosa and sweet potato plants was isolated and identity to the original isolate was confirmed by sequencing. Therefore, Koch's postulates were fulfilled for the first time for a sweepovirus. PMID:22073314

Sweepoviruses cause disease in sweet potato and related Ipomoea spp.: fulfilling Koch's postulates for a divergent group in the genus begomovirus.

PubMed

Trenado, Helena P; Orílio, Anelise F; Márquez-Martín, Belén; Moriones, Enrique; Navas-Castillo, Jesús

2011-01-01

Sweet potato (Ipomoea batatas) and related Ipomoea species are frequently infected by monopartite begomoviruses (genus Begomovirus, family Geminiviridae), known as sweepoviruses. Unlike other geminiviruses, the genomes of sweepoviruses have been recalcitrant to rendering infectious clones to date. Thus, Koch's postulates have not been fullfilled for any of the viruses in this group. Three novel species of sweepoviruses have recently been described in Spain: Sweet potato leaf curl Lanzarote virus (SPLCLaV), Sweet potato leaf curl Spain virus (SPLCSV) and Sweet potato leaf curl Canary virus (SPLCCaV). Here we describe the generation of the first infectious clone of an isolate (ES:MAL:BG30:06) of SPLCLaV. The clone consisted of a complete tandem dimeric viral genome in a binary vector. Successful infection by agroinoculation of several species of Ipomoea (including sweet potato) and Nicotiana benthamiana was confirmed by PCR, dot blot and Southern blot hybridization. Symptoms observed in infected plants consisted of leaf curl, yellowing, growth reduction and vein yellowing. Two varieties of sweet potato, 'Beauregard' and 'Promesa', were infected by agroinoculation, and symptoms of leaf curl and interveinal loss of purple colouration were observed, respectively. The virus present in agroinfected plants was readily transmitted by the whitefly Bemisia tabaci to I. setosa plants. The progeny virus population present in agroinfected I. setosa and sweet potato plants was isolated and identity to the original isolate was confirmed by sequencing. Therefore, Koch's postulates were fulfilled for the first time for a sweepovirus.

Mechanical unfolding reveals stable 3-helix intermediates in talin and α-catenin

PubMed Central

2018-01-01

Mechanical stability is a key feature in the regulation of structural scaffolding proteins and their functions. Despite the abundance of α-helical structures among the human proteome and their undisputed importance in health and disease, the fundamental principles of their behavior under mechanical load are poorly understood. Talin and α-catenin are two key molecules in focal adhesions and adherens junctions, respectively. In this study, we used a combination of atomistic steered molecular dynamics (SMD) simulations, polyprotein engineering, and single-molecule atomic force microscopy (smAFM) to investigate unfolding of these proteins. SMD simulations revealed that talin rod α-helix bundles as well as α-catenin α-helix domains unfold through stable 3-helix intermediates. While the 5-helix bundles were found to be mechanically stable, a second stable conformation corresponding to the 3-helix state was revealed. Mechanically weaker 4-helix bundles easily unfolded into a stable 3-helix conformation. The results of smAFM experiments were in agreement with the findings of the computational simulations. The disulfide clamp mutants, designed to protect the stable state, support the 3-helix intermediate model in both experimental and computational setups. As a result, multiple discrete unfolding intermediate states in the talin and α-catenin unfolding pathway were discovered. Better understanding of the mechanical unfolding mechanism of α-helix proteins is a key step towards comprehensive models describing the mechanoregulation of proteins. PMID:29698481

Frost sensitivity of various deciduous plant species during leaf development in spring

NASA Astrophysics Data System (ADS)

Estrella, Nicole; Heinzmann, Verena; Menzel, Annette

2017-04-01

Frost damage in deciduous woody plants is a major climate component affecting fitness and distribution of species. It is a trade-off between early bud burst enlarging the potential growing season and frost risk for deciduous plants in many regions. In a warming world observed earlier budburst may lead to an increased risk of spring frost damage caused by higher variability in temperatures (IPCC 2007). Lenz et al. (2013) showed that leaves are in general more sensitive to frost in later leaf development stages. But still there is little knowledge on stages of leaf development and their susceptibility to frost damage in many deciduous species. Additionally there might be variation with plant traits or different strategies within specific groups of species. Frost risk minimization can also be achieved by variability in bud burst within a specimen. Therefore, in this study we observed more than 174 individual plant specimen of 96 deciduous woody plant species growing in a comparable microclimate outside on the campus of the Technical University of Munich in Freising, southern Germany. Their phenology was intensively studied from 12th of March to 4th of May, including variation within a specimen. Several times twigs for the frost experiment were cut in different stages of leaf development and exposed to freezing temperatures of -4 and -6°C in two lab freezers. Since the leaf development in spring 2015 started comparably late, too many species emerged simultaneously leading to some capacity problems in the freezers. Nevertheless, our results still reveal novel aspects concerning leaf development and frost sensitivity. The phenological development proceeded in general from outside to inside of the crown (59%), in 33% of the cases all over the plant simultaneously. Sporadic, inside to outside or vertical development characteristics occurred in rare cases (8%). Mixed model analysis indicated impacts on phenology by plant family, natural origin, pollination mode, and development characteristic (in decreasing order of significance). The frost experiment clearly showed that damage at -6°C was larger than at -4°C and that twigs frosted at later dates, thus in higher phenological development stages, were more prone to frost damage than twigs frosted at earlier dates / in lower development stages. Additionally, there was a phylogenetic effect since frost damage significantly varied with plant family whereas plant origin had no relevance. References Lenz A, Hoch G, Vitasse Y, Körner C (2013) European deciduous trees exhibit similar safety margins against damage by spring freeze events along elevational gradients. New Phytologist 200: 1166-1175. Stocker T, Qin D, Platner G (2013) Climate change 2013: The physical science basis. Working Group I Contribution to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Summary for Policymakers (IPCC, 2013).

The Graded Unfolding Model: A Unidimensional Item Response Model for Unfolding Graded Responses.

ERIC Educational Resources Information Center

Roberts, James S.; Laughlin, James E.

Binary or graded disagree-agree responses to attitude items are often collected for the purpose of attitude measurement. Although such data are sometimes analyzed with cumulative measurement models, recent investigations suggest that unfolding models are more appropriate (J. S. Roberts, 1995; W. H. Van Schuur and H. A. L. Kiers, 1994). Advances in…

A Unidimensional Item Response Model for Unfolding Responses from a Graded Disagree-Agree Response Scale.

ERIC Educational Resources Information Center

Roberts, James S.; Laughlin, James E.

1996-01-01

A parametric item response theory model for unfolding binary or graded responses is developed. The graded unfolding model (GUM) is a generalization of the hyperbolic cosine model for binary data of D. Andrich and G. Luo (1993). Applicability of the GUM to attitude testing is illustrated with real data. (SLD)

Immobilized unfolded cytochrome c acts as a catalyst for dioxygen reduction.

PubMed

Tavagnacco, Claudio; Monari, Stefano; Ranieri, Antonio; Bortolotti, Carlo Augusto; Peressini, Silvia; Borsari, Marco

2011-10-21

Unfolding turns immobilized cytochrome c into a His-His ligated form endowed with catalytic activity towards O(2), which is absent in the native protein. Dioxygen could be used by naturally occurring unfolded cytochrome c as a substrate for the production of partially reduced oxygen species (PROS) contributing to the cell oxidative stress.

Using Data Augmentation and Markov Chain Monte Carlo for the Estimation of Unfolding Response Models

ERIC Educational Resources Information Center

Johnson, Matthew S.; Junker, Brian W.

2003-01-01

Unfolding response models, a class of item response theory (IRT) models that assume a unimodal item response function (IRF), are often used for the measurement of attitudes. Verhelst and Verstralen (1993)and Andrich and Luo (1993) independently developed unfolding response models by relating the observed responses to a more common monotone IRT…

Multidimensional Unfolding by Nonmetric Multidimensional Scaling of Spearman Distances in the Extended Permutation Polytope

ERIC Educational Resources Information Center

Van Deun, Katrijn; Heiser, Willem J.; Delbeke, Luc

2007-01-01

A multidimensional unfolding technique that is not prone to degenerate solutions and is based on multidimensional scaling of a complete data matrix is proposed: distance information about the unfolding data and about the distances both among judges and among objects is included in the complete matrix. The latter information is derived from the…

Modified Likelihood-Based Item Fit Statistics for the Generalized Graded Unfolding Model

ERIC Educational Resources Information Center

Roberts, James S.

2008-01-01

Orlando and Thissen (2000) developed an item fit statistic for binary item response theory (IRT) models known as S-X[superscript 2]. This article generalizes their statistic to polytomous unfolding models. Four alternative formulations of S-X[superscript 2] are developed for the generalized graded unfolding model (GGUM). The GGUM is a…

The Random-Threshold Generalized Unfolding Model and Its Application of Computerized Adaptive Testing

ERIC Educational Resources Information Center

Wang, Wen-Chung; Liu, Chen-Wei; Wu, Shiu-Lien

2013-01-01

The random-threshold generalized unfolding model (RTGUM) was developed by treating the thresholds in the generalized unfolding model as random effects rather than fixed effects to account for the subjective nature of the selection of categories in Likert items. The parameters of the new model can be estimated with the JAGS (Just Another Gibbs…

Unfolding linac photon spectra and incident electron energies from experimental transmission data, with direct independent validation

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

Ali, E. S. M.; McEwen, M. R.; Rogers, D. W. O.

2012-11-15

Purpose: In a recent computational study, an improved physics-based approach was proposed for unfolding linac photon spectra and incident electron energies from transmission data. In this approach, energy differentiation is improved by simultaneously using transmission data for multiple attenuators and detectors, and the unfolding robustness is improved by using a four-parameter functional form to describe the photon spectrum. The purpose of the current study is to validate this approach experimentally, and to demonstrate its application on a typical clinical linac. Methods: The validation makes use of the recent transmission measurements performed on the Vickers research linac of National Research Councilmore » Canada. For this linac, the photon spectra were previously measured using a NaI detector, and the incident electron parameters are independently known. The transmission data are for eight beams in the range 10-30 MV using thick Be, Al and Pb bremsstrahlung targets. To demonstrate the approach on a typical clinical linac, new measurements are performed on an Elekta Precise linac for 6, 10 and 25 MV beams. The different experimental setups are modeled using EGSnrc, with the newly added photonuclear attenuation included. Results: For the validation on the research linac, the 95% confidence bounds of the unfolded spectra fall within the noise of the NaI data. The unfolded spectra agree with the EGSnrc spectra (calculated using independently known electron parameters) with RMS energy fluence deviations of 4.5%. The accuracy of unfolding the incident electron energy is shown to be {approx}3%. A transmission cutoff of only 10% is suitable for accurate unfolding, provided that the other components of the proposed approach are implemented. For the demonstration on a clinical linac, the unfolded incident electron energies and their 68% confidence bounds for the 6, 10 and 25 MV beams are 6.1 {+-} 0.1, 9.3 {+-} 0.1, and 19.3 {+-} 0.2 MeV, respectively. The unfolded spectra for the clinical linac agree with the EGSnrc spectra (calculated using the unfolded electron energies) with RMS energy fluence deviations of 3.7%. The corresponding measured and EGSnrc-calculated transmission data agree within 1.5%, where the typical transmission measurement uncertainty on the clinical linac is 0.4% (not including the uncertainties on the incident electron parameters). Conclusions: The approach proposed in an earlier study for unfolding photon spectra and incident electron energies from transmission data is accurate and practical for clinical use.« less

Forced-Unfolding and Force-Quench Refolding of RNA Hairpins

PubMed Central

Hyeon, Changbong; Thirumalai, D.

2006-01-01

Nanomanipulation of individual RNA molecules, using laser optical tweezers, has made it possible to infer the major features of their energy landscape. Time-dependent mechanical unfolding trajectories, measured at a constant stretching force (fS) of simple RNA structures (hairpins and three-helix junctions) sandwiched between RNA/DNA hybrid handles show that they unfold in a reversible all-or-none manner. To provide a molecular interpretation of the experiments we use a general coarse-grained off-lattice Gō-like model, in which each nucleotide is represented using three interaction sites. Using the coarse-grained model we have explored forced-unfolding of RNA hairpin as a function of fS and the loading rate (rf). The simulations and theoretical analysis have been done both with and without the handles that are explicitly modeled by semiflexible polymer chains. The mechanisms and timescales for denaturation by temperature jump and mechanical unfolding are vastly different. The directed perturbation of the native state by fS results in a sequential unfolding of the hairpin starting from their ends, whereas thermal denaturation occurs stochastically. From the dependence of the unfolding rates on rf and fS we show that the position of the unfolding transition state is not a constant but moves dramatically as either rf or fS is changed. The transition-state movements are interpreted by adopting the Hammond postulate for forced-unfolding. Forced-unfolding simulations of RNA, with handles attached to the two ends, show that the value of the unfolding force increases (especially at high pulling speeds) as the length of the handles increases. The pathways for refolding of RNA from stretched initial conformation, upon quenching fS to the quench force fQ, are highly heterogeneous. The refolding times, upon force-quench, are at least an order-of-magnitude greater than those obtained by temperature-quench. The long fQ-dependent refolding times starting from fully stretched states are analyzed using a model that accounts for the microscopic steps in the rate-limiting step, which involves the trans to gauche transitions of the dihedral angles in the GAAA tetraloop. The simulations with explicit molecular model for the handles show that the dynamics of force-quench refolding is strongly dependent on the interplay of their contour length and persistence length and the RNA persistence length. Using the generality of our results, we also make a number of precise experimentally testable predictions. PMID:16473903

Unfolding single RNA molecules: bridging the gap between equilibrium and non-equilibrium statistical thermodynamics.

PubMed

Bustamante, Carlos

2005-11-01

During the last 15 years, scientists have developed methods that permit the direct mechanical manipulation of individual molecules. Using this approach, they have begun to investigate the effect of force and torque in chemical and biochemical reactions. These studies span from the study of the mechanical properties of macromolecules, to the characterization of molecular motors, to the mechanical unfolding of individual proteins and RNA. Here I present a review of some of our most recent results using mechanical force to unfold individual molecules of RNA. These studies make it possible to follow in real time the trajectory of each molecule as it unfolds and characterize the various intermediates of the reaction. Moreover, if the process takes place reversibly it is possible to extract both kinetic and thermodynamic information from these experiments at the same time that we characterize the forces that maintain the three-dimensional structure of the molecule in solution. These studies bring us closer to the biological unfolding processes in the cell as they simulate in vitro, the mechanical unfolding of RNAs carried out in the cell by helicases. If the unfolding process occurs irreversibly, I show here that single-molecule experiments can still provide equilibrium, thermodynamic information from non-equilibrium data by using recently discovered fluctuation theorems. Such theorems represent a bridge between equilibrium and non-equilibrium statistical mechanics. In fact, first derived in 1997, the first experimental demonstration of the validity of fluctuation theorems was obtained by unfolding mechanically a single molecule of RNA. It is perhaps a sign of the times that important physical results are these days used to extract information about biological systems and that biological systems are being used to test and confirm fundamental new laws in physics.

Investigating the structural transitions of proteins during dissolution by mass spectrometry.

PubMed

Gong, Xiaoyun; Xiong, Xingchuang; Qi, Lin; Fang, Xiang

2017-03-01

An appropriate solvent environment is essential for the implementation of biological functions of proteins. Interactions between protein residues and solvent molecules are of great importance for proteins to maintain their active structure and catalyze biochemical reactions. In this study, we investigated such interactions and studied the structural transitions of proteins during their dissolution process. Our previously developed technique, namely solvent assisted electric field induced desorption/ionization, was used for the dissolution and immediate ionization of proteins. Different solvents and proteins were involved in the investigation. According to the results, cytochrome c underwent significant unfolding during dissolution in the most commonly used NH 4 Ac buffer. The unfolding got more serious when the concentration of NH 4 Ac was further increased. Extending the dissolution time resulted in the re-folding of cytochrome c. In comparison, no unfolding was observed if cytochrome c was pre-dissolved in NH 4 Ac buffer and detected by nano-ESI. Furthermore, no unfolding was observed during the dissolution process of cytochrome c in water. Interactions between the residues of cytochrome c and the solute of NH 4 Ac might be the reason for the unfolding phenomenon. Similar unfolding phenomenon was observed on holo-myoglobin. However, the observed dissolution feature of insulin was different. No unfolding was observed on insulin during dissolution in NH 4 Ac buffers. Insulin underwent observable unfolding when water was used for dissolution. This might be due to the structural difference between different proteins. The obtained results in the present study furthered our insights into the interactions between proteins and the solvents during the phase transition of dissolution. Copyright © 2016 Elsevier B.V. All rights reserved.

High stability of a ferredoxin from the hyperthermophilic archaeon A. ambivalens: Involvement of electrostatic interactions and cofactors

PubMed Central

Moczygemba, Charmaine; Guidry, Jesse; Jones, Kathryn L.; Gomes, Cláudio M.; Teixeira, Miguel; Wittung-Stafshede, Pernilla

2001-01-01

The ferredoxin from the thermophilic archaeon Acidianus ambivalens is a small monomeric seven-iron protein with a thermal midpoint (Tm) of 122°C (pH 7). To gain insight into the basis of its thermostability, we have characterized unfolding reactions induced chemically and thermally at various pHs. Thermal unfolding of this ferredoxin, in the presence of various guanidine hydrochloride (GuHCl) concentrations, yields a linear correlation between unfolding enthalpies (ΔH[Tm]) and Tm from which an upper limit for the heat capacity of unfolding (ΔCP) was determined to be 3.15 ± 0.1 kJ/(mole • K). Only by the use of the stronger denaturant guanidine thiocyanate (GuSCN) is unfolding of A. ambivalens ferredoxin at pH 7 (20°C) observed ([GuSCN]1/2 = 3.1 M; ΔGU[H2O] = 79 ± 8 kJ/mole). The protein is, however, less stable at low pH: At pH 2.5, Tm is 64 ± 1°C, and GuHCl-induced unfolding shows a midpoint at 2.3 M (ΔGU[H2O] = 20 ± 1 kJ/mole). These results support that electrostatic interactions contribute significantly to the stability. Analysis of the three-dimensional molecular model of the protein shows that there are several possible ion pairs on the surface. In addition, ferredoxin incorporates two iron–sulfur clusters and a zinc ion that all coordinate deprotonated side chains. The zinc remains bound in the unfolded state whereas the iron–sulfur clusters transiently form linear three-iron species (in pH range 2.5 to 10), which are associated with the unfolded polypeptide, before their complete degradation. PMID:11468351

Microscopic dynamics of water around unfolded structures of barstar at room temperature

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

Pal, Somedatta; Chakraborty, Kaushik; Khatua, Prabir

2015-02-07

The breaking of the native structure of a protein and its influences on the dynamic response of the surrounding solvent is an important issue in protein folding. In this work, we have carried out atomistic molecular dynamics simulations to unfold the protein barstar at two different temperatures (400 K and 450 K). The two unfolded forms obtained at such high temperatures are further studied at room temperature to explore the effects of nonuniform unfolding of the protein secondary structures along two different pathways on the microscopic dynamical properties of the surface water molecules. It is demonstrated that though the structuralmore » transition of the protein in general results in less restricted water motions around its segments, but there are evidences of formation of new conformational motifs upon unfolding with increasingly confined environment around them, thereby resulting in further restricted water mobility in their hydration layers. Moreover, it is noticed that the effects of nonuniform unfolding of the protein segments on the relaxation times of the protein–water (PW) and the water–water (WW) hydrogen bonds are correlated with hindered hydration water motions. However, the kinetics of breaking and reformation of such hydrogen bonds are found to be influenced differently at the interface. It is observed that while the effects of unfolding on the PW hydrogen bond kinetics seem to be minimum, but the kinetics involving the WW hydrogen bonds around the protein segments exhibit noticeably heterogeneous characteristics. We believe that this is an important observation, which can provide valuable insights on the origin of heterogeneous influence of unfolding of a protein on the microscopic properties of its hydration water.« less

Unfolding single- and multilayers

NASA Astrophysics Data System (ADS)

Llorens, Maria-Gema; Bons, Paul D.; Griera, Albert; Gomez-Rivas, Enrique

2014-05-01

When planar structures (e.g. sedimentary layers, veins, dykes, cleavages, etc.) are subjected to deformation, they have about equal chances to be shortened or stretched. The most common shortening and stretching structures are folds and boudinage, respectively. However, boudinage requires additional deformation mechanisms apart from viscous flow, like formation of fractures or strain localization. When folded layers are subjected to extension, they could potentially unfold back to straight layers. Although probably not uncommon, this would be difficult to recognize. Open questions are whether folded layers can unfold, what determines their mechanical behaviour and how we can recognize them in the field. In order to approach these questions, we present a series of numerical experiments that simulate stretching of previously folded single- and multi-layers in simple shear, using the two dimensional numerical modelling platform ELLE, including the finite element module BASIL that calculates viscous deformation. We investigate the parameters that affect a fold train once it rotates into the extensional field. The results show that the unfolding process strongly depends on the viscosity contrast between the layer and matrix (Llorens et al., 2013). Layers do not completely unfold when they experience softening before or during the stretching process or when other neighbouring competent layers prevent them from unfolding. The foliation refraction patterns are the main indicators of unfolded folds. Additionally, intrafolial folds and cusp-like folds adjacent to straight layers, as well as variations in fold amplitudes and limb lengths of irregular folds can also be used as indicators of stretching of a layer after shortening and folding. References: Llorens, M-.G., Bons, P.D., Griera, A. and Gomez-Rivas, E. 2013. When do folds unfold during progressive shear?. Geology, 41, 563-566.

The fluorescence intensities ratio is not a reliable parameter for evaluation of protein unfolding transitions.

PubMed

Žoldák, Gabriel; Jancura, Daniel; Sedlák, Erik

2017-06-01

Monitoring the fluorescence of proteins, particularly the fluorescence of intrinsic tryptophan residues, is a popular method often used in the analysis of unfolding transitions (induced by temperature, chemical denaturant, and pH) in proteins. The tryptophan fluorescence provides several suitable parameters, such as steady-state fluorescence intensity, apparent quantum yield, mean fluorescence lifetime, position of emission maximum that are often utilized for the observation of the conformational/unfolding transitions of proteins. In addition, the fluorescence intensities ratio at different wavelengths (usually at 330 nm and 350 nm) is becoming an increasingly popular parameter for the evaluation of thermal transitions. We show that, under certain conditions, the use of this parameter for the analysis of unfolding transitions leads to the incorrect determination of thermodynamic parameters characterizing unfolding transitions in proteins (e.g., melting temperature) and, hence, can compromise the hit identification during high-throughput drug screening campaigns. © 2017 The Protein Society.

Measurement of 241Am-Be spectra (bare and Pb-covered) using TLD pairs in multi-spheres: Spectrum unfolding by different methods

NASA Astrophysics Data System (ADS)

Tripathy, S. P.; Bakshi, A. K.; Sathian, V.; Tripathi, S. M.; Vega-carrillo, H. R.; Nandy, M.; Sarkar, P. K.; Sharma, D. N.

2009-01-01

The neutron spectra from a Pb-covered and a bare (without Pb-cover) 241Am-Be (α,n) source were measured using thermoluminescent detector (TLD) pairs of 6LiF and 7LiF with high-density polyethylene (HDPE) multi-spheres of seven different diameters. A total of 8 distinct neutron response signals (including a bare mode exposure) were obtained from which the energy distribution for the entire energy range was generated with the help of different neutron spectrum unfolding methods, viz. BUNKI, BUNKIUT and Frascati unfolding interactive tool (FRUIT). Shape of these spectra are matching very well and is also comparable with the standard IAEA 241Am-Be spectrum, thus, validating the unfolding methods used in this work. The effect of Pb-cover on the spectrum and the unfolding details are reported in the paper.

GroEL stimulates protein folding through forced unfolding

PubMed Central

Lin, Zong; Madan, Damian; Rye, Hays S

2013-01-01

Many proteins cannot fold without the assistance of chaperonin machines like GroEL and GroES. The nature of this assistance, however, remains poorly understood. Here we demonstrate that unfolding of a substrate protein by GroEL enhances protein folding. We first show that capture of a protein on the open ring of a GroEL–ADP–GroES complex, GroEL’s physiological acceptor state for non-native proteins in vivo, leaves the substrate protein in an unexpectedly compact state. Subsequent binding of ATP to the same GroEL ring causes rapid, forced unfolding of the substrate protein. Notably, the fraction of the substrate protein that commits to the native state following GroES binding and protein release into the GroEL–GroES cavity is proportional to the extent of substrate-protein unfolding. Forced protein unfolding is thus a central component of the multilayered stimulatory mechanism used by GroEL to drive protein folding. PMID:18311152

Toward an atomistic description of the urea-denatured state of proteins.

PubMed

Candotti, Michela; Esteban-Martín, Santiago; Salvatella, Xavier; Orozco, Modesto

2013-04-09

We present here the characterization of the structural, dynamics, and energetics of properties of the urea-denatured state of ubiquitin, a small prototypical soluble protein. By combining state-of-the-art molecular dynamics simulations with NMR and small-angle X-ray scattering data, we were able to: (i) define the unfolded state ensemble, (ii) understand the energetics stabilizing unfolded structures in urea, (iii) describe the dedifferential nature of the interactions of the fully unfolded proteins with urea and water, and (iv) characterize the early stages of protein refolding when chemically denatured proteins are transferred to native conditions. The results presented herein are unique in providing a complete picture of the chemically unfolded state of proteins and contribute to deciphering the mechanisms that stabilize the native state of proteins, as well as those that maintain them unfolded in the presence of urea.

Toward an atomistic description of the urea-denatured state of proteins

PubMed Central

Candotti, Michela; Esteban-Martín, Santiago; Salvatella, Xavier; Orozco, Modesto

2013-01-01

We present here the characterization of the structural, dynamics, and energetics of properties of the urea-denatured state of ubiquitin, a small prototypical soluble protein. By combining state-of-the-art molecular dynamics simulations with NMR and small-angle X-ray scattering data, we were able to: (i) define the unfolded state ensemble, (ii) understand the energetics stabilizing unfolded structures in urea, (iii) describe the dedifferential nature of the interactions of the fully unfolded proteins with urea and water, and (iv) characterize the early stages of protein refolding when chemically denatured proteins are transferred to native conditions. The results presented herein are unique in providing a complete picture of the chemically unfolded state of proteins and contribute to deciphering the mechanisms that stabilize the native state of proteins, as well as those that maintain them unfolded in the presence of urea. PMID:23536295

Structural basis for the unfolding of anthrax lethal factor by protective antigen oligomers

PubMed Central

Feld, Geoffrey K.; Thoren, Katie L.; Kintzer, Alexander F.; Sterling, Harry J.; Tang, Iok I.; Greenberg, Shoshana G.; Williams, Evan R.; Krantz, Bryan A.

2011-01-01

The protein transporter, anthrax lethal toxin, is comprised of protective antigen (PA), a transmembrane translocase, and lethal factor (LF), a cytotoxic enzyme. Following assembly into holotoxin complexes, PA forms an oligomeric channel that unfolds LF and translocates it into the host cell. We report the crystal structure of the core of a lethal toxin complex to 3.1-Å resolution; the structure contains a PA octamer bound to four LF PA-binding domains (LFN). The first α helix and β strand of each LFN unfold and dock into a deep amphipathic cleft on the surface of the PA octamer, which we call the α clamp. The α clamp possesses nonspecific polypeptide binding activity and is functionally relevant to efficient holotoxin assembly, PA octamer formation, and LF unfolding and translocation. This structure provides insight on the mechanism of translocation-coupled protein unfolding. PMID:21037566

Signatures of unfolding in the early stages of protein denaturation

NASA Astrophysics Data System (ADS)

Gray, Harry B.; Winkler, Jay R.; Kozak, John J.

2012-04-01

A comparative study of the early stages of unfolding of five proteins: cyt c, c-b 562, cyt c‧, azurin, and lysozyme is reported. From crystallographic data, helical regions and intervening non-helical (or 'turning') regions are identified in each. Exploiting a previously introduced geometrical model, the paper describes quantitatively the stepwise extension of a polypeptide chain subject to the geometrical constraint that the spatial relationship among the residues of each triplet is fixed by native-state crystallographic data. Despite differences among the above-cited proteins, remarkable universality of behavior is found in the early stages of unfolding. At the very earliest stages, internal residues in each helical region have a common unfolding history; the terminal residues, however, are extraordinarily sensitive to structural perturbations. Residues in non-helical sections of the polypeptide unfold after residues in the internal helical regions, but with increasing steric perturbation playing a dominant role in advancing denaturation.

Development and Application of a High Throughput Protein Unfolding Kinetic Assay

PubMed Central

Wang, Qiang; Waterhouse, Nicklas; Feyijinmi, Olusegun; Dominguez, Matthew J.; Martinez, Lisa M.; Sharp, Zoey; Service, Rachel; Bothe, Jameson R.; Stollar, Elliott J.

2016-01-01

The kinetics of folding and unfolding underlie protein stability and quantification of these rates provides important insights into the folding process. Here, we present a simple high throughput protein unfolding kinetic assay using a plate reader that is applicable to the studies of the majority of 2-state folding proteins. We validate the assay by measuring kinetic unfolding data for the SH3 (Src Homology 3) domain from Actin Binding Protein 1 (AbpSH3) and its stabilized mutants. The results of our approach are in excellent agreement with published values. We further combine our kinetic assay with a plate reader equilibrium assay, to obtain indirect estimates of folding rates and use these approaches to characterize an AbpSH3-peptide hybrid. Our high throughput protein unfolding kinetic assays allow accurate screening of libraries of mutants by providing both kinetic and equilibrium measurements and provide a means for in-depth ϕ-value analyses. PMID:26745729

Expansion and internal friction in unfolded protein chain.

PubMed

Yasin, U Mahammad; Sashi, Pulikallu; Bhuyan, Abani K

2013-10-10

Similarities in global properties of homopolymers and unfolded proteins provide approaches to mechanistic description of protein folding. Here, hydrodynamic properties and relaxation rates of the unfolded state of carbonmonoxide-liganded cytochrome c (cyt-CO) have been measured using nuclear magnetic resonance and laser photolysis methods. Hydrodynamic radius of the unfolded chain gradually increases as the solvent turns increasingly better, consistent with theory. Curiously, however, the rate of intrachain contact formation also increases with an increasing denaturant concentration, which, by Szabo, Schulten, and Schulten theory for the rate of intramolecular contact formation in a Gaussian polymer, indicates growing intramolecular diffusion. It is argued that diminishing nonbonded atom interactions with increasing denaturant reduces internal friction and, thus, increases the rate of polypeptide relaxation. Qualitative scaling of the extent of unfolding with nonbonded repulsions allows for description of internal friction by a phenomenological model. The degree of nonbonded atom interactions largely determines the extent of internal friction.

Soft Vibrational Modes Predict Breaking Events during Force-Induced Protein Unfolding.

PubMed

Habibi, Mona; Plotkin, Steven S; Rottler, Jörg

2018-02-06

We investigate the correlation between soft vibrational modes and unfolding events in simulated force spectroscopy of proteins. Unfolding trajectories are obtained from molecular dynamics simulations of a Gō model of a monomer of a mutant of superoxide dismutase 1 protein containing all heavy atoms in the protein, and a normal mode analysis is performed based on the anisotropic network model. We show that a softness map constructed from the superposition of the amplitudes of localized soft modes correlates with unfolding events at different stages of the unfolding process. Soft residues are up to eight times more likely to undergo disruption of native structure than the average amino acid. The memory of the softness map is retained for extensions of up to several nanometers, but decorrelates more rapidly during force drops. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Climatic Implications of Macro- and Microfossil Assemblages from Late Pleistocene Deposits in Northern New Zealand

NASA Astrophysics Data System (ADS)

Ogden, John; Newnham, Rewi M.; Palmer, Jonathan G.; Serra, Richard G.; Mitchell, Neil D.

1993-01-01

Twenty-two plant species were identified from leaves, fruits, or flowers, and 41 taxa from pollen, present in a macrofossil (leaf) layer in a peat swamp formed on Pleistocene dunes on the Aupouri Peninsula in northern New Zealand. Eight genera of gymnosperms are represented. With the exception of Lagarostrobos colensoi, all tree species abundant as macrofossils are also common as pollen. Macrofossils enabled the on-site flora to be compared with the regional flora, represented by the pollen rain. Studies on leaf decomposition rates indicate bias toward sclerophyllous species in the macrofossils. Identification to species level and treering data from preserved kauri logs allow quantitative comparisons with similar extant communities. Current climatic conditions at those analogue sites are cooler (2° to 3°C), cloudier (11%), and much wetter (85%) than those currently prevailing on the Aupouri Peninsula. Dendrochronological results also suggest that the far north of New Zealand had a cooler, cloudier, and wetter climate at the time the fossil leaf assemblage was formed. Radiocarbon dates from possibly contaminated samples suggest that a diverse mixed gymnosperm/angiosperm forest, dominated by kauri ( Agathis australis), was present about (or sometime before) 41,00034,000 yr B.P., when the leaf layer was formed. Similar temperature reductions have been postulated for this period in New Zealand by other authors.

Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches*

PubMed Central

Ben Chobba, Ines; Elleuch, Amine; Ayadi, Imen; Khannous, Lamia; Namsi, Ahmed; Cerqueira, Frederique; Drira, Noureddine; Gharsallah, Néji; Vallaeys, Tatiana

2013-01-01

Endophytic flora plays a vital role in the colonization and survival of host plants, especially in harsh environments, such as arid regions. This flora may, however, contain pathogenic species responsible for various troublesome host diseases. The present study is aimed at investigating the diversity of both cultivable and non-cultivable endophytic fungal floras in the internal tissues (roots and leaves) of Tunisian date palm trees (Phoenix dactylifera). Accordingly, 13 isolates from both root and leaf samples, exhibiting distinct colony morphology, were selected from potato dextrose agar (PDA) medium and identified by a sequence match search wherein their 18S–28S internal transcribed spacer (ITS) sequences were compared to those available in public databases. These findings revealed that the cultivable root and leaf isolates fell into two groups, namely Nectriaceae and Pleosporaceae. Additionally, total DNA from palm roots and leaves was further extracted and ITS fragments were amplified. Restriction fragment length polymorphism (RFLP) analysis of the ITS from 200 fungal clones (leaves: 100; roots: 100) using HaeIII restriction enzyme revealed 13 distinct patterns that were further sequenced and led to the identification of Alternaria, Cladosporium, Davidiella (Cladosporium teleomorph), Pythium, Curvularia, and uncharacterized fungal endophytes. Both approaches confirmed that while the roots were predominantly colonized by Fusaria (members of the Nectriaceae family), the leaves were essentially colonized by Alternaria (members of the Pleosporaceae family). Overall, the findings of the present study constitute, to the authors’ knowledge, the first extensive report on the diversity of endophytic fungal flora associated with date palm trees (P. dactylifera). PMID:24302709

Review of MEMS differential scanning calorimetry for biomolecular study

NASA Astrophysics Data System (ADS)

Yu, Shifeng; Wang, Shuyu; Lu, Ming; Zuo, Lei

2017-12-01

Differential scanning calorimetry (DSC) is one of the few techniques that allow direct determination of enthalpy values for binding reactions and conformational transitions in biomolecules. It provides the thermodynamics information of the biomolecules which consists of Gibbs free energy, enthalpy and entropy in a straightforward manner that enables deep understanding of the structure function relationship in biomolecules such as the folding/unfolding of protein and DNA, and ligand bindings. This review provides an up to date overview of the applications of DSC in biomolecular study such as the bovine serum albumin denaturation study, the relationship between the melting point of lysozyme and the scanning rate. We also introduce the recent advances of the development of micro-electro-mechanic-system (MEMS) based DSCs.

Spectral analysis of amazon canopy phenology during the dry season using a tower hyperspectral camera and modis observations

NASA Astrophysics Data System (ADS)

de Moura, Yhasmin Mendes; Galvão, Lênio Soares; Hilker, Thomas; Wu, Jin; Saleska, Scott; do Amaral, Cibele Hummel; Nelson, Bruce Walker; Lopes, Aline Pontes; Wiedeman, Kenia K.; Prohaska, Neill; de Oliveira, Raimundo Cosme; Machado, Carolyne Bueno; Aragão, Luiz E. O. C.

2017-09-01

The association between spectral reflectance and canopy processes remains challenging for quantifying large-scale canopy phenological cycles in tropical forests. In this study, we used a tower-mounted hyperspectral camera in an eastern Amazon forest to assess how canopy spectral signals of three species are linked with phenological processes in the 2012 dry season. We explored different approaches to disentangle the spectral components of canopy phenology processes and analyze their variations over time using 17 images acquired by the camera. The methods included linear spectral mixture analysis (SMA); principal component analysis (PCA); continuum removal (CR); and first-order derivative analysis. In addition, three vegetation indices potentially sensitive to leaf flushing, leaf loss and leaf area index (LAI) were calculated: the Enhanced Vegetation Index (EVI), Normalized Difference Vegetation Index (NDVI) and the entitled Green-Red Normalized Difference (GRND) index. We inspected also the consistency of the camera observations using Moderate Resolution Imaging Spectroradiometer (MODIS) and available phenological data on new leaf production and LAI of young, mature and old leaves simulated by a leaf demography-ontogeny model. The results showed a diversity of phenological responses during the 2012 dry season with related changes in canopy structure and greenness values. Because of the differences in timing and intensity of leaf flushing and leaf shedding, Erisma uncinatum, Manilkara huberi and Chamaecrista xinguensis presented different green vegetation (GV) and non-photosynthetic vegetation (NPV) SMA fractions; distinct PCA scores; changes in depth, width and area of the 681-nm chlorophyll absorption band; and variations over time in the EVI, GRND and NDVI. At the end of dry season, GV increased for Erisma uncinatum, while NPV increased for Chamaecrista xinguensis. For Manilkara huberi, the NPV first increased in the beginning of August and then decreased toward September with new foliage. Variations in red-edge position were not statistically significant between the species and across dates at the 95% confidence level. The camera data were affected by view-illumination effects, which reduced the SMA shade fraction over time. When MODIS data were corrected for these effects using the Multi-Angle Implementation of Atmospheric Correction Algorithm (MAIAC), we observed an EVI increase toward September that closely tracked the modeled LAI of mature leaves (3-5 months). Compared to the EVI, the GRND was a better indicator of leaf flushing because the modeled production of new leaves peaked in August and then declined in September following the GRND closely. While the EVI was more related to changes in mature leaf area, the GRND was more associated with new leaf flushing.

Cultivation of oyster mushroom Pleurotus ostreatus on date-palm leaves mixed with other agro-wastes in Saudi Arabia.

PubMed

Alananbeh, Kholoud M; Bouqellah, Nahla A; Al Kaff, Nadia S

2014-12-01

Promoting the use of agricultural waste is one of the newly prepared water and environment friendly agriculture strategies in the Kingdom of Saudi Arabia (KSA). The objective of this research was to study the efficiency of cultivating oyster mushroom (Pleurotus ostreatus) on date palm wastes mixed with other agricultural wastes available in KSA. Four agricultural wastes were mixed with date palm leaves at different ratios, with two supplements and three spawn rates were used. Wheat straw mixed with date palm at ratio of 25 (date palm): 75 (agro-waste) showed the best results in most of the parameters measured. Corn meal was superior over wheat bran as a supplement in all treatments. Parameter values increased with the increase of the spawn rate of P. ostreatus. Treatments with date palm leave wastes contained higher carbohydrates and fibers. No significant differences were found among the fruiting bodies produced on the different agro-wastes studied for the different proximates analyzed. Analyses of metal concentration showed that potassium was the highest in all the treatments tested followed by Na, Mg, Ca, and Zn. This is the first study that reported the success of growing oyster mushroom on date palm leaf wastes mixed with other agro-wastes obtainable in KSA.

Cultivation of oyster mushroom Pleurotus ostreatus on date-palm leaves mixed with other agro-wastes in Saudi Arabia

PubMed Central

Alananbeh, Kholoud M.; Bouqellah, Nahla A.; Al Kaff, Nadia S.

2014-01-01

Promoting the use of agricultural waste is one of the newly prepared water and environment friendly agriculture strategies in the Kingdom of Saudi Arabia (KSA). The objective of this research was to study the efficiency of cultivating oyster mushroom (Pleurotus ostreatus) on date palm wastes mixed with other agricultural wastes available in KSA. Four agricultural wastes were mixed with date palm leaves at different ratios, with two supplements and three spawn rates were used. Wheat straw mixed with date palm at ratio of 25 (date palm): 75 (agro-waste) showed the best results in most of the parameters measured. Corn meal was superior over wheat bran as a supplement in all treatments. Parameter values increased with the increase of the spawn rate of P. ostreatus. Treatments with date palm leave wastes contained higher carbohydrates and fibers. No significant differences were found among the fruiting bodies produced on the different agro-wastes studied for the different proximates analyzed. Analyses of metal concentration showed that potassium was the highest in all the treatments tested followed by Na, Mg, Ca, and Zn. This is the first study that reported the success of growing oyster mushroom on date palm leaf wastes mixed with other agro-wastes obtainable in KSA. PMID:25473372

The Membrane Transport System of the Guard Cell and Its Integration for Stomatal Dynamics1[CC-BY

PubMed Central

2017-01-01

Stomatal guard cells are widely recognized as the premier plant cell model for membrane transport, signaling, and homeostasis. This recognition is rooted in half a century of research into ion transport across the plasma and vacuolar membranes of guard cells that drive stomatal movements and the signaling mechanisms that regulate them. Stomatal guard cells surround pores in the epidermis of plant leaves, controlling the aperture of the pore to balance CO2 entry into the leaf for photosynthesis with water loss via transpiration. The position of guard cells in the epidermis is ideally suited for cellular and subcellular research, and their sensitivity to endogenous signals and environmental stimuli makes them a primary target for physiological studies. Stomata underpin the challenges of water availability and crop production that are expected to unfold over the next 20 to 30 years. A quantitative understanding of how ion transport is integrated and controlled is key to meeting these challenges and to engineering guard cells for improved water use efficiency and agricultural yields. PMID:28408539

The Membrane Transport System of the Guard Cell and Its Integration for Stomatal Dynamics.

PubMed

Jezek, Mareike; Blatt, Michael R

2017-06-01

Stomatal guard cells are widely recognized as the premier plant cell model for membrane transport, signaling, and homeostasis. This recognition is rooted in half a century of research into ion transport across the plasma and vacuolar membranes of guard cells that drive stomatal movements and the signaling mechanisms that regulate them. Stomatal guard cells surround pores in the epidermis of plant leaves, controlling the aperture of the pore to balance CO 2 entry into the leaf for photosynthesis with water loss via transpiration. The position of guard cells in the epidermis is ideally suited for cellular and subcellular research, and their sensitivity to endogenous signals and environmental stimuli makes them a primary target for physiological studies. Stomata underpin the challenges of water availability and crop production that are expected to unfold over the next 20 to 30 years. A quantitative understanding of how ion transport is integrated and controlled is key to meeting these challenges and to engineering guard cells for improved water use efficiency and agricultural yields. © 2017 The author(s). All Rights Reserved.

Time-dependent, x-ray spectral unfolds and brightness temperatures for intense Li{sup +} ion beam-driven hohlraums

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

Fehl, D.L.; Chandler, G.A.; Biggs, F.

X-ray-producing hohlraums are being studied as indirect drives for Inertial Confinement Fusion targets. In a 1994 target series on the PBFAII accelerator, cylindrical hohlraum targets were heated by an intense Li{sup +} ion beam and viewed by an array of 13 time-resolved, filtered x-ray detectors (XRDs). The UFO unfold code and its suite of auxiliary functions were used extensively in obtaining time- resolved x-ray spectra and radiation temperatures from this diagnostic. UFO was also used to obtain fitted response functions from calibration data, to simulate data from blackbody x-ray spectra of interest, to determine the suitability of various unfolding parametersmore » (e.g., energy domain, energy partition, smoothing conditions, and basis functions), to interpolate the XRD signal traces, and to unfold experimental data. The simulation capabilities of the code were useful in understanding an anomalous feature in the unfolded spectra at low photon energies ({le} 100 eV). Uncertainties in the differential and energy-integrated unfolded spectra were estimated from uncertainties in the data. The time-history of the radiation temperature agreed well with independent calculations of the wall temperature in the hohlraum.« less

Unfolding mechanism of thrombin-binding aptamer revealed by molecular dynamics simulation and Markov State Model

NASA Astrophysics Data System (ADS)

Zeng, Xiaojun; Zhang, Liyun; Xiao, Xiuchan; Jiang, Yuanyuan; Guo, Yanzhi; Yu, Xinyan; Pu, Xuemei; Li, Menglong

2016-04-01

Thrombin-binding aptamer (TBA) with the sequence 5‧GGTTGGTGTGGTTGG3‧ could fold into G-quadruplex, which correlates with functionally important genomic regionsis. However, unfolding mechanism involved in the structural stability of G-quadruplex has not been satisfactorily elucidated on experiments so far. Herein, we studied the unfolding pathway of TBA by a combination of molecular dynamics simulation (MD) and Markov State Model (MSM). Our results revealed that the unfolding of TBA is not a simple two-state process but proceeds along multiple pathways with multistate intermediates. One high flux confirms some observations from NMR experiment. Another high flux exhibits a different and simpler unfolding pathway with less intermediates. Two important intermediate states were identified. One is similar to the G-triplex reported in the folding of G-quadruplex, but lack of H-bonding between guanines in the upper plane. More importantly, another intermediate state acting as a connector to link the folding region and the unfolding one, was the first time identified, which exhibits higher population and stability than the G-triplex-like intermediate. These results will provide valuable information for extending our understanding the folding landscape of G-quadruplex formation.

Characterization of the Protein Unfolding Processes Induced by Urea and Temperature

PubMed Central

Rocco, Alessandro Guerini; Mollica, Luca; Ricchiuto, Piero; Baptista, António M.; Gianazza, Elisabetta; Eberini, Ivano

2008-01-01

Correct folding is critical for the biological activities of proteins. As a contribution to a better understanding of the protein (un)folding problem, we studied the effect of temperature and of urea on peptostreptococcal Protein L destructuration. We performed standard molecular dynamics simulations at 300 K, 350 K, 400 K, and 480 K, both in 10 M urea and in water. Protein L followed at least two alternative unfolding pathways. Urea caused the loss of secondary structure acting preferentially on the β-sheets, while leaving the α-helices almost intact; on the contrary, high temperature preserved the β-sheets and led to a complete loss of the α-helices. These data suggest that urea and high temperature act through different unfolding mechanisms, and protein secondary motives reveal a differential sensitivity to various denaturant treatments. As further validation of our results, replica-exchange molecular dynamics simulations of the temperature-induced unfolding process in the presence of urea were performed. This set of simulations allowed us to compute the thermodynamical parameters of the process and confirmed that, in the configurational space of Protein L unfolding, both of the above pathways are accessible, although to a different relative extent. PMID:18065481

The Role of Binding Site on the Mechanical Unfolding Mechanism of Ubiquitin

NASA Astrophysics Data System (ADS)

Cao, Penghui; Yoon, Gwonchan; Tao, Weiwei; Eom, Kilho; Park, Harold S.

2015-03-01

We apply novel atomistic simulations based on potential energy surface exploration to investigate the constant force-induced unfolding of ubiquitin. At the experimentally-studied force clamping level of 100 pN, we find a new unfolding mechanism starting with the detachment between β5 and β3 involving the binding site of ubiquitin, the Ile44 residue. This new unfolding pathway leads to the discovery of new intermediate configurations, which correspond to the end-to-end extensions previously seen experimentally. More importantly, it demonstrates the novel finding that the binding site of ubiquitin can be responsible not only for its biological functions, but also its unfolding dynamics. We also report in contrast to previous single molecule constant force experiments that when the clamping force becomes smaller than about 300 pN, the number of intermediate configurations increases dramatically, where almost all unfolding events at 100 pN involve an intermediate configuration. By directly calculating the life times of the intermediate configurations from the height of the barriers that were crossed on the potential energy surface, we demonstrate that these intermediate states were likely not observed experimentally due to their lifetimes typically being about two orders of magnitude smaller than the experimental temporal resolution.

Reconstructing the Origin of Helianthus deserticola: Survival and Selection on the Desert Floor

PubMed Central

Gross, Briana L.; Kane, Nolan C.; Lexer, Christian; Ludwig, Fulco; Rosenthal, David M.; Donovan, Lisa A.; Rieseberg, Loren H.

2008-01-01

The diploid hybrid species Helianthus deserticola inhabits the desert floor, an extreme environment relative to its parental species Helianthus annuus and Helianthus petiolaris. Adaptation to the desert floor may have occurred via selection acting on transgressive, or extreme, traits in early hybrids between the parental species. We explored this possibility through a field experiment in the hybrid species’ native habitat using H. deserticola, H. annuus, H. petiolaris, and two populations of early-generation (BC2) hybrids between the parental species, which served as proxies for the ancestral genotype of the ancient hybrid species. Character expression was evaluated for each genotypic class. Helianthus deserticola was negatively transgressive for stem diameter, leaf area, and flowering date, and the latter two traits are likely to be advantageous in a desert environment. The BC2 hybrids contained a range of variation that overlapped these transgressive trait means, and an analysis of phenotypic selection revealed that some of the selective pressures on leaf size and flowering date, but not stem diameter, would move the BC2 population toward the H. deserticola phenotype. Thus, H. deserticola may have originated from habitat-mediated directional selection acting on hybrids between H. annuus and H. petiolaris in a desert environment. PMID:15278840

Assessment of cardiac time intervals using high temporal resolution real-time spiral phase contrast with UNFOLDed-SENSE.

PubMed

Kowalik, Grzegorz T; Knight, Daniel S; Steeden, Jennifer A; Tann, Oliver; Odille, Freddy; Atkinson, David; Taylor, Andrew; Muthurangu, Vivek

2015-02-01

To develop a real-time phase contrast MR sequence with high enough temporal resolution to assess cardiac time intervals. The sequence utilized spiral trajectories with an acquisition strategy that allowed a combination of temporal encoding (Unaliasing by fourier-encoding the overlaps using the temporal dimension; UNFOLD) and parallel imaging (Sensitivity encoding; SENSE) to be used (UNFOLDed-SENSE). An in silico experiment was performed to determine the optimum UNFOLD filter. In vitro experiments were carried out to validate the accuracy of time intervals calculation and peak mean velocity quantification. In addition, 15 healthy volunteers were imaged with the new sequence, and cardiac time intervals were compared to reference standard Doppler echocardiography measures. For comparison, in silico, in vitro, and in vivo experiments were also carried out using sliding window reconstructions. The in vitro experiments demonstrated good agreement between real-time spiral UNFOLDed-SENSE phase contrast MR and the reference standard measurements of velocity and time intervals. The protocol was successfully performed in all volunteers. Subsequent measurement of time intervals produced values in keeping with literature values and good agreement with the gold standard echocardiography. Importantly, the proposed UNFOLDed-SENSE sequence outperformed the sliding window reconstructions. Cardiac time intervals can be successfully assessed with UNFOLDed-SENSE real-time spiral phase contrast. Real-time MR assessment of cardiac time intervals may be beneficial in assessment of patients with cardiac conditions such as diastolic dysfunction. © 2014 Wiley Periodicals, Inc.

Structural origins of pH and ionic strength effects on protein stability. Acid denaturation of sperm whale apomyoglobin.

PubMed

Yang, A S; Honig, B

1994-04-15

A recently developed approach to calculate the pH dependence of protein stability from three-dimensional structure information is applied to the analysis of acid denaturation of sperm whale apomyoglobin. The finite difference Poisson-Boltzmann method is used to calculate pKa values and these are used to obtain titration curves for the folded protein as well as for compact intermediates. The total electrostatic free energy change involved in apomyoglobin unfolding is then evaluated. Calculations are carried out of the unfolding free energy of the native (N) and the compact intermediate (I) of apomyoglobin relative to the unfolded state (U) over a range of pH at various ionic strengths. The contributions from key ionizable groups to the unfolding process are discussed. For the acid-induced partial unfolding of apomyoglobin near pH 5, the transition from N to I is found to be driven by three histidines that are exposed when the B, C, D and E helices unfold. Similarly, the unfolding of the compact intermediate I consisting of the A, G and H helices is driven primarily by a few carboxylic acids with low pKa values in the compact state. This picture is in contrast to the view which attributes acid denaturation to electrostatic repulsion resulting from the build up of positive charge. In fact, charge-charge interactions in myoglobin are found to be attractive at all pH values where the protein unfolds. pH-dependent changes in these interactions contribute to acid denaturation but other electrostatic effects, such as hydrogen bonding and solvation, are important as well. The effect of increasing ionic strength on unfolding is attributed to the decrease of attractive charge-charge interactions which destabilize the N state relative to I, but stabilize the I state relative to U by reducing the pKa shifts of a few critical carboxylic acids. The I state is found to be more stable than U at neutral pH thus accounting for its presence as an intermediate on the protein folding pathway. Our results have implications for the origins of compact intermediates or "molten globule" states.

Slow Proton Transfer Coupled to Unfolding Explains the Puzzling Results of Single-Molecule Experiments on BBL, a Paradigmatic Downhill Folding Protein

PubMed Central

Cerminara, Michele; Campos, Luis A.; Ramanathan, Ravishankar; Muñoz, Victor

2013-01-01

A battery of thermodynamic, kinetic, and structural approaches has indicated that the small α-helical protein BBL folds-unfolds via the one-state downhill scenario. Yet, single-molecule fluorescence spectroscopy offers a more conflicting view. Single-molecule experiments at pH 6 show a unique half-unfolded conformational ensemble at mid denaturation, whereas other experiments performed at higher pH show a bimodal distribution, as expected for two-state folding. Here we use thermodynamic and laser T-jump kinetic experiments combined with theoretical modeling to investigate the pH dependence of BBL stability, folding kinetics and mechanism within the pH 6–11 range. We find that BBL unfolding is tightly coupled to the protonation of one of its residues with an apparent pKa of ∼7. Therefore, in chemical denaturation experiments around neutral pH BBL unfolds gradually, and also converts in binary fashion to the protonated species. Moreover, under the single-molecule experimental conditions (denaturant midpoint and 279 K), we observe that proton transfer is much slower than the ∼15 microseconds folding-unfolding kinetics of BBL. The relaxation kinetics is distinctly biphasic, and the overall relaxation time (i.e. 0.2–0.5 ms) becomes controlled by the proton transfer step. We then show that a simple theoretical model of protein folding coupled to proton transfer explains quantitatively all these results as well as the two sets of single-molecule experiments, including their more puzzling features. Interestingly, this analysis suggests that BBL unfolds following a one-state downhill folding mechanism at all conditions. Accordingly, the source of the bimodal distributions observed during denaturation at pH 7–8 is the splitting of the unique conformational ensemble of BBL onto two slowly inter-converting protonation species. Both, the unprotonated and protonated species unfold gradually (one-state downhill), but they exhibit different degree of unfolding at any given condition because the native structure is less stable for the protonated form. PMID:24205082

Developing and Delivering National-Scale Gridded Phenology Data Products

NASA Astrophysics Data System (ADS)

Marsh, L.; Crimmins, M.; Crimmins, T. M.; Gerst, K.; Rosemartin, A.; Switzer, J.; Weltzin, J. F.

2016-12-01

The USA National Phenology Network (USA-NPN; www.usanpn.org) is now producing and freely delivering daily maps and short-term forecasts of accumulated growing degree days and spring onset dates (based on the Extended Spring Indices) at fine spatial scale for the conterminous United States. These data products have utility for a wide range of natural resource planning and management applications, including scheduling invasive species and pest detection and control activities, determining planting dates, anticipating allergy outbreaks and planning agricultural harvest dates. Accumulated growing degree day (AGDD) maps were selected because accumulated temperature is a strong driver of phenological transitions in plants and animals, including leaf-out, flowering, fruit ripening and migration. The Extended Spring Indices (SI-x) are based on predictive climate models for lilac and honeysuckle leaf and bloom; they have been widely used to summarize changes in the timing of spring onset. The SI-x is used as a national indicator of climate change impacts by the US Global Change Research Program and the Environmental Protection Agency. The USA-NPN is a national-scale program that supports scientific advancement and decision-making by collecting, storing, and sharing phenology data and information. To best serve various audiences, the AGDD and SI-x gridded maps are available in various formats through a range of access tools, including the USA-NPN online visualization tool as well as industry standards compliant web services. We plan to expand the suite of gridded map products offered by the USA-NPN to include predictive maps of phenological transitions for additional plant and animal species at fine spatial and temporal resolution in the near future. USA-NPN invites you to use freely available daily and short-term forecast maps of accumulated growing degree days and spring onset dates at fine spatial scale for the conterminous United States.

Application of principal component analysis in protein unfolding: an all-atom molecular dynamics simulation study.

PubMed

Das, Atanu; Mukhopadhyay, Chaitali

2007-10-28

We have performed molecular dynamics (MD) simulation of the thermal denaturation of one protein and one peptide-ubiquitin and melittin. To identify the correlation in dynamics among various secondary structural fragments and also the individual contribution of different residues towards thermal unfolding, principal component analysis method was applied in order to give a new insight to protein dynamics by analyzing the contribution of coefficients of principal components. The cross-correlation matrix obtained from MD simulation trajectory provided important information regarding the anisotropy of backbone dynamics that leads to unfolding. Unfolding of ubiquitin was found to be a three-state process, while that of melittin, though smaller and mostly helical, is more complicated.

Application of principal component analysis in protein unfolding: An all-atom molecular dynamics simulation study

NASA Astrophysics Data System (ADS)

Das, Atanu; Mukhopadhyay, Chaitali

2007-10-01

We have performed molecular dynamics (MD) simulation of the thermal denaturation of one protein and one peptide—ubiquitin and melittin. To identify the correlation in dynamics among various secondary structural fragments and also the individual contribution of different residues towards thermal unfolding, principal component analysis method was applied in order to give a new insight to protein dynamics by analyzing the contribution of coefficients of principal components. The cross-correlation matrix obtained from MD simulation trajectory provided important information regarding the anisotropy of backbone dynamics that leads to unfolding. Unfolding of ubiquitin was found to be a three-state process, while that of melittin, though smaller and mostly helical, is more complicated.

Odijk excluded volume interactions during the unfolding of DNA confined in a nanochannel.

PubMed

Reifenberger, Jeffrey G; Cao, Han; Dorfman, Kevin D

2018-02-13

We report experimental data on the unfolding of human and E. coli genomic DNA molecules shortly after injection into a 45 nm nanochannel. The unfolding dynamics are deterministic, consistent with previous experiments and modeling in larger channels, and do not depend on the biological origin of the DNA. The measured entropic unfolding force per friction per unit contour length agrees with that predicted by combining the Odijk excluded volume with numerical calculations of the Kirkwood diffusivity of confined DNA. The time scale emerging from our analysis has implications for genome mapping in nanochannels, especially as the technology moves towards longer DNA, by setting a lower bound for the delay time before making a measurement.

Galling by Rhopalomyia solidaginis alters Solidago altissima architecture and litter nutrient dynamics in an old-field ecosystem

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

Crutsinger, Greg; Habenicht, Melissa N; Classen, Aimee T

2008-01-01

Plant-insect interactions can alter ecosystem processes, especially if the insects modify plant architecture, quality, or the quantity of leaf litter inputs. In this study, we investigated the interactions between the gall midge Rhopalomyia solidaginis and tall goldenrod, Solidago altissima, to quantify the degree to which the midge alters plant architecture and how the galls affect rates of litter decomposition and nutrient release in an old-field ecosystem. R. solidaginis commonly leads to the formation of a distinct apical rosette gall on S. altissima and approximately 15% of the ramets in a S. altissima patch were galled (range: 3-34%). Aboveground biomass ofmore » galled ramets was 60% higher and the leaf area density was four times greater on galled leaf tissue relative to the portions of the plant that were not affected by the gall. Overall decomposition rate constants did not differ between galled and ungalled leaf litter. However, leaf-litter mass loss was lower in galled litter relative to ungalled litter, which was likely driven by modest differences in initial litter chemistry; this effect diminished after 12 weeks of decomposition in the field. The proportion of N remaining was always higher in galled litter than in ungalled litter at each collection date indicating differential release of nitrogen in galled leaf litter. Several studies have shown that plant-insect interactions on woody species can alter ecosystem processes by affecting the quality or quantity of litter inputs. Our results illustrate how plant-insect interactions in an herbaceous species can affect ecosystem processes by altering the quality and quantity of litter inputs. Given that S. altissima dominates fields and roadsides and that R. solidaginis galls are highly abundant throughout eastern North America, these interactions are likely to be important for both the structure and function of old-field ecosystems.« less

Interactive effects of UV radiation and reduced precipitation on the seasonal leaf phenolic content/composition and the antioxidant activity of naturally growing Arbutus unedo plants.

PubMed

Nenadis, Nikolaos; Llorens, Laura; Koufogianni, Agathi; Díaz, Laura; Font, Joan; Gonzalez, Josep Abel; Verdaguer, Dolors

2015-12-01

The effects of UV radiation and rainfall reduction on the seasonal leaf phenolic content/composition and antioxidant activity of the Mediterranean shrub Arbutus unedo were studied. Naturally growing plants of A. unedo were submitted to 97% UV-B reduction (UVA), 95% UV-A+UV-B reduction (UV0) or near-ambient UV levels (UVBA) under two precipitation regimes (natural rainfall or 10-30% rainfall reduction). Total phenol, flavonol and flavanol contents, levels of eight phenols and antioxidant activity [DPPH(●) radical scavenging and Cu (II) reducing capacity] were measured in sun-exposed leaves at the end of four consecutive seasons. Results showed a significant seasonal variation in the leaf content of phenols of A. unedo, with the lowest values found in spring and the highest in autumn and/or winter. Leaf ontogenetic development and/or a possible effect of low temperatures in autumn/winter may account for such findings. Regardless of the watering regime and the sampling date, plant exposure to UV-B radiation decreased the total flavanol content of leaves, while it increased the leaf content in quercitrin (the most abundant quercetin derivative identified). By contrast, UV-A radiation increased the leaf content of theogallin, a gallic acid derivative. Other phenolic compounds (two quercetin derivatives, one of them being avicularin, and one kaempferol derivative, juglanin), as well as the antioxidant activity of the leaves, showed different responses to UV radiation depending on the precipitation regime. Surprisingly, reduced rainfall significantly decreased the total amount of quantified quercetin derivatives as well as the DPPH scavenging activity in A. unedo leaves. To conclude, present findings indicate that leaves of A. unedo can be a good source of antioxidants throughout the year, but especially in autumn and winter. Copyright © 2015 Elsevier B.V. All rights reserved.

Identification and molecular characterization of a trans-acting small interfering RNA producing locus regulating leaf rust responsive gene expression in wheat (Triticum aestivum L.).

PubMed

Dutta, Summi; Kumar, Dhananjay; Jha, Shailendra; Prabhu, Kumble Vinod; Kumar, Manish; Mukhopadhyay, Kunal

2017-11-01

A novel leaf rust responsive ta-siRNA-producing locus was identified in wheat showing similarity to 28S rRNA and generated four differentially expressing ta-siRNAs by phasing which targeted stress responsive genes. Trans-acting-small interfering RNAs (Ta-siRNAs) are plant specific molecules generally involved in development and are also stress responsive. Ta-siRNAs identified in wheat till date are all responsive to abiotic stress only. Wheat cultivation is severely affected by rusts and leaf rust particularly affects grain filling. This study reports a novel ta-siRNA producing locus (TAS) in wheat which is a segment of 28S ribosomal RNA but shows differential expression during leaf rust infestation. Four small RNA libraries prepared from wheat Near Isogenic Lines were treated with leaf rust pathogen and compared with untreated controls. A TAS with the ability to generate four ta-siRNAs by phasing events was identified along with the microRNA TamiR16 as the phase initiator. The targets of the ta-siRNAs included α-gliadin, leucine rich repeat, trans-membrane proteins, glutathione-S-transferase, and fatty acid desaturase among others, which are either stress responsive genes or are essential for normal growth and development of plants. Expression of the TAS, its generated ta-siRNAs, and their target genes were profiled at five different time points after pathogen inoculation of susceptible and resistant wheat isolines and compared with mock-inoculated controls. Comparative analysis of expression unveiled differential and reciprocal relationship as well as discrete patterns between susceptible and resistant isolines. The expression profiles of the target genes of the identified ta-siRNAs advocate more towards effector triggered susceptibility favouring pathogenesis. The study helps in discerning the functions of wheat genes regulated by ta-siRNAs in response to leaf rust.

Cassava Plants with a Depleted Cyanogenic Glucoside Content in Leaves and Tubers. Distribution of Cyanogenic Glucosides, Their Site of Synthesis and Transport, and Blockage of the Biosynthesis by RNA Interference Technology1

PubMed Central

Jørgensen, Kirsten; Bak, Søren; Busk, Peter Kamp; Sørensen, Charlotte; Olsen, Carl Erik; Puonti-Kaerlas, Johanna; Møller, Birger Lindberg

2005-01-01

Transgenic cassava (Manihot esculenta Crantz, cv MCol22) plants with a 92% reduction in cyanogenic glucoside content in tubers and acyanogenic (<1% of wild type) leaves were obtained by RNA interference to block expression of CYP79D1 and CYP79D2, the two paralogous genes encoding the first committed enzymes in linamarin and lotaustralin synthesis. About 180 independent lines with acyanogenic (<1% of wild type) leaves were obtained. Only a few of these were depleted with respect to cyanogenic glucoside content in tubers. In agreement with this observation, girdling experiments demonstrated that cyanogenic glucosides are synthesized in the shoot apex and transported to the root, resulting in a negative concentration gradient basipetal in the plant with the concentration of cyanogenic glucosides being highest in the shoot apex and the petiole of the first unfolded leaf. Supply of nitrogen increased the cyanogenic glucoside concentration in the shoot apex. In situ polymerase chain reaction studies demonstrated that CYP79D1 and CYP79D2 were preferentially expressed in leaf mesophyll cells positioned adjacent to the epidermis. In young petioles, preferential expression was observed in the epidermis, in the two first cortex cell layers, and in the endodermis together with pericycle cells and specific parenchymatic cells around the laticifers. These data demonstrate that it is possible to drastically reduce the linamarin and lotaustralin content in cassava tubers by blockage of cyanogenic glucoside synthesis in leaves and petioles. The reduced flux to the roots of reduced nitrogen in the form of cyanogenic glucosides did not prevent tuber formation. PMID:16126856

SU-D-201-04: Evaluation of Elekta Agility MLC Performance Using Statistical Process Control

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

Meyers, SM; Balderson, MJ; Letourneau, D

2016-06-15

Purpose: to evaluate the performance and stability of the Elekta Agility MLC model using an automated quality control (QC) test in combination with statistical process control tools. Methods: Leaf positions were collected daily for 11 Elekta units over 5–19 months using the automated QC test, which analyzes 23 MV images to determine the location of MLC leaves relative to the radiation isocenter. The leaf positions are measured at 5 nominal positions, and images are acquired at collimator 0° and 180° to capture all MLC leaves in the field-of-view. Leaf positioning accuracy was assessed using individual and moving range control charts.more » Control limits were recomputed following MLC recalibration (occurred 1–2 times for 4 units). Specification levels of ±0.5, ±1 and ±1.5mm were tested. The mean and range of duration between out-of-control and out-of-specification events were determined. Results: Leaf position varied little over time, as confirmed by very tight individual control limits (mean ±0.19mm, range 0.09–0.44). Mean leaf position error was −0.03mm (range −0.89–0.83). Due to sporadic out-of-control events, the mean in-control duration was 3.3 days (range 1–23). Data stayed within ±1mm specification for 205 days on average (range 3–372) and within ±1.5mm for the entire date range. Measurements stayed within ±0.5mm for 1 day on average (range 0–17); however, our MLC leaves were not calibrated to this level of accuracy. Conclusion: The Elekta Agility MLC model was found to perform with high stability, as evidenced by the tight control limits. The in-specification durations support the current recommendation of monthly MLC QC tests with a ±1mm tolerance. Future work is on-going to determine if Agility performance can be optimized further using high-frequency QC test results to drive recalibration frequency. Factors that can affect leaf positioning accuracy, including beam spot motion, leaf gain calibration, drifting leaves, and image artifacts, are under investigation.« less

Single-molecule protein unfolding and translocation by an ATP-fueled proteolytic machine

PubMed Central

Aubin-Tam, Marie-Eve; Olivares, Adrian O.; Sauer, Robert T.; Baker, Tania A.; Lang, Matthew J.

2011-01-01

All cells employ ATP-powered proteases for protein-quality control and regulation. In the ClpXP protease, ClpX is a AAA+ machine that recognizes specific protein substrates, unfolds these molecules, and then translocates the denatured polypeptide through a central pore and into ClpP for degradation. Here, we use optical-trapping nanometry to probe the mechanics of enzymatic unfolding and translocation of single molecules of a multidomain substrate. Our experiments demonstrate the capacity of ClpXP and ClpX to perform mechanical work under load, reveal very fast and highly cooperative unfolding of individual substrate domains, suggest a translocation step size of 5–8 amino acids, and support a power-stroke model of denaturation in which successful enzyme-mediated unfolding of stable domains requires coincidence between mechanical pulling by the enzyme and a transient stochastic reduction in protein stability. We anticipate that single-molecule studies of the mechanical properties of other AAA+ proteolytic machines will reveal many shared features with ClpXP. PMID:21496645

A new seasonal-deciduous spring phenology submodel in the Community Land Model 4.5: impacts on carbon and water cycling under future climate scenarios.

PubMed

Chen, Min; Melaas, Eli K; Gray, Josh M; Friedl, Mark A; Richardson, Andrew D

2016-11-01

A spring phenology model that combines photoperiod with accumulated heating and chilling to predict spring leaf-out dates is optimized using PhenoCam observations and coupled into the Community Land Model (CLM) 4.5. In head-to-head comparison (using satellite data from 2003 to 2013 for validation) for model grid cells over the Northern Hemisphere deciduous broadleaf forests (5.5 million km 2 ), we found that the revised model substantially outperformed the standard CLM seasonal-deciduous spring phenology submodel at both coarse (0.9 × 1.25°) and fine (1 km) scales. The revised model also does a better job of representing recent (decadal) phenological trends observed globally by MODIS, as well as long-term trends (1950-2014) in the PEP725 European phenology dataset. Moreover, forward model runs suggested a stronger advancement (up to 11 days) of spring leaf-out by the end of the 21st century for the revised model. Trends toward earlier advancement are predicted for deciduous forests across the whole Northern Hemisphere boreal and temperate deciduous forest region for the revised model, whereas the standard model predicts earlier leaf-out in colder regions, but later leaf-out in warmer regions, and no trend globally. The earlier spring leaf-out predicted by the revised model resulted in enhanced gross primary production (up to 0.6 Pg C yr -1 ) and evapotranspiration (up to 24 mm yr -1 ) when results were integrated across the study region. These results suggest that the standard seasonal-deciduous submodel in CLM should be reconsidered, otherwise substantial errors in predictions of key land-atmosphere interactions and feedbacks may result. © 2016 John Wiley & Sons Ltd.

Antimalarial activity of Syzygium guineense during early and established Plasmodium infection in rodent models.

PubMed

Tadesse, Solomon Asmamaw; Wubneh, Zewdu Birhanu

2017-01-05

In Ethiopia, the leaves of Syzygium guineense have been found useful for the prevention and cure of malaria, and demonstrated antiplasmodial activity in vitro. Nevertheless, no scientific study has been conducted to confirm its antimalarial activity in vivo. Therefore, the objective of the study was to evaluate the antimalarial effect of Syzygium guineense leaf extract in mice. Inoculation of the study mice was carried out by using the malaria parasite, Plasmodium berghei. The plant extract was prepared at 200, 400 and 600 mg/kg. Chloroquine and distilled water was administered to the positive and negative control groups respectively. Parameters like parasitaemia, survival time and body weight were determined following standard tests (4-day suppressive, Rane's and repository tests). Syzygium guineense crude leaf extract displayed considerable (p < 0.05) parasite suppression at doses of 600 and 400 mg/kg in a 4-day suppressive test with chemosuppressive value of 59.39 and 49.09% respectively. S. guineense crude leaf extract also showed dose-dependent schizontocidal activity in both the repository and curative tests. The extract also prevented body weight loss and prolonged survival date of mice significantly (P < 0.05) at the highest dose employed in the study. Qualitative chemical assay for S. guineense methanolic leaf extract revealed that the plant is endowed with different plant secondary metabolites exemplified by terpenoids, alkaloids, triterpenes, flavonoids, anthraquinones, tannins, glycosides, saponins and phenols. Syzygium guineense leaf extract possess antimalarial activity in mice. The test substance was found to be safe with no observable signs of toxicity in the study mice. The results of the present work confirmed the in vitro antiplasmodial finding and traditional claims in vivo in mice. Therefore, Syzygium guineense could be regarded as a potential source to develop safe, effective and affordable antimalarial agent.

Estimating Parameters in the Generalized Graded Unfolding Model: Sensitivity to the Prior Distribution Assumption and the Number of Quadrature Points Used.

ERIC Educational Resources Information Center

Roberts, James S.; Donoghue, John R.; Laughlin, James E.

The generalized graded unfolding model (J. Roberts, J. Donoghue, and J. Laughlin, 1998, 1999) is an item response theory model designed to unfold polytomous responses. The model is based on a proximity relation that postulates higher levels of expected agreement with a given statement to the extent that a respondent is located close to the…

Mechanisms of m-cresol induced protein aggregation studied using a model protein cytochrome c†

PubMed Central

Singh, Surinder M.; Hutchings, Regina L.; Mallela, Krishna M.G.

2014-01-01

Multi-dose protein formulations require an effective antimicrobial preservative (AP) to inhibit microbial growth during long-term storage of unused formulations. m-cresol is one such AP, but has been shown to cause protein aggregation. However, the fundamental physical mechanisms underlying such AP-induced protein aggregation are not understood. In this study, we used a model protein cytochrome c to identify the protein unfolding that triggers protein aggregation. m-cresol induced cytochrome c aggregation at preservative concentrations that are commonly used to inhibit microbial growth. Addition of m-cresol decreased the temperature at which the protein aggregated and increased the aggregation rate. However, m-cresol did not perturb the tertiary or secondary structure of cytochrome c. Instead, it populated an “invisible” partially unfolded intermediate where a local protein region around the methionine residue at position 80 was unfolded. Stabilizing the Met80 region drastically decreased the protein aggregation, which conclusively shows that this local protein region acts as an aggregation “hot-spot”. Based on these results, we propose that APs induce protein aggregation by partial rather than global unfolding. Because of the availability of site-specific probes to monitor different levels of protein unfolding, cytochrome c provided a unique advantage in characterizing the partial protein unfolding that triggers protein aggregation. PMID:21229618

Temperature-induced unfolding of epidermal growth factor (EGF): insight from molecular dynamics simulation

PubMed Central

Yan, Chunli; Pattani, Varun; Tunnell, James W.; Ren, Pengyu

2010-01-01

Thermal disruption of protein structure and function is a potentially powerful therapeutic vehicle. With the emerging nanoparticle-targeting and femtosecond laser technology, it is possible to deliver heating locally to specific molecules. It is therefore important to understand how fast a protein can unfold or lose its function at high temperatures, such as near the water boiling point. In this study, the thermal damage of EGF was investigated by combining the replica exchange (136 replicas) and conventional molecular dynamics simulations. The REMD simulation was employed to rigorously explore the free energy landscape of EGF unfolding. Interestingly, besides the native and unfolded states, we also observed a distinct molten globule (MG) state that retained substantial amount of native contacts. Based on the understanding that which the unfolding of EGF is a three-state process, we have examined the unfolding kinetics of EGF (N→ MG→h multiple 20-ns conventional MD simulations. The Arrhenius prefactors and activation energy barriers determined from the simulation are within the range of previously studied proteins. In contrast to the thermal damage of cells and tissues which take place on the time scale of seconds to hours at relatively low temperatures, the denaturation of proteins occur in nanoseconds when the temperature of heat bath approaches the boiling point. PMID:20466569

Microsecond Unfolding Kinetics of Sheep Prion Protein Reveals an Intermediate that Correlates with Susceptibility to Classical Scrapie

PubMed Central

Chen, Kai-Chun; Xu, Ming; Wedemeyer, William J.; Roder, Heinrich

2011-01-01

The microsecond folding and unfolding kinetics of ovine prion proteins (ovPrP) were measured under various solution conditions. A fragment comprising residues 94–233 of the full-length ovPrP was studied for four variants with differing susceptibilities to classical scrapie in sheep. The observed biexponential unfolding kinetics of ovPrP provides evidence for an intermediate species. However, in contrast to previous results for human PrP, there is no evidence for an intermediate under refolding conditions. Global analysis of the kinetic data, based on a sequential three-state mechanism, quantitatively accounts for all folding and unfolding data as a function of denaturant concentration. The simulations predict that an intermediate accumulates under both folding and unfolding conditions, but is observable only in unfolding experiments because the intermediate is optically indistinguishable from the native state. The relative population of intermediates in two ovPrP variants, both transiently and under destabilizing equilibrium conditions, correlates with their propensities for classical scrapie. The variant susceptible to classical scrapie has a larger population of the intermediate state than the resistant variant. Thus, the susceptible variant should be favored to undergo the PrPC to PrPSc conversion and oligomerization. PMID:21889460

A hypothesis to reconcile the physical and chemical unfolding of proteins

PubMed Central

de Oliveira, Guilherme A. P.; Silva, Jerson L.

2015-01-01

High pressure (HP) or urea is commonly used to disturb folding species. Pressure favors the reversible unfolding of proteins by causing changes in the volumetric properties of the protein–solvent system. However, no mechanistic model has fully elucidated the effects of urea on structure unfolding, even though protein–urea interactions are considered to be crucial. Here, we provide NMR spectroscopy and 3D reconstructions from X-ray scattering to develop the “push-and-pull” hypothesis, which helps to explain the initial mechanism of chemical unfolding in light of the physical events triggered by HP. In studying MpNep2 from Moniliophthora perniciosa, we tracked two cooperative units using HP-NMR as MpNep2 moved uphill in the energy landscape; this process contrasts with the overall structural unfolding that occurs upon reaching a threshold concentration of urea. At subdenaturing concentrations of urea, we were able to trap a state in which urea is preferentially bound to the protein (as determined by NMR intensities and chemical shifts); this state is still folded and not additionally exposed to solvent [fluorescence and small-angle X-ray scattering (SAXS)]. This state has a higher susceptibility to pressure denaturation (lower p1/2 and larger ΔVu); thus, urea and HP share concomitant effects of urea binding and pulling and water-inducing pushing, respectively. These observations explain the differences between the molecular mechanisms that control the physical and chemical unfolding of proteins, thus opening up new possibilities for the study of protein folding and providing an interpretation of the nature of cooperativity in the folding and unfolding processes. PMID:25964355

Thermal unfolding of tetrameric melittin: comparison with the molten globule state of cytochrome c.

PubMed Central

Hagihara, Y.; Oobatake, M.; Goto, Y.

1994-01-01

Whereas melittin at micromolar concentrations is unfolded under conditions of low salt at neutral pH, it transforms to a tetrameric alpha-helical structure under several conditions, such as high peptide concentration, high anion concentration, or alkaline pH. The anion- and pH-dependent stabilization of the tetrameric structure is similar to that of the molten globule state of several acid-denatured proteins, suggesting that tetrameric melittin might be a state similar to the molten globule state. To test this possibility, we studied the thermal unfolding of tetrameric melittin using far-UV CD and differential scanning calorimetry. The latter technique revealed a broad but distinct heat absorption peak. The heat absorption curves were consistent with the unfolding transition observed by CD and were explainable by a 2-state transition mechanism between the tetrameric alpha-helical state and the monomeric unfolded state. From the peptide or salt-concentration dependence of unfolding, the heat capacity change upon unfolding was estimated to be 5 kJ (mol of tetramer)-1 K-1 at 30 degrees C and decreased with increasing temperature. The observed change in heat capacity was much smaller than that predicted from the crystallographic structure (9.2 kJ (mol of tetramer)-1 K-1), suggesting that the hydrophobic residues of tetrameric melittin in solution are exposed in comparison with the crystallographic structure. However, the results also indicate that the structure is more ordered than that of a typical molten globule state. We consider that the conformation is intermediate between the molten globule state and the native state of globular proteins. PMID:7833804

A neutron spectrum unfolding computer code based on artificial neural networks

NASA Astrophysics Data System (ADS)

Ortiz-Rodríguez, J. M.; Reyes Alfaro, A.; Reyes Haro, A.; Cervantes Viramontes, J. M.; Vega-Carrillo, H. R.

2014-02-01

The Bonner Spheres Spectrometer consists of a thermal neutron sensor placed at the center of a number of moderating polyethylene spheres of different diameters. From the measured readings, information can be derived about the spectrum of the neutron field where measurements were made. Disadvantages of the Bonner system are the weight associated with each sphere and the need to sequentially irradiate the spheres, requiring long exposure periods. Provided a well-established response matrix and adequate irradiation conditions, the most delicate part of neutron spectrometry, is the unfolding process. The derivation of the spectral information is not simple because the unknown is not given directly as a result of the measurements. The drawbacks associated with traditional unfolding procedures have motivated the need of complementary approaches. Novel methods based on Artificial Intelligence, mainly Artificial Neural Networks, have been widely investigated. In this work, a neutron spectrum unfolding code based on neural nets technology is presented. This code is called Neutron Spectrometry and Dosimetry with Artificial Neural networks unfolding code that was designed in a graphical interface. The core of the code is an embedded neural network architecture previously optimized using the robust design of artificial neural networks methodology. The main features of the code are: easy to use, friendly and intuitive to the user. This code was designed for a Bonner Sphere System based on a 6LiI(Eu) neutron detector and a response matrix expressed in 60 energy bins taken from an International Atomic Energy Agency compilation. The main feature of the code is that as entrance data, for unfolding the neutron spectrum, only seven rate counts measured with seven Bonner spheres are required; simultaneously the code calculates 15 dosimetric quantities as well as the total flux for radiation protection purposes. This code generates a full report with all information of the unfolding in the HTML format. NSDann unfolding code is freely available, upon request to the authors.

[Pregnancy date disk for inserting into prenatal records].

PubMed

Pluta, M; Dudenhausen, J W

1983-01-01

A pregnancy calculating disk - a loose-leaf insert for the antenatal record card--is intended to improve and relieve the prenatal service. A pregnancy calculating disk together with tables containing signs of risks and times for consultations helps towards speedy orientation in daily practice. The data displayed in graphic form gives the pregnant women information on the official guide-lines regarding recommended consultations and ultrasonic examinations during the course of a normal pregnancy.

Synchrony in the phenology of a culturally iconic spring flower

NASA Astrophysics Data System (ADS)

Sparks, Tim H.; Mizera, Tadeusz; Wójtowicz, Wanda; Tryjanowski, Piotr

2012-03-01

We examine the flowering phenology of the cultural iconic Spring Snowflake Leucojum vernum, a considerable tourist attraction, recorded from two sites in western Poland. Flowering dates at the two sites were closely correlated but about 6 days later at the more natural area. The end of flowering was associated with the start of canopy leafing. Early flowering was related to a longer flowering season which may benefit ecotourism under future climate warming.

Unfolding sphere size distributions with a density estimator based on Tikhonov regularization

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

Weese, J.; Korat, E.; Maier, D.

1997-12-01

This report proposes a method for unfolding sphere size distributions given a sample of radii that combines the advantages of a density estimator with those of Tikhonov regularization methods. The following topics are discusses in this report to achieve this method: the relation between the profile and the sphere size distribution; the method for unfolding sphere size distributions; the results based on simulations; and the experimental data comparison.

Warming and Chilling: Assessing Aspects of Changing Plant Ecology with Continental-scale Phenology

NASA Astrophysics Data System (ADS)

Schwartz, M. D.; Hanes, J. M.

2009-12-01

Many recent ecological studies have concentrated on the direct impacts of climate warming, such as modifications to seasonal plant and animal life cycle events (phenology). There are many examples, with most indicating earlier onset of spring plant growth and delayed onset of autumn senescence. However, the implication of continued warming for plant species’ chilling requirements has received comparatively less attention. Temperate zone woody plants often require a certain level of cool season "chilling" (accumulated time at temperatures below a specific threshold) to break dormancy and prepare to respond to springtime warming. Thus, the potential impacts of insufficient chilling must be included in a comprehensive assessment of plant species' responses to climate warming. Vegetation phenological data, when collected for specific plant species at continental-scale, can be used to extract information relating to the combined impacts of reduced chilling and warming on plant species physiology. In a recent study, we demonstrated that common lilac first leaf and first bloom phenology (collected from multiple locations in the western United States and matched with air temperature records) can estimate the species' chilling requirement (in this case 1748 chilling hours, below a base temperature of 7.2°C) and highlight the changing impact of warming on the plant's phenological response in light of that requirement. Specifically, when chilling is above the requirement, lilac first leaf dates advance at a rate of -5.0 days per 100 hour chilling accumulation reduction, and lilac first bloom dates advance at a rate of -4.2 days per 100 hour chilling accumulation reduction. In contrast, when chilling is below the requirement, the lilac event dates advance at a much reduced rate of -1.6 days per 100 hour reduction for first leaf date and -2.2 days per 100 hour reduction for first bloom date. Overall, these encouraging results for common lilac suggest that similar continental-scale phenological measurements could facilitate a better understanding of relationships among phenological response, springtime warming, and chilling requirements for other species. Further, it should be possible to address more detailed follow-up plant ecology questions in future studies using similar methodology. Example questions would include: 1) Are the chilling requirements for a species the same across its entire range? 2) Do species adapt to warming conditions by changing their chilling requirements? and 3) How much variation is there among species chilling requirements within the same community? Continental-scale phenological data sets are being developed by the USA National Phenology Network (http://www.usanpn.org), that will facilitate such investigations, and in turn be essential for understanding of (and eventually consideration of possible adaptations to) the coming impacts of climate warming on temperate plant communities. Additionally, these phenological data, because they provide plants species’ responses across large portions of species geographic ranges, will facilitate deeper understanding of the full range of plant-environment responses and consequently foster development of more robust phenological models.

Mechanistic insights on the responses of plant and ecosystem gas exchange to global environmental change: lessons from Biosphere 2.

PubMed

Gonzalez-Meler, Miquel A; Rucks, Jessica S; Aubanell, Gerard

2014-09-01

Scaling up leaf processes to canopy/ecosystem level fluxes is critical for examining feedbacks between vegetation and climate. Collectively, studies from Biosphere 2 Laboratory have provided important insight of leaf-to-ecosystem investigations of multiple environmental parameters that were not before possible in enclosed or field studies. B2L has been a testing lab for the applicability of new technologies such as spectral approaches to detect spatial and temporal changes in photosynthesis within canopies, or for the development of cavity ring-down isotope applications for ecosystem evapotranspiration. Short and long term changes in atmospheric CO2, drought or temperature allowed for intensive investigation of the interactions between photosynthesis and leaf, soil and ecosystem respiration. Experiments conducted in the rainforest biome have provided some of the most comprehensive dataset to date on the effects of climate change variables on tropical ecosystems. Results from these studies have been later corroborated in natural rainforest ecosystems and have improved the predictive capabilities of models that now show increased resilience of tropics to climate change. Studies of temperature and CO2 effects on ecosystem respiration and its leaf and soil components have helped reconsider the use of simple first-order kinetics for characterizing respiration in models. The B2L also provided opportunities to quantify the rhizosphere priming effect, or establish the relationships between net primary productivity, atmospheric CO2 and isoprene emissions. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Temporal Changes in Microscale Colonization of the Phylloplane by Aureobasidium pullulans

PubMed Central

McGrath, Molly J.; Andrews, John H.

2006-01-01

Colonization of apple leaves by the yeastlike fungus Aureobasidium pullulans was followed quantitatively and spatially at a microscale level throughout two growing seasons. Ten field leaves were sampled on 11 dates in 2003 and 15 dates in 2004. Using an A. pullulans-specific fluorescence in situ hybridization probe and epifluorescence microscopy, we enumerated total cells, swollen-cells and chlamydospores (SCC), and blastospores/mm2 on leaf features, including the midvein, other (smaller) veins, and the interveinal regions. By 7 July 2003 and 7 June 2004, the total numbers of A. pullulans cells/mm2 were significantly higher (P < 0.05) on the midvein and other veins than in the interveinal regions. This pattern remained consistent thereafter. The primary colonizing morphotype in all regions at all dates was the SCC form, although blastospores always occurred in low numbers. Occupancy was quantified based on the percentage of microscope fields of a particular leaf feature containing ≥1 A. pullulans cell. In general, as seasons progressed, the percent occupancy of features increased and, for most midvein and veinal features, approximated 100% at the end of both growing seasons. Except for early collections, when A. pullulans cell numbers were low, the percent occupancy of interveinal regions was lower than that of the midvein or other veinal regions. A. pullulans was distributed primarily as single cells throughout the seasons in interveinal regions. On the midvein and other veins, colonies of ≥4 cells developed over time, and more cells occurred in colonies than as singletons by August. Our results demonstrate that A. pullulans primarily colonizes veins, where populations appear to increase by growth in situ. This pattern is established early in the growing season and persists. PMID:16957250

Blueberry (Vaccinium corymbosum L.).

PubMed

Song, Guo-Qing

2015-01-01

Vaccinium consists of approximately 450 species, of which highbush blueberry (Vaccinium corymbosum) is one of the three major Vaccinium fruit crops (i.e., blueberry, cranberry, and lingonberry) domesticated in the twentieth century. In blueberry the adventitious shoot regeneration using leaf explants has been the most desirable regeneration system to date; Agrobacterium tumefaciens-mediated transformation is the major gene delivery method and effective selection has been reported using either the neomycin phosphotransferase II gene (nptII) or the bialaphos resistance (bar) gene as selectable markers. The A. tumefaciens-mediated transformation protocol described in this chapter is based on combining the optimal conditions for efficient plant regeneration, reliable gene delivery, and effective selection. The protocol has led to successful regeneration of transgenic plants from leaf explants of four commercially important highbush blueberry cultivars for multiple purposes, providing a powerful approach to supplement conventional breeding methods for blueberry by introducing genes of interest.

Spectral analysis of amazon canopy phenology during the dry season using a tower hyperspectral camera and modis observations

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

de Moura, Yhasmin Mendes; Galvão, Lênio Soares; Hilker, Thomas

The association between spectral reflectance and canopy processes remains challenging for quantifying large-scale canopy phenological cycles in tropical forests. In this paper, we used a tower-mounted hyperspectral camera in an eastern Amazon forest to assess how canopy spectral signals of three species are linked with phenological processes in the 2012 dry season. We explored different approaches to disentangle the spectral components of canopy phenology processes and analyze their variations over time using 17 images acquired by the camera. The methods included linear spectral mixture analysis (SMA); principal component analysis (PCA); continuum removal (CR); and first-order derivative analysis. In addition, threemore » vegetation indices potentially sensitive to leaf flushing, leaf loss and leaf area index (LAI) were calculated: the Enhanced Vegetation Index (EVI), Normalized Difference Vegetation Index (NDVI) and the entitled Green-Red Normalized Difference (GRND) index. We inspected also the consistency of the camera observations using Moderate Resolution Imaging Spectroradiometer (MODIS) and available phenological data on new leaf production and LAI of young, mature and old leaves simulated by a leaf demography-ontogeny model. The results showed a diversity of phenological responses during the 2012 dry season with related changes in canopy structure and greenness values. Because of the differences in timing and intensity of leaf flushing and leaf shedding, Erisma uncinatum, Manilkara huberi and Chamaecrista xinguensis presented different green vegetation (GV) and non-photosynthetic vegetation (NPV) SMA fractions; distinct PCA scores; changes in depth, width and area of the 681-nm chlorophyll absorption band; and variations over time in the EVI, GRND and NDVI. At the end of dry season, GV increased for Erisma uncinatum, while NPV increased for Chamaecrista xinguensis. For Manilkara huberi, the NPV first increased in the beginning of August and then decreased toward September with new foliage. Variations in red-edge position were not statistically significant between the species and across dates at the 95% confidence level. The camera data were affected by view-illumination effects, which reduced the SMA shade fraction over time. When MODIS data were corrected for these effects using the Multi-Angle Implementation of Atmospheric Correction Algorithm (MAIAC), we observed an EVI increase toward September that closely tracked the modeled LAI of mature leaves (3–5 months). Compared to the EVI, the GRND was a better indicator of leaf flushing because the modeled production of new leaves peaked in August and then declined in September following the GRND closely. Finally, while the EVI was more related to changes in mature leaf area, the GRND was more associated with new leaf flushing.« less

Spectral analysis of amazon canopy phenology during the dry season using a tower hyperspectral camera and modis observations

DOE PAGES

de Moura, Yhasmin Mendes; Galvão, Lênio Soares; Hilker, Thomas; ...

2017-09-01

The association between spectral reflectance and canopy processes remains challenging for quantifying large-scale canopy phenological cycles in tropical forests. In this paper, we used a tower-mounted hyperspectral camera in an eastern Amazon forest to assess how canopy spectral signals of three species are linked with phenological processes in the 2012 dry season. We explored different approaches to disentangle the spectral components of canopy phenology processes and analyze their variations over time using 17 images acquired by the camera. The methods included linear spectral mixture analysis (SMA); principal component analysis (PCA); continuum removal (CR); and first-order derivative analysis. In addition, threemore » vegetation indices potentially sensitive to leaf flushing, leaf loss and leaf area index (LAI) were calculated: the Enhanced Vegetation Index (EVI), Normalized Difference Vegetation Index (NDVI) and the entitled Green-Red Normalized Difference (GRND) index. We inspected also the consistency of the camera observations using Moderate Resolution Imaging Spectroradiometer (MODIS) and available phenological data on new leaf production and LAI of young, mature and old leaves simulated by a leaf demography-ontogeny model. The results showed a diversity of phenological responses during the 2012 dry season with related changes in canopy structure and greenness values. Because of the differences in timing and intensity of leaf flushing and leaf shedding, Erisma uncinatum, Manilkara huberi and Chamaecrista xinguensis presented different green vegetation (GV) and non-photosynthetic vegetation (NPV) SMA fractions; distinct PCA scores; changes in depth, width and area of the 681-nm chlorophyll absorption band; and variations over time in the EVI, GRND and NDVI. At the end of dry season, GV increased for Erisma uncinatum, while NPV increased for Chamaecrista xinguensis. For Manilkara huberi, the NPV first increased in the beginning of August and then decreased toward September with new foliage. Variations in red-edge position were not statistically significant between the species and across dates at the 95% confidence level. The camera data were affected by view-illumination effects, which reduced the SMA shade fraction over time. When MODIS data were corrected for these effects using the Multi-Angle Implementation of Atmospheric Correction Algorithm (MAIAC), we observed an EVI increase toward September that closely tracked the modeled LAI of mature leaves (3–5 months). Compared to the EVI, the GRND was a better indicator of leaf flushing because the modeled production of new leaves peaked in August and then declined in September following the GRND closely. Finally, while the EVI was more related to changes in mature leaf area, the GRND was more associated with new leaf flushing.« less

Measurement of protein unfolding/refolding kinetics and structural characterization of hidden intermediates by NMR relaxation dispersion

PubMed Central

Meinhold, Derrick W.; Wright, Peter E.

2011-01-01

Detailed understanding of protein function and malfunction hinges on the ability to characterize transiently populated states and the transitions between them. Here, we use 15N, , and 13CO NMR R2 relaxation dispersion to investigate spontaneous unfolding and refolding events of native apomyoglobin. Above pH 5.0, dispersion is dominated by processes involving fluctuations of the F-helix region, which is invisible in NMR spectra. Measurements of R2 dispersion for residues contacted by the F-helix region in the native (N) structure reveal a transient state formed by local unfolding of helix F and undocking from the protein core. A similar state was detected at pH 4.75–4.95 and determined to be an on-pathway intermediate (I1) in a linear three-state unfolding scheme (N⇆I1⇆MG) leading to a transiently populated molten globule (MG) state. The slowest steps in unfolding and refolding are N → I1 (36 s-1) and MG → I1 (26 s-1), respectively. Differences in chemical shift between N and I1 are very small, except in regions adjacent to helix F, showing that their core structures are similar. Chemical shift changes between the N and MG states, obtained from R2 dispersion, reveal that the transient MG state is structurally similar to the equilibrium MG observed previously at high temperature and low pH. Analysis of MG state chemical shifts shows the location of residual helical structure in the transient intermediate and identifies regions that unfold or rearrange into nonnative structure during the N → MG transition. The experiments also identify regions of energetic frustration that “crack” during unfolding and impede the refolding process. PMID:21562212

Spontaneous Unfolding-Refolding of Fibronectin Type III Domains Assayed by Thiol Exchange

PubMed Central

Shah, Riddhi; Ohashi, Tomoo; Erickson, Harold P.; Oas, Terrence G.

2017-01-01

Globular proteins are not permanently folded but spontaneously unfold and refold on time scales that can span orders of magnitude for different proteins. A longstanding debate in the protein-folding field is whether unfolding rates or folding rates correlate to the stability of a protein. In the present study, we have determined the unfolding and folding kinetics of 10 FNIII domains. FNIII domains are one of the most common protein folds and are present in 2% of animal proteins. FNIII domains are ideal for this study because they have an identical seven-strand β-sandwich structure, but they vary widely in sequence and thermodynamic stability. We assayed thermodynamic stability of each domain by equilibrium denaturation in urea. We then assayed the kinetics of domain opening and closing by a technique known as thiol exchange. For this we introduced a buried Cys at the identical location in each FNIII domain and measured the kinetics of labeling with DTNB over a range of urea concentrations. A global fit of the kinetics data gave the kinetics of spontaneous unfolding and refolding in zero urea. We found that the folding rates were relatively similar, ∼0.1–1 s−1, for the different domains. The unfolding rates varied widely and correlated with thermodynamic stability. Our study is the first to address this question using a set of domains that are structurally homologous but evolved with widely varying sequence identity and thermodynamic stability. These data add new evidence that thermodynamic stability correlates primarily with unfolding rate rather than folding rate. The study also has implications for the question of whether opening of FNIII domains contributes to the stretching of fibronectin matrix fibrils. PMID:27909052

Energetics, Ion and Water Binding of the Unfolding of AA/UU Base Pair Stacks and UAU/UAU Base Triplet Stacks in RNA.

PubMed

Carr, Carolyn E; Khutsishvili, Irine; Marky, Luis A

2018-06-22

Triplex formation occurs via interaction of a third strand with the major groove of double stranded nucleic acid, through Hoogsteen hydrogen bonding. In this work, we use a combination of temperature-dependent UV spectroscopy and differential scanning calorimetry to determine complete thermodynamic profiles for the unfolding of poly(rA)•poly(rU) (Duplex) and poly(rA)•2poly(rU) (Triplex). Our thermodynamic results are in good agreement with the much earlier work of Krakauer and Sturtevant using only UV melting techniques. The folding of these two helices yielded an uptake of ions, ΔnNa+ = 0.15 mol Na+/mol base-pair (Duplex) and 0.30 mol Na+/mole base-triplet (Triplex), which are consistent with their polymer behavior and the higher charge density parameter of triple helices. The osmotic stress technique yielded a release of structural water, ΔnW = 2 mol H2O/mol base-pair (Duplex unfolding into single strands) and an uptake of structural water, ΔnW = 2 mol H2O/mole base-pair (Triplex unfolding into Duplex and a single strand). However, an overall release of electrostricted waters is obtained for the unfolding of both complexes from pressure perturbation calorimetric experiments. In total, the ΔV values obtained for the unfolding of Triplex into Duplex and a single strand correspond to an immobilization of two structural waters and a release of three electrostricted waters. The ΔV values obtained for the unfolding of Duplex into two single strands correspond to the release of two structural waters and the immobilization of four electrostricted water molecules.

Unfolding of chondroitinase ABC Ι is dependent on thermodynamic driving force by kinetically rate constant-amplitude compensation: A stopped-flow fluorescence study.

PubMed

Shirdel, S Akram; Khalifeh, Khosrow; Ranjbar, Bijan; Golestani, Abolfazl; Khajeh, Khosro

2016-11-01

We had previously investigated the role of a loop on the activity and conformational stability of chondroitinase ABC Ι (cABC Ι) by constructing some representative mutants in which a network interaction around Asp 689 was manipulated. Here we extended our study by measuring the proteolytic resistance, long term and thermal stability as well as unfolding kinetics of these variants. Long term stability data at 4 and 25°C for 3 weeks indicates that all mutants remain considerably active at 4°C. Thermoinactivation rates for all variants shows that the wild type (WT) enzyme retained 50% of its activity after 2min keeping at 40°C, while L701T, H700N and H700N/L701T as conformationally stabilized variants, have slower inactivation rate. It was also found that compact and thermodynamically stabilized variants are more resistant to tryptolytic digestion. Also, kinetic curves of chemical unfolding of the enzyme variants from stopped-flow fluorescence measurements were best fitted into a three-exponential function with three rate constants and corresponding amplitudes. We found that the energy barrier of the fast unfolding phase is lower in stabilized variants; while the amplitude of this phase to the whole amplitude of the unfolding reaction is lower than that of destabilized variants, indicating more population of stabilized mutants unfold via slower unfolding phase. We concluded that the rate of local conformational change alone is not the same that is expected from global thermodynamic stability; however the corresponding amplitude can compensate the rate constant toward thermodynamic stability. Copyright © 2016 Elsevier Inc. All rights reserved.

A thiol probe for measuring unfolded protein load and proteostasis in cells.

PubMed

Chen, Moore Z; Moily, Nagaraj S; Bridgford, Jessica L; Wood, Rebecca J; Radwan, Mona; Smith, Trevor A; Song, Zhegang; Tang, Ben Zhong; Tilley, Leann; Xu, Xiaohong; Reid, Gavin E; Pouladi, Mahmoud A; Hong, Yuning; Hatters, Danny M

2017-09-07

When proteostasis becomes unbalanced, unfolded proteins can accumulate and aggregate. Here we report that the dye, tetraphenylethene maleimide (TPE-MI) can be used to measure cellular unfolded protein load. TPE-MI fluorescence is activated upon labelling free cysteine thiols, normally buried in the core of globular proteins that are exposed upon unfolding. Crucially TPE-MI does not become fluorescent when conjugated to soluble glutathione. We find that TPE-MI fluorescence is enhanced upon reaction with cellular proteomes under conditions promoting accumulation of unfolded proteins. TPE-MI reactivity can be used to track which proteins expose more cysteine residues under stress through proteomic analysis. We show that TPE-MI can report imbalances in proteostasis in induced pluripotent stem cell models of Huntington disease, as well as cells transfected with mutant Huntington exon 1 before the formation of visible aggregates. TPE-MI also detects protein damage following dihydroartemisinin treatment of the malaria parasites Plasmodium falciparum. TPE-MI therefore holds promise as a tool to probe proteostasis mechanisms in disease.Proteostasis is maintained through a number of molecular mechanisms, some of which function to protect the folded state of proteins. Here the authors demonstrate the use of TPE-MI in a fluorigenic dye assay for the quantitation of unfolded proteins that can be used to assess proteostasis on a cellular or proteome scale.

Comparative analysis of thermal unfolding simulations of RNA recognition motifs (RRMs) of TAR DNA-binding protein 43 (TDP-43).

PubMed

Prakash, Amresh; Kumar, Vijay; Meena, Naveen Kumar; Hassan, Md Imtaiyaz; Lynn, Andrew M

2018-01-10

TAR DNA-binding protein 43 (TDP-43) inclusions have been found in Amyotrophic lateral sclerosis (ALS) and several other neurodegenerative diseases. Many studies suggest the involvement of RNA recognition motifs (RRMs) in TDP-43 proteinopathy. To elucidate the structural stability and the unfolding dynamics of RRMs, we have carried out atomistic molecular dynamics simulations at two different temperatures (300 and 500 K). The simulations results indicate that there are distinct structural differences in the unfolding pathway between the two domains and RRM1 unfolds faster than RRM2 in accordance with the lower thermal stability found experimentally. The unfolding behaviors of secondary structures showed that the α-helix was more stable than β-sheet and structural rearrangements of β-sheets results in formation of additional α-helices. At higher temperature, RRM1 exhibit increased overall flexibility and unfolding than RRM2. The temperature-dependent free energy landscapes consist of multiple metastable states stabilized by non-native contacts and hydrogen bonds in RRM2, thus rendering the RRM2 more prone to misfolding. The structural rearrangements of RRM2 could lead to aberrant protein-protein interactions that may account for enhanced aggregation and toxicity of TDP-43. Our analysis, thus identify the structural and thermodynamic characteristics of the RRMs of TDP-43, which will serve to uncover molecular mechanisms and driving forces in TDP-43 misfolding and aggregation.

How cooperative are protein folding and unfolding transitions?

PubMed Central

Malhotra, Pooja

2016-01-01

Abstract A thermodynamically and kinetically simple picture of protein folding envisages only two states, native (N) and unfolded (U), separated by a single activation free energy barrier, and interconverting by cooperative two‐state transitions. The folding/unfolding transitions of many proteins occur, however, in multiple discrete steps associated with the formation of intermediates, which is indicative of reduced cooperativity. Furthermore, much advancement in experimental and computational approaches has demonstrated entirely non‐cooperative (gradual) transitions via a continuum of states and a multitude of small energetic barriers between the N and U states of some proteins. These findings have been instrumental towards providing a structural rationale for cooperative versus noncooperative transitions, based on the coupling between interaction networks in proteins. The cooperativity inherent in a folding/unfolding reaction appears to be context dependent, and can be tuned via experimental conditions which change the stabilities of N and U. The evolution of cooperativity in protein folding transitions is linked closely to the evolution of function as well as the aggregation propensity of the protein. A large activation energy barrier in a fully cooperative transition can provide the kinetic control required to prevent the accumulation of partially unfolded forms, which may promote aggregation. Nevertheless, increasing evidence for barrier‐less “downhill” folding, as well as for continuous “uphill” unfolding transitions, indicate that gradual non‐cooperative processes may be ubiquitous features on the free energy landscape of protein folding. PMID:27522064

Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war

DOE PAGES

Zheng, Xunhai; Pedersen, Lars C.; Gabel, Scott A.; ...

2016-01-14

Formation of the mature HIV-1 reverse transcriptase (RT) p66/p51 heterodimer requires subunit-specific processing of the p66/p66' homodimer precursor. Since the ribonuclease H (RH) domain contains an occult cleavage site located near its center, cleavage must occur either prior to folding or subsequent to unfolding. Recent NMR studies have identified a slow, subunit-specific RH domain unfolding process proposed to result from a residue tug-of-war between the polymerase and RH domains on the functionally inactive, p66' subunit. Here, we describe a structural comparison of the isolated RH domain with a domain swapped RH dimer that reveals several intrinsically destabilizing characteristics of themore » isolated domain that facilitate excursions of Tyr427 from its binding pocket and separation of helices B and D. These studies provide independent support for the subunit-selective RH domain unfolding pathway in which instability of the Tyr427 binding pocket facilitates its release followed by domain transfer, acting as a trigger for further RH domain destabilization and subsequent unfolding. As further support for this pathway, NMR studies demonstrate that addition of an RH active site-directed isoquinolone ligand retards the subunit-selective RH' domain unfolding behavior of the p66/p66' homodimer. As a result, this study demonstrates the feasibility of directly targeting RT maturation with therapeutics.« less

Unfolding the HIV-1 reverse transcriptase RNase H domain – how to lose a molecular tug-of-war

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

Zheng, Xunhai; Pedersen, Lars C.; Gabel, Scott A.

Formation of the mature HIV-1 reverse transcriptase (RT) p66/p51 heterodimer requires subunit-specific processing of the p66/p66' homodimer precursor. Since the ribonuclease H (RH) domain contains an occult cleavage site located near its center, cleavage must occur either prior to folding or subsequent to unfolding. Recent NMR studies have identified a slow, subunit-specific RH domain unfolding process proposed to result from a residue tug-of-war between the polymerase and RH domains on the functionally inactive, p66' subunit. Here, we describe a structural comparison of the isolated RH domain with a domain swapped RH dimer that reveals several intrinsically destabilizing characteristics of themore » isolated domain that facilitate excursions of Tyr427 from its binding pocket and separation of helices B and D. These studies provide independent support for the subunit-selective RH domain unfolding pathway in which instability of the Tyr427 binding pocket facilitates its release followed by domain transfer, acting as a trigger for further RH domain destabilization and subsequent unfolding. As further support for this pathway, NMR studies demonstrate that addition of an RH active site-directed isoquinolone ligand retards the subunit-selective RH' domain unfolding behavior of the p66/p66' homodimer. As a result, this study demonstrates the feasibility of directly targeting RT maturation with therapeutics.« less

Quantifying internal friction in unfolded and intrinsically disordered proteins with single-molecule spectroscopy

PubMed Central

Soranno, Andrea; Buchli, Brigitte; Nettels, Daniel; Cheng, Ryan R.; Müller-Späth, Sonja; Pfeil, Shawn H.; Hoffmann, Armin; Lipman, Everett A.; Makarov, Dmitrii E.; Schuler, Benjamin

2012-01-01

Internal friction, which reflects the “roughness” of the energy landscape, plays an important role for proteins by modulating the dynamics of their folding and other conformational changes. However, the experimental quantification of internal friction and its contribution to folding dynamics has remained challenging. Here we use the combination of single-molecule Förster resonance energy transfer, nanosecond fluorescence correlation spectroscopy, and microfluidic mixing to determine the reconfiguration times of unfolded proteins and investigate the mechanisms of internal friction contributing to their dynamics. Using concepts from polymer dynamics, we determine internal friction with three complementary, largely independent, and consistent approaches as an additive contribution to the reconfiguration time of the unfolded state. We find that the magnitude of internal friction correlates with the compactness of the unfolded protein: its contribution dominates the reconfiguration time of approximately 100 ns of the compact unfolded state of a small cold shock protein under native conditions, but decreases for more expanded chains, and approaches zero both at high denaturant concentrations and in intrinsically disordered proteins that are expanded due to intramolecular charge repulsion. Our results suggest that internal friction in the unfolded state will be particularly relevant for the kinetics of proteins that fold in the microsecond range or faster. The low internal friction in expanded intrinsically disordered proteins may have implications for the dynamics of their interactions with cellular binding partners. PMID:22492978

Quantifying internal friction in unfolded and intrinsically disordered proteins with single-molecule spectroscopy.

PubMed

Soranno, Andrea; Buchli, Brigitte; Nettels, Daniel; Cheng, Ryan R; Müller-Späth, Sonja; Pfeil, Shawn H; Hoffmann, Armin; Lipman, Everett A; Makarov, Dmitrii E; Schuler, Benjamin

2012-10-30

Internal friction, which reflects the "roughness" of the energy landscape, plays an important role for proteins by modulating the dynamics of their folding and other conformational changes. However, the experimental quantification of internal friction and its contribution to folding dynamics has remained challenging. Here we use the combination of single-molecule Förster resonance energy transfer, nanosecond fluorescence correlation spectroscopy, and microfluidic mixing to determine the reconfiguration times of unfolded proteins and investigate the mechanisms of internal friction contributing to their dynamics. Using concepts from polymer dynamics, we determine internal friction with three complementary, largely independent, and consistent approaches as an additive contribution to the reconfiguration time of the unfolded state. We find that the magnitude of internal friction correlates with the compactness of the unfolded protein: its contribution dominates the reconfiguration time of approximately 100 ns of the compact unfolded state of a small cold shock protein under native conditions, but decreases for more expanded chains, and approaches zero both at high denaturant concentrations and in intrinsically disordered proteins that are expanded due to intramolecular charge repulsion. Our results suggest that internal friction in the unfolded state will be particularly relevant for the kinetics of proteins that fold in the microsecond range or faster. The low internal friction in expanded intrinsically disordered proteins may have implications for the dynamics of their interactions with cellular binding partners.

Unfolding Kinetics of β-Lactoglobulin Induced by Surfactant and Denaturant: A Stopped-Flow/Fluorescence Study

PubMed Central

Viseu, Maria Isabel; Melo, Eduardo P.; Carvalho, Teresa Isabel; Correia, Raquel F.; Costa, Sílvia M. B.

2007-01-01

The β→α transition of β-lactoglobulin, a globular protein abundant in the milk of several mammals, is investigated in this work. This transition, induced by the cationic surfactant dodecyltrimethylammonium chloride (DTAC), is accompanied by partial unfolding of the protein. In this work, unfolding of bovine β-lactoglobulin in DTAC is compared with its unfolding induced by the chemical denaturant guanidine hydrochloride (GnHCl). The final protein states attained in the two media have quite different secondary structure: in DTAC the α-helical content increases, leading to the so-called α-state; in GnHCl the amount of ordered secondary-structure decreases, resulting in a random coil-rich final state (denatured, or D, state). To obtain information on both mechanistic routes, in DTAC and GnHCl, and to characterize intermediates, the kinetics of unfolding were investigated in the two media. Equilibrium and kinetic data show the partial accumulation of an on-pathway intermediate in each unfolding route: in DTAC, an intermediate (I1) with mostly native secondary structure but loose tertiary structure appears between the native (β) and α-states; in GnHCl, another intermediate (I2) appears between states β and D. Kinetic rate constants follow a linear Chevron-plot representation in GnHCl, but show a more complex mechanism in DTAC, which acts like a stronger binding species. PMID:17693475

3D hybrid tectono-stochastic modeling of naturally fractured reservoir: Application of finite element method and stochastic simulation technique

NASA Astrophysics Data System (ADS)

Gholizadeh Doonechaly, N.; Rahman, S. S.

2012-05-01

Simulation of naturally fractured reservoirs offers significant challenges due to the lack of a methodology that can utilize field data. To date several methods have been proposed by authors to characterize naturally fractured reservoirs. Among them is the unfolding/folding method which offers some degree of accuracy in estimating the probability of the existence of fractures in a reservoir. Also there are statistical approaches which integrate all levels of field data to simulate the fracture network. This approach, however, is dependent on the availability of data sources, such as seismic attributes, core descriptions, well logs, etc. which often make it difficult to obtain field wide. In this study a hybrid tectono-stochastic simulation is proposed to characterize a naturally fractured reservoir. A finite element based model is used to simulate the tectonic event of folding and unfolding of a geological structure. A nested neuro-stochastic technique is used to develop the inter-relationship between the data and at the same time it utilizes the sequential Gaussian approach to analyze field data along with fracture probability data. This approach has the ability to overcome commonly experienced discontinuity of the data in both horizontal and vertical directions. This hybrid technique is used to generate a discrete fracture network of a specific Australian gas reservoir, Palm Valley in the Northern Territory. Results of this study have significant benefit in accurately describing fluid flow simulation and well placement for maximal hydrocarbon recovery.

Effect of antimicrobial preservatives on partial protein unfolding and aggregation†

PubMed Central

Hutchings, Regina L.; Singh, Surinder M.; Cabello-Villegas, Javier; Mallela, Krishna M. G.

2014-01-01

One-third of protein formulations are multi-dose. These require antimicrobial preservatives (APs); however, some APs have been shown to cause protein aggregation. Our previous work on a model protein cytochrome c indicated that partial protein unfolding, rather than complete unfolding, triggers aggregation. Here, we examined the relative strength of five commonly used APs on such unfolding and aggregation, and explored whether stabilizing the aggregation “hot-spot” reduces such aggregation. All APs induced protein aggregation in the order m-cresol > phenol > benzyl alcohol > phenoxyethanol > chlorobutanol. All these enhanced the partial protein unfolding that includes a local region which was predicted to be the aggregation “hot-spot”. The extent of destabilization correlated with the extent of aggregation. Further, we show that stabilizing the “hot-spot” reduces aggregation induced by all five APs. These results indicate that m-cresol causes the most protein aggregation, whereas chlorobutanol causes the least protein aggregation. The same protein region acts as the “hot-spot” for aggregation induced by different APs, implying that developing strategies to prevent protein aggregation induced by one AP will also work for others. PMID:23169345

Amino acid substitutions affecting protein dynamics in eglin C do not affect heat capacity change upon unfolding.

PubMed

Gribenko, Alexey V; Keiffer, Timothy R; Makhatadze, George I

2006-08-01

The heat capacity change upon unfolding (deltaC(p)) is a thermodynamic parameter that defines the temperature dependence of the thermodynamic stability of proteins; however, physical basis of the heat capacity change is not completely understood. Although empirical surface area-based calculations can predict heat capacity changes reasonably well, accumulating evidence suggests that changes in hydration of those surfaces is not the only parameter contributing to the observed heat capacity changes upon unfolding. Because packing density in the protein interior is similar to that observed in organic crystals, we hypothesized that changes in protein dynamics resulting in increased rigidity of the protein structure might contribute to the observed heat capacity change upon unfolding. Using differential scanning calorimetry we characterized the thermodynamic behavior of a serine protease inhibitor eglin C and two eglin C variants with altered native state dynamics, as determined by NMR. We found no evidence of changes in deltaC(p) in either of the variants, suggesting that changes in rigidity do not contribute to the heat capacity change upon unfolding in this model system. Copyright 2006 Wiley-Liss, Inc.

Mechanical Unfolding Studies on Single-Domain SUMO and Multi-Domain Periplasmic Binding Proteins

NASA Astrophysics Data System (ADS)

Kotamarthi, Hema Chandra; Ainavarapu, Sri Rama Koti

Protein mechanics is a key component of many cellular and sub-cellular processes. The current review focuses on recent studies from our laboratory that probe the effect of sequence on the mechanical stability of structurally similar proteins and the unfolding mechanisms of multi-domain periplasmic binding proteins. Ubiquitin and small ubiquitin-related modifiers (SUMOs) are structurally similar and possess different mechanical stabilities, ubiquitin being stronger than SUMOs as revealed from their unfolding forces. These differences are plausibly due to the variation in number of inter-residue contacts. The unfolding potential widths determined from the pulling speed-dependent studies revealed that SUMOs are mechanically more flexible than ubiquitin. This flexibility of SUMOs plays a role in ligand binding and our single-molecule studies on SUMO interaction with SUMO binding motifs (SBMs) have shown that ligand binding decreases the SUMO flexibility and increases its mechanical stability. Studies on multi-domain periplasmic binding proteins have revealed that the unfolding energy landscape of these proteins is complex and they follow kinetic partitioning between two-state and multiple three-state pathways.

A Genome-Wide Survey of Date Palm Cultivars Supports Two Major Subpopulations in Phoenix dactylifera.

PubMed

Mathew, Lisa S; Seidel, Michael A; George, Binu; Mathew, Sweety; Spannagl, Manuel; Haberer, Georg; Torres, Maria F; Al-Dous, Eman K; Al-Azwani, Eman K; Diboun, Ilhem; Krueger, Robert R; Mayer, Klaus F X; Mohamoud, Yasmin Ali; Suhre, Karsten; Malek, Joel A

2015-05-08

The date palm (Phoenix dactylifera L.) is one of the oldest cultivated trees and is intimately tied to the history of human civilization. There are hundreds of commercial cultivars with distinct fruit shapes, colors, and sizes growing mainly in arid lands from the west of North Africa to India. The origin of date palm domestication is still uncertain, and few studies have attempted to document genetic diversity across multiple regions. We conducted genotyping-by-sequencing on 70 female cultivar samples from across the date palm-growing regions, including four Phoenix species as the outgroup. Here, for the first time, we generate genome-wide genotyping data for 13,000-65,000 SNPs in a diverse set of date palm fruit and leaf samples. Our analysis provides the first genome-wide evidence confirming recent findings that the date palm cultivars segregate into two main regions of shared genetic background from North Africa and the Arabian Gulf. We identify genomic regions with high densities of geographically segregating SNPs and also observe higher levels of allele fixation on the recently described X-chromosome than on the autosomes. Our results fit a model with two centers of earliest cultivation including date palms autochthonous to North Africa. These results adjust our understanding of human agriculture history and will provide the foundation for more directed functional studies and a better understanding of genetic diversity in date palm. Copyright © 2015 Mathew et al.

A Genome-Wide Survey of Date Palm Cultivars Supports Two Major Subpopulations in Phoenix dactylifera

PubMed Central

Mathew, Lisa S.; Seidel, Michael A.; George, Binu; Mathew, Sweety; Spannagl, Manuel; Haberer, Georg; Torres, Maria F.; Al-Dous, Eman K.; Al-Azwani, Eman K.; Diboun, Ilhem; Krueger, Robert R.; Mayer, Klaus F. X.; Mohamoud, Yasmin Ali; Suhre, Karsten; Malek, Joel A.

2015-01-01

The date palm (Phoenix dactylifera L.) is one of the oldest cultivated trees and is intimately tied to the history of human civilization. There are hundreds of commercial cultivars with distinct fruit shapes, colors, and sizes growing mainly in arid lands from the west of North Africa to India. The origin of date palm domestication is still uncertain, and few studies have attempted to document genetic diversity across multiple regions. We conducted genotyping-by-sequencing on 70 female cultivar samples from across the date palm–growing regions, including four Phoenix species as the outgroup. Here, for the first time, we generate genome-wide genotyping data for 13,000–65,000 SNPs in a diverse set of date palm fruit and leaf samples. Our analysis provides the first genome-wide evidence confirming recent findings that the date palm cultivars segregate into two main regions of shared genetic background from North Africa and the Arabian Gulf. We identify genomic regions with high densities of geographically segregating SNPs and also observe higher levels of allele fixation on the recently described X-chromosome than on the autosomes. Our results fit a model with two centers of earliest cultivation including date palms autochthonous to North Africa. These results adjust our understanding of human agriculture history and will provide the foundation for more directed functional studies and a better understanding of genetic diversity in date palm. PMID:25957276

Unfolding energetics and stability of banana lectin.

PubMed

Gupta, Garima; Sinha, Sharmistha; Surolia, Avadhesha

2008-08-01

The unfolding pathway of banana lectin from Musa paradisiaca was determined by isothermal denaturation induced by the chaotrope GdnCl. The unfolding was found to be a reversible process. The data obtained by isothermal denaturation provided information on conformational stability of banana lectin. The high values of DeltaG of unfolding at various temperatures indicated the strength of intersubunit interactions. It was found that banana lectin is a very stable and denatures at high chaotrope concentrations only. The basis of the stability may be attributed to strong hydrogen bonds of the order 2.5-3.1 A at the dimeric interface along with the presence of water bridges. This is perhaps very unique example in proteins where subunit association is not a consequence of the predominance of hydrophobic interactions. (c) 2008 Wiley-Liss, Inc.

Mapping and monitoring of crop intensity, calendar and irrigation using multi-temporal MODIS data

NASA Astrophysics Data System (ADS)

Xiao, X.; Boes, S.; Mulukutla, G.; Proussevitch, A.; Routhier, M.

2005-12-01

Agriculture is the most extensive land use and water use on the Earth. Because of the diverse range of natural environments and human needs, agriculture is also the most complicated land use and water use system, which poses an enormous challenge to the scientific community, the public and decision-makers. Updated and geo-referenced information on crop intensity (number of crops per year), calendar (planting date, harvesting date) and irrigation is critically needed to better understand the impacts of agriculture on biogeochemical cycles (e.g., carbon, nitrogen, trace gases), water and climate dynamics. Here we present an effort to develop a novel approach for mapping and monitoring crop intensity, calendar and irrigation, using multi-temporal Moderate Resolution Imaging Spectroradiometer (MODIS) image data. Our algorithm employed three vegetation indices that are sensitive to the seasonal dynamics of leaf area index, light absorption by leaf chlorophyll and land surface water content. Our objective is to generate geospatial databases of crop intensity, calendar and irrigation at 500-m spatial resolution and at 8-day temporal resolution. In this presentation, we report a preliminary geospatial dataset of paddy rice crop intensity, calendar and irrigation in Asia, which is developed from the 8-day composite images of MODIS in 2002. The resultant dataset could be used in many applications, including hydrological and climate modeling.

Comment on ``Experimental Free Energy Reconstruction From Single-Molecule Force Spectroscopy Using Jarzynski's Equality''

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

Friddle, R W

2008-01-14

Harris, Song and Kiang [1] (HSK) describe their results on reconstructing the free energy profiles for both the stretch of the titin polymer, and the unfolding of an individual I27 domain. The new finding reported in [1] is the measurement of the free energy barrier (or activation energy) to unfolding the I27 domain. Due to a misinterpretation of the mechanics involved, the free energy surface (and thus the energy barrier) to unfolding the I27 domain was not measured.

The reversible two-state unfolding of a monocot mannose-binding lectin from garlic bulbs reveals the dominant role of the dimeric interface in its stabilization.

PubMed

Bachhawat, K; Kapoor, M; Dam, T K; Surolia, A

2001-06-19

Allium sativum agglutinin (ASAI) is a heterodimeric mannose-specific bulb lectin possessing two polypeptide chains of molecular mass 11.5 and 12.5 kDa. The thermal unfolding of ASAI, characterized by differential scanning calorimetry and circular dichroism, shows it to be highly reversible and can be defined as a two-state process in which the folded dimer is converted directly to the unfolded monomers (A2 if 2U). Its conformational stability has been determined as a function of temperature, GdnCl concentration, and pH using a combination of thermal and isothermal GdnCl-induced unfolding monitored by DSC, far-UV CD, and fluorescence, respectively. Analyses of these data yielded the heat capacity change upon unfolding (DeltaC(p) and also the temperature dependence of the thermodynamic parameters, namely, DeltaG, DeltaH, and DeltaS. The fit of the stability curve to the modified Gibbs-Helmholtz equation provides an estimate of the thermodynamic parameters DeltaH(g), DeltaS(g), and DeltaC(p) as 174.1 kcal x mol(-1), 0.512 kcal x mol(-1) x K(-1), and 3.41 kcal x mol(-1) x K(-1), respectively, at T(g) = 339.4 K. Also, the free energy of unfolding, DeltaG(s), at its temperature of maximum stability (T(s) = 293 K) is 13.13 kcal x mol(-1). Unlike most oligomeric proteins studied so far, the lectin shows excellent agreement between the experimentally determined DeltaC(p) (3.2 +/- 0.28 kcal x mol(-1) x K(-1)) and those evaluated from a calculation of its accessible surface area. This in turn suggests that the protein attains a completely unfolded state irrespective of the method of denaturation. The absence of any folding intermediates suggests the quaternary interactions to be the major contributor to the conformational stability of the protein, which correlates well with its X-ray structure. The small DeltaC(p) for the unfolding of ASAI reflects a relatively small, buried hydrophobic core in the folded dimeric protein.

Trehalose delays the reversible but not the irreversible thermal denaturation of cutinase.

PubMed

Baptista, R P; Cabral, J M; Melo, E P

2000-12-20

The effect of trehalose (0.5 M) on the thermal stability of cutinase in the alkaline pH range was studied. The thermal unfolding induced by increasing temperature was analyzed in the absence and in the presence of trehalose according to a two-state model (which assumes that only the folded and unfolded states of cutinase were present). Trehalose delays the reversible unfolding. The midpoint temperature of the unfolding transition (Tm) increases by 4.0 degrees C and 2. 6 degrees C at pH 9.2 and 10.5, respectively, in the presence of trehalose. At pH 9.2 the thermal unfolding occurs at higher temperatures (Tm is 52.6 degrees C compared to 42.0 degrees C at pH 10.5) and a refolding yield of around 80% was obtained upon cooling. This pH value was chosen to study the irreversible inactivation (long-term stability) of cutinase. Temperatures in the transition range from folded to unfolded state were selected and the rate constants of irreversible inactivation determined. Inactivation followed first-order kinetics and trehalose reduced the observed rate constants of inactivation, pointing to a stabilizing effect on the irreversible inactivation step of thermal denaturation. However, if the contribution of reversible unfolding on the irreversible inactivation of cutinase was taken into account, i.e., considering the fraction of cutinase molecules in the reversible unfolded conformation, the intrinsic rate constants can be calculated. Based on the intrinsic rate constants it was concluded that trehalose does not delay the irreversible inactivation. This conclusion was further supported by comparing the activation energy of the irreversible inactivation in the absence and in the presence of trehalose. The apparent activation energy in the absence and in the presence of trehalose were 67 and 99 Kcal/mol, respectively. The activation energy calculated from intrinsic rate constants was higher in the absence (30 Kcal/mol) than in the presence of trehalose (16 Kcal/mol), showing that kinetics of the irreversible inactivation step increased in the presence of trehalose. In fact, trehalose stabilized only the reversible step of thermal denaturation of cutinase.

Collision induced unfolding of isolated proteins in the gas phase: past, present, and future.

PubMed

Dixit, Sugyan M; Polasky, Daniel A; Ruotolo, Brandon T

2018-02-01

Rapidly characterizing the three-dimensional structures of proteins and the multimeric machines they form remains one of the great challenges facing modern biological and medical sciences. Ion mobility-mass spectrometry based techniques are playing an expanding role in characterizing these functional complexes, especially in drug discovery and development workflows. Despite this expansion, ion mobility-mass spectrometry faces many challenges, especially in the context of detecting small differences in protein tertiary structure that bear functional consequences. Collision induced unfolding is an ion mobility-mass spectrometry method that enables the rapid differentiation of subtly-different protein isoforms based on their unfolding patterns and stabilities. In this review, we summarize the modern implementation of such gas-phase unfolding experiments and provide an overview of recent developments in both methods and applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

Understanding disordered and unfolded proteins using single-molecule FRET and polymer theory.

PubMed

Hofmann, Hagen

2016-11-17

Understanding protein folding and the functional properties of intrinsically disordered proteins (IDPs) requires detailed knowledge of the forces that act in polypeptide chains. These forces determine the dimensions and dynamics of unfolded and disordered proteins and have been suggested to impact processes such as the coupled binding and folding of IDPs, or the rate of protein folding reactions. Much of the progress in understanding the physical and chemical properties of unfolded and intrinsically disordered polypeptide chains has been made possible by the recent developments in single-molecule fluorescence techniques. However, the interpretation of the experimental results requires concepts from polymer physics in order to be understood. Here, I review some of the theories used to describe the dimensions of unfolded polypeptide chains under varying solvent conditions together with their more recent application to experimental data.

Molecular dynamics study of unfolding of lysozyme in water and its mixtures with dimethyl sulfoxide.

PubMed

Sedov, Igor A; Magsumov, Timur I

2017-09-01

All-atom explicit solvent molecular dynamics was used to study the process of unfolding of hen egg white lysozyme in water and mixtures of water with dimethyl sulfoxide at different compositions. We have determined the kinetic parameters of unfolding at a constant temperature 450K. For each run, the time of disruption of the tertiary structure of lysozyme t u was defined as the moment when a certain structural criterion computed from the trajectory reaches its critical value. A good agreement is observed between the results obtained using several different criteria. The secondary structure according to DSSP calculations is found to be partially unfolded to the moment of disruption of tertiary structure, but some of its elements keep for a long time after that. The values of t u averaged over ten 30ns-long trajectories for each solvent composition are shown to decrease very rapidly with addition of dimethyl sulfoxide, and rather small amounts of dimethyl sulfoxide are found to change the pathway of unfolding. In pure water, despite the loss of tertiary contacts and disruption of secondary structure elements, the protein preserves its compact globular state at least over 130ns of simulation, while even at 5mol percents of dimethyl sulfoxide it loses its compactness within 30ns. The proposed methodology is a generally applicable tool to quantify the rate of protein unfolding in simulation studies. Copyright © 2017 Elsevier Inc. All rights reserved.

Interactions of urea with native and unfolded proteins: a volumetric study.

PubMed

Son, Ikbae; Shek, Yuen Lai; Tikhomirova, Anna; Baltasar, Eduardo Hidalgo; Chalikian, Tigran V

2014-11-26

We describe a statistical thermodynamic approach to analyzing urea-dependent volumetric properties of proteins. We use this approach to analyze our urea-dependent data on the partial molar volume and adiabatic compressibility of lysozyme, apocytochrome c, ribonuclease A, and α-chymotrypsinogen A. The analysis produces the thermodynamic properties of elementary urea-protein association reactions while also yielding estimates of the effective solvent-accessible surface areas of the native and unfolded protein states. Lysozyme and apocytochrome c do not undergo urea-induced transitions. The former remains folded, while the latter is unfolded between 0 and 8 M urea. In contrast, ribonuclease A and α-chymotrypsinogen A exhibit urea-induced unfolding transitions. Thus, our data permit us to characterize urea-protein interactions in both the native and unfolded states. We interpreted the urea-dependent volumetric properties of the proteins in terms of the equilibrium constant, k, and changes in volume, ΔV0, and compressibility, ΔKT0, for a reaction in which urea binds to a protein with a concomitant release of two waters of hydration to the bulk. Comparison of the values of k, ΔV0, and ΔKT0 with the similar data obtained on small molecules mimicking protein groups reveals lack of cooperative effects involved in urea-protein interactions. In general, the volumetric approach, while providing a unique characterization of cosolvent-protein interactions, offers a practical way for evaluating the effective solvent accessible surface area of biologically significant fully or partially unfolded polypeptides.

Search for Functional Flexible Regions in the G-protein Family: New Reading of the FoldUnfold Program.

PubMed

Galzitskaya, Oxana; Deryusheva, Eugenia; Machulin, Andrey; Nemashkalova, Ekaterina; Glyakina, Anna

2018-06-21

High prediction accuracy of flexible loops in different protein families is a challenge because of the crucial functions associated with these regions. Results of the currently available programs for prediction of loops vary from protein to protein. For prediction of flexible regions in the G-domain for 23 representatives of G-proteins with the known 3D structure we have used eight programs. The results of predictions demonstrate that the FoldUnfold program predicts better loop positions than the PONDR, RОNN, DisEMBL, IUPred, GlobPlot 2, FoldIndex, and MobiDB programs. When classifying the predicted loops (rigid/flexible) according to the Debye-Waller fluctuation factors, our data reveal the existing weak correlation between the B-factors and the average number of closed residues according to the FoldUnfold program; the percentage of overlapping characteristics (residue fold/unfold status) of the protein residues from the two methods is about 60-70%. According to the FoldUnfold program, for G-proteins with the posttranslational modifications, the surrounding binding site residues by disordered-promoting glycine and alanine residues conduces to a more flexible position of the binding sites for fatty acid, while methionine, cysteine and isoleucine residues provide more rigid binding sites. Thus, our research demonstrates additional possibilities of the FoldUnfold program for prediction of flexible regions and characteristics of individual residues in a different protein family. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Essential Oil of Aristolochia trilobata: Synthesis, Routes of Exposure, Acute Toxicity, Binary Mixtures and Behavioral Effects on Leaf-Cutting Ants.

PubMed

de Oliveira, Bruna Maria S; Melo, Carlisson R; Alves, Péricles B; Santos, Abraão A; Santos, Ane Caroline C; Santana, Alisson da S; Araújo, Ana Paula A; Nascimento, Pedro E S; Blank, Arie F; Bacci, Leandro

2017-02-25

Plants of the genus Aristolochia have been frequently reported as important medicinal plants. Despite their high bioactive potential, to date, there are no reports of their effects on leaf-cutting ants. Therefore, the present study aimed to evaluate the insecticidal activity of the essential oil of Aristolochia trilobata and its major components on Atta sexdens and Acromyrmex balzani , two species of leaf-cutting ants. The bioassays were performed regarding routes of exposure, acute toxicity, binary mixtures of the major components and behavioral effects. Twenty-five components were identified in the essential oil of A. trilobata using a gas chromatographic system equipped with a mass spectrometer and a flame ionization detector. The components found in higher proportions were sulcatyl acetate, limonene, p -cymene and linalool. The essential oil of A. trilobata and its individual major components were efficient against A. balzani and A. sexdens workers when applied by fumigation. These components showed fast and efficient insecticidal activity on ants. The components acted synergistically and additively on A. balzani and A. sexdens , respectively, and caused a strong repellency/irritability in the ants. Thus, our results demonstrate the great potential of the essential oil of A. trilobata and its major components for the development of new insecticides.

Microhabitat associations of a semi-terrestrial fish, Kryptolebias marmoratus (Poey 1880) in a mosquito-ditched mangrove forest, west-central Florida

USGS Publications Warehouse

Richards, Travis M.; Krebs, Justin M.; McIvor, Carole C.

2011-01-01

Mangrove rivulus (Kryptolebias marmoratus) is one of the few species of fish that is semi-terrestrial and able to use exposed intertidal and potentially supratidal habitats for prolonged periods of time. Based on previous work demonstrating frequent use of subterranean crab burrows as well as damp leaf litter and logs, we examined the microhabitat associations of rivulus in a mosquito-ditched mangrove forest on the Gulf coast of Florida near the northern limit of its distribution. We captured 161 rivulus on 20 dates between late April and mid-December 2007 using trench traps. Fish ranged in size from 7 to 35 mm SL. Peak abundance in mid-summer coincided with recruitment of a new year-class. The three study sites occurred within 0.5 km of one another, and experienced similar water temperatures and salinities. Nevertheless, they differed in their degree of tidal inundation, standing stock of leaf litter, and density of entrances to fiddler crab burrows. We consistently observed the highest mean catches of rivulus away from permanent subtidal waters of mosquito ditches, at intermediate relative elevations, and where leaf litter was locally abundant. Density of entrances to crab burrows was apparently unrelated to rivulus distribution or abundance in these forests.

Hyperspectral and thermal imaging of oilseed rape (Brassica napus) response to fungal species of the genus Alternaria.

PubMed

Baranowski, Piotr; Jedryczka, Malgorzata; Mazurek, Wojciech; Babula-Skowronska, Danuta; Siedliska, Anna; Kaczmarek, Joanna

2015-01-01

In this paper, thermal (8-13 µm) and hyperspectral imaging in visible and near infrared (VNIR) and short wavelength infrared (SWIR) ranges were used to elaborate a method of early detection of biotic stresses caused by fungal species belonging to the genus Alternaria that were host (Alternaria alternata, Alternaria brassicae, and Alternaria brassicicola) and non-host (Alternaria dauci) pathogens to oilseed rape (Brassica napus L.). The measurements of disease severity for chosen dates after inoculation were compared to temperature distributions on infected leaves and to averaged reflectance characteristics. Statistical analysis revealed that leaf temperature distributions on particular days after inoculation and respective spectral characteristics, especially in the SWIR range (1000-2500 nm), significantly differed for the leaves inoculated with A. dauci from the other species of Alternaria as well as from leaves of non-treated plants. The significant differences in leaf temperature of the studied Alternaria species were observed in various stages of infection development. The classification experiments were performed on the hyperspectral data of the leaf surfaces to distinguish days after inoculation and Alternaria species. The second-derivative transformation of the spectral data together with back-propagation neural networks (BNNs) appeared to be the best combination for classification of days after inoculation (prediction accuracy 90.5%) and Alternaria species (prediction accuracy 80.5%).

HyspIRI Measurements of Agricultural Systems in California: 2013-2015

NASA Astrophysics Data System (ADS)

Townsend, P. A.; Kruger, E. L.; Singh, A.; Jablonski, A. D.; Kochaver, S.; Serbin, S.

2015-12-01

During 2013-2015, NASA collected high-altitude AVIRIS hyperspectral and MASTER thermal infrared imagery across large swaths of California in support of the HyspIRI planning and prototyping activities. During these campaigns, we made extensive measurements of photosynthetic capacity—Vcmax and Jmax—and their temperature sensitivities across a range of sites, crop types and environmental conditions. Our objectives were to characterize the physiological diversity of agricultural vegetation in California and develop generalizable algorithms to map these physiological parameters across several image acquisitions, regardless of crop type and canopy temperatures. We employed AVIRIS imagery to scale and estimate the vegetation parameters and MASTER surface temperature to provide context, since physiology responds exponentially to leaf temperature. We demonstrate a segmentation approach to disentangling leaf and background soil temperature, and then illustrate our retrievals of Vcmax and Jmax during overflight conditions across a large number of the 2013-2015 HyspIRI acquisitions. Our results show >80% repeatability (R2) across split sample jack-knifing, with RMSEs within 15% of the range of our data. The approach was robust across crop types (e.g., grape, almond, pistachio, avocado, pomegranate, oats, peppers, citrus, date palm, alfalfa, melons, beets) and leaf temperatures. A global imaging spectroscopy system such as HyspIRI will offer unprecedented ability to monitor agricultural crop performance under widely varying surface conditions.

Simulation of urea-induced protein unfolding: a lesson from bovine β-lactoglobulin.

PubMed

Eberini, Ivano; Emerson, Andrew; Sensi, Cristina; Ragona, Laura; Ricchiuto, Piero; Pedretti, Alessandro; Gianazza, Elisabetta; Tramontano, Anna

2011-09-01

To investigate the molecular mechanisms involved in the very initial stages of protein unfolding, we carried out one long (1 μs) simulation of bovine β-lactoglobulin (BLG) together with three (500 ns) supporting MD runs, in which the unfolding conditions were produced by adding the osmolyte urea to the simulated systems and/or by increasing the thermal energy raising the temperature from 300 to 350 K. BLG was chosen, since it is a well-characterized model protein, for which structural and folding properties have been widely investigated by X-ray and NMR. MD trajectories were analyzed not only in terms of standard progress variables, such as backbone H-bonds, gyration radius width, secondary structure elements, but also through the scrutiny of interactions and dynamical behavior of specific key residues previously pointed out and investigated by NMR and belonging to a well known hydrophobic cluster. MD trajectories simulated in different unfolding conditions suggest that urea destabilizes BLG structure weakening protein::protein hydrophobic interactions and the hydrogen bond network. The early unfolding events, better observed at higher temperature, affect both secondary and tertiary structure of the protein. Copyright © 2011 Elsevier Inc. All rights reserved.

Unfolding of titin immunoglobulin domains by steered molecular dynamics simulation.

PubMed

Lu, H; Isralewitz, B; Krammer, A; Vogel, V; Schulten, K

1998-08-01

Titin, a 1-microm-long protein found in striated muscle myofibrils, possesses unique elastic and extensibility properties in its I-band region, which is largely composed of a PEVK region (70% proline, glutamic acid, valine, and lysine residue) and seven-strand beta-sandwich immunoglobulin-like (Ig) domains. The behavior of titin as a multistage entropic spring has been shown in atomic force microscope and optical tweezer experiments to partially depend on the reversible unfolding of individual Ig domains. We performed steered molecular dynamics simulations to stretch single titin Ig domains in solution with pulling speeds of 0.5 and 1.0 A/ps. Resulting force-extension profiles exhibit a single dominant peak for each Ig domain unfolding, consistent with the experimentally observed sequential, as opposed to concerted, unfolding of Ig domains under external stretching forces. This force peak can be attributed to an initial burst of backbone hydrogen bonds, which takes place between antiparallel beta-strands A and B and between parallel beta-strands A' and G. Additional features of the simulations, including the position of the force peak and relative unfolding resistance of different Ig domains, can be related to experimental observations.

Peripheral Protein Unfolding Drives Membrane Bending.

PubMed

Siaw, Hew Ming Helen; Raghunath, Gokul; Dyer, R Brian

2018-06-20

Dynamic modulation of lipid membrane curvature can be achieved by a number of peripheral protein binding mechanisms such as hy-drophobic insertion of amphipathic helices and membrane scaffolding. Recently, an alternative mechanism was proposed in which crowding of peripherally bound proteins induces membrane curvature through steric pressure generated by lateral collisions. This effect was enhanced using intrinsically disordered proteins that possess high hydrodynamic radii, prompting us to explore whether membrane bending can be triggered by the folding-unfolding transition of surface-bound proteins. We utilized histidine-tagged human serum albumin bound to Ni-NTA-DGS containing liposomes as our model system to test this hypothesis. We found that reduction of the disulfide bonds in the protein resulted in unfolding of HSA, which subsequently led to membrane tubule formation. The frequency of tubule formation was found to be significantly higher when the proteins were unfolded while being localized to a phase-separated domain as opposed to randomly distributed in fluid phase liposomes, indicating that the steric pressure generated from protein unfolding is directly responsible for membrane deformation. Our results are critical for the design of peripheral membrane protein-immobilization strategies and open new avenues for exploring mechanisms of membrane bending driven by conformational changes of peripheral membrane proteins.

The Unfolding MD Simulations of Cyclophilin: Analyzed by Surface Contact Networks and Their Associated Metrics

PubMed Central

Roy, Sourav; Basu, Sankar; Dasgupta, Dipak; Bhattacharyya, Dhananjay; Banerjee, Rahul

2015-01-01

Currently, considerable interest exists with regard to the dissociation of close packed aminoacids within proteins, in the course of unfolding, which could result in either wet or dry moltenglobules. The progressive disjuncture of residues constituting the hydrophobic core ofcyclophilin from L. donovani (LdCyp) has been studied during the thermal unfolding of the molecule, by molecular dynamics simulations. LdCyp has been represented as a surface contactnetwork (SCN) based on the surface complementarity (Sm) of interacting residues within themolecular interior. The application of Sm to side chain packing within proteins make it a very sensitive indicator of subtle perturbations in packing, in the thermal unfolding of the protein. Network based metrics have been defined to track the sequential changes in the disintegration ofthe SCN spanning the hydrophobic core of LdCyp and these metrics prove to be highly sensitive compared to traditional metrics in indicating the increased conformational (and dynamical) flexibility in the network. These metrics have been applied to suggest criteria distinguishing DMG, WMG and transition state ensembles and to identify key residues involved in crucial conformational/topological events during the unfolding process. PMID:26545107

The E. coli thioredoxin folding mechanism: the key role of the C-terminal helix.

PubMed

Vazquez, Diego S; Sánchez, Ignacio E; Garrote, Ana; Sica, Mauricio P; Santos, Javier

2015-02-01

In this work, the unfolding mechanism of oxidized Escherichia coli thioredoxin (EcTRX) was investigated experimentally and computationally. We characterized seven point mutants distributed along the C-terminal α-helix (CTH) and the preceding loop. The mutations destabilized the protein against global unfolding while leaving the native structure unchanged. Global analysis of the unfolding kinetics of all variants revealed a linear unfolding route with a high-energy on-pathway intermediate state flanked by two transition state ensembles TSE1 and TSE2. The experiments show that CTH is mainly unfolded in TSE1 and the intermediate and becomes structured in TSE2. Structure-based molecular dynamics are in agreement with these experiments and provide protein-wide structural information on transient states. In our model, EcTRX folding starts with structure formation in the β-sheet, while the protein helices coalesce later. As a whole, our results indicate that the CTH is a critical module in the folding process, restraining a heterogeneous intermediate ensemble into a biologically active native state and providing the native protein with thermodynamic and kinetic stability. Copyright © 2014 Elsevier B.V. All rights reserved.

Pressure-induced subunit dissociation and unfolding of dimeric beta-lactoglobulin.

PubMed Central

Valente-Mesquita, V L; Botelho, M M; Ferreira, S T

1998-01-01

Effects of hydrostatic pressure on dimeric beta-lactoglobulin A (beta-Lg) were investigated. Application of pressures of up to 3.5 kbar induced a significant red shift ( approximately 11 nm) and a 60% increase in intrinsic fluorescence emission of beta-Lg. These changes were very similar to those induced by guanidine hydrochloride, which caused subunit dissociation and unfolding of beta-Lg. A large hysteresis in the recovery of fluorescence parameters was observed upon decompression of beta-Lg. Pressure-induced dissociation and unfolding were not fully reversible, because of the formation of a nonnative intersubunit disulfide bond that hampered correct refolding of the dimer. Comparison between pressure dissociation/unfolding at 3 degrees C and 23 degrees C revealed a marked destabilization of beta-Lg at low temperature. The stability of beta-Lg toward pressure was significantly enhanced by 1 M NaCl, but not by glycerol (up to 20% v/v). These observations suggest that salt stabilization was not related to a general cosolvent effect, but may reflect charge screening. Interestingly, pressure-induced dissociation/unfolding was completely independent of beta-Lg concentration, in apparent violation of the law of mass action. Possible causes for this anomalous behavior are discussed. PMID:9649408

Potential effect of atmospheric warming on grapevine phenology and post-harvest heat accumulation across a range of climates

NASA Astrophysics Data System (ADS)

Hall, Andrew; Mathews, Adam J.; Holzapfel, Bruno P.

2016-09-01

Carbohydrates are accumulated within the perennial structure of grapevines when their production exceeds the requirements of reproduction and growth. The period between harvest and leaf-fall (the post-harvest period) is a key period for carbohydrate accumulation in relatively warmer grape-growing regions. The level of carbohydrate reserves available for utilisation in the following season has an important effect on canopy growth and yield potential and is therefore an important consideration in vineyard management. In a warming climate, the post-harvest period is lengthening and becoming warmer, evidenced through studies in wine regions worldwide that have correlated recent air temperature increases with changing grapevine phenology. Budbreak, flowering, veraison, and harvest have all been observed to be occurring earlier than in previous decades. Additionally, the final stage of the grapevine phenological cycle, leaf-fall, occurs later. This study explored the potential for increased post-harvest carbohydrate accumulation by modelling heat accumulation following harvest dates for the recent climate (1975-2004) and two warmer climate projections with mean temperature anomalies of +1.26 and +2.61 °C. Summaries of post-harvest heat accumulation between harvest and leaf-fall were produced for each of Australia's Geographical Indications (wine regions) to provide comparisons from the base temperatures to projected warmer conditions across a range of climates. The results indicate that for warmer conditions, all regions observe earlier occurring budbreak and harvest as well as increasing post-harvest growing degree days accumulation before leaf-fall. The level of increase varies depending upon starting climatic condition, with cooler regions experiencing the greatest change.

Corn response to climate stress detected with satellite-based NDVI time series

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

Wang, Ruoyu; Cherkauer, Keith; Bowling, Laura

Corn growth conditions and yield are closely dependent on climate variability. Leaf growth, measured as the leaf area index, can be used to identify changes in crop growth in response to climate stress. This research was conducted to capture patterns of spatial and temporal corn leaf growth under climate stress for the St. Joseph River watershed, in northeastern Indiana. Leaf growth is represented by the Normalized Difference Vegetative Index (NDVI) retrieved from multiple years (2000–2010) of Landsat 5 TM images. By comparing NDVI values for individual image dates with the derived normal curve, the response of crop growth to environmentalmore » factors is quantified as NDVI residuals. Regression analysis revealed a significant relationship between yield and NDVI residual during the pre-silking period, indicating that NDVI residuals reflect crop stress in the early growing period that impacts yield. Both the mean NDVI residuals and the percentage of image pixels where corn was under stress (risky pixel rate) are significantly correlated with water stress. Dry weather is prone to hamper potential crop growth, with stress affecting most of the observed corn pixels in the area. Oversupply of rainfall at the end of the growing season was not found to have a measurable effect on crop growth, while above normal precipitation earlier in the growing season reduces the risk of yield loss at the watershed scale. Furthermore, the spatial extent of stress is much lower when precipitation is above normal than under dry conditions, masking the impact of small areas of yield loss at the watershed scale.« less

Impact of physiology, structure and BRDF in hyperspectral time series of a Citrus orchard

NASA Astrophysics Data System (ADS)

Stuckens, J.; Dzikiti, S.; Verstraeten, W. W.; Verreynne, J. S.; Swinnen, R.; Coppin, P.

2010-05-01

Monitoring of plant production systems using remote sensing requires an understanding of the mechanisms in which physiological and structural changes as well as the quality and direction of incident light alter the measured canopy reflectance. Due to the evergreen nature of Citrus, the benefits of year-round monitoring of spectral changes are counterweighted by more subtle changes and seasonal trends than in other perennials. This study presents the results of a 14 months field measurement campaign in a commercial Citrus sinensis ‘Midknight Valencia' orchard in Wellington, Western Cape Province, South-Africa. Hyperspectral data were collected of canopy and leaf reflectance (350 - 2500 nm) of 16 representative trees at monthly intervals and supplemented with local climatology, orchard management records, sap stream, water potential and leaf and soil nutrient analysis. The aim of this research is to translate spectral changes and trends at the leaf and at canopy levels into physiological processes such as plant nutrient and carbohydrate balances and stress responses. Specific research questions include the spectral detection of flowering (date of anthesis, flowering intensity), fruit drop, fruit number and coloration, vegetative flushes, leaf senescence and drop and pruning. Attention is paid to the detection and the impact of sunburn (photo-damage). In order to separate physiological and structural changes from changes caused by seasonal changes in solar elevation during measurement time (bidirectional reflectance) a normalization function is constructed using simulated and measured data. Additional research is done to up-scale measurements from tree level to orchard level, which includes the tree variability, the influence of soil and weeds and different amounts of shading.

Corn response to climate stress detected with satellite-based NDVI time series

DOE PAGES

Wang, Ruoyu; Cherkauer, Keith; Bowling, Laura

2016-03-23

Corn growth conditions and yield are closely dependent on climate variability. Leaf growth, measured as the leaf area index, can be used to identify changes in crop growth in response to climate stress. This research was conducted to capture patterns of spatial and temporal corn leaf growth under climate stress for the St. Joseph River watershed, in northeastern Indiana. Leaf growth is represented by the Normalized Difference Vegetative Index (NDVI) retrieved from multiple years (2000–2010) of Landsat 5 TM images. By comparing NDVI values for individual image dates with the derived normal curve, the response of crop growth to environmentalmore » factors is quantified as NDVI residuals. Regression analysis revealed a significant relationship between yield and NDVI residual during the pre-silking period, indicating that NDVI residuals reflect crop stress in the early growing period that impacts yield. Both the mean NDVI residuals and the percentage of image pixels where corn was under stress (risky pixel rate) are significantly correlated with water stress. Dry weather is prone to hamper potential crop growth, with stress affecting most of the observed corn pixels in the area. Oversupply of rainfall at the end of the growing season was not found to have a measurable effect on crop growth, while above normal precipitation earlier in the growing season reduces the risk of yield loss at the watershed scale. Furthermore, the spatial extent of stress is much lower when precipitation is above normal than under dry conditions, masking the impact of small areas of yield loss at the watershed scale.« less

Influences of Plant Species, Season and Location on Leaf Endophytic Bacterial Communities of Non-Cultivated Plants

PubMed Central

Ding, Tao; Melcher, Ulrich

2016-01-01

Bacteria are known to be associated endophytically with plants. Research on endophytic bacteria has identified their importance in food safety, agricultural production and phytoremediation. However, the diversity of endophytic bacterial communities and the forces that shape their compositions in non-cultivated plants are largely uncharacterized. In this study, we explored the diversity, community structure, and dynamics of endophytic bacteria in different plant species in the Tallgrass Prairie Preserve of northern Oklahoma, USA. High throughput sequencing of amplified segments of bacterial rDNA from 81 samples collected at four sampling times from five plant species at four locations identified 335 distinct OTUs at 97% sequence similarity, representing 16 phyla. Proteobacteria was the dominant phylum in the communities, followed by the phyla Bacteriodetes and Actinobacteria. Bacteria from four classes of Proteobacteria were detected with Alphaproteobacteria as the dominant class. Analysis of molecular variance revealed that host plant species and collecting date had significant influences on the compositions of the leaf endophytic bacterial communities. The proportion of Alphaproteobacteria was much higher in the communities from Asclepias viridis than from other plant species and differed from month to month. The most dominant bacterial groups identified in LDA Effect Size analysis showed host-specific patterns, indicating mutual selection between host plants and endophytic bacteria and that leaf endophytic bacterial compositions were dynamic, varying with the host plant’s growing season in three distinct patterns. In summary, next generation sequencing has revealed variations in the taxonomic compositions of leaf endophytic bacterial communities dependent primarily on the nature of the plant host species. PMID:26974817

Influences of Plant Species, Season and Location on Leaf Endophytic Bacterial Communities of Non-Cultivated Plants.

PubMed

Ding, Tao; Melcher, Ulrich

2016-01-01

Bacteria are known to be associated endophytically with plants. Research on endophytic bacteria has identified their importance in food safety, agricultural production and phytoremediation. However, the diversity of endophytic bacterial communities and the forces that shape their compositions in non-cultivated plants are largely uncharacterized. In this study, we explored the diversity, community structure, and dynamics of endophytic bacteria in different plant species in the Tallgrass Prairie Preserve of northern Oklahoma, USA. High throughput sequencing of amplified segments of bacterial rDNA from 81 samples collected at four sampling times from five plant species at four locations identified 335 distinct OTUs at 97% sequence similarity, representing 16 phyla. Proteobacteria was the dominant phylum in the communities, followed by the phyla Bacteriodetes and Actinobacteria. Bacteria from four classes of Proteobacteria were detected with Alphaproteobacteria as the dominant class. Analysis of molecular variance revealed that host plant species and collecting date had significant influences on the compositions of the leaf endophytic bacterial communities. The proportion of Alphaproteobacteria was much higher in the communities from Asclepias viridis than from other plant species and differed from month to month. The most dominant bacterial groups identified in LDA Effect Size analysis showed host-specific patterns, indicating mutual selection between host plants and endophytic bacteria and that leaf endophytic bacterial compositions were dynamic, varying with the host plant's growing season in three distinct patterns. In summary, next generation sequencing has revealed variations in the taxonomic compositions of leaf endophytic bacterial communities dependent primarily on the nature of the plant host species.

Holocene and Last Interglacial climate of the Faroe Islands from sedimentary leaf wax hydrogen isotopes

NASA Astrophysics Data System (ADS)

Curtin, L.; D'Andrea, W. J.; de Wet, G.; Balascio, N.; Bradley, R. S.

2017-12-01

The climate of the North Atlantic region is extremely sensitive to changes in ocean and atmospheric circulation, and understanding past natural variability in North Atlantic climate provides important context for modern climate change. Here, we present Holocene and Eemian hydrogen isotope (δD) records from leaf waxes preserved in lacustrine sediments from the North Atlantic Faroe Islands and interpret them as a proxy for temperature and hydroclimate variability. In addition to helping to constrain the timing and amplitude of climate evolution during each of these interglacial periods, the data can be used to directly compare Eemian and Holocene climate using the same proxy from the same terrestrial location. Of the leaf waxes measured, the δD values of long-chain and mid-chain n-alkanes showed two different signals, which we interpret to represent leaf water δD values and lake water δD values, respectively. The δD values for long-chain and mid-chain fatty acids were most similar to the mid-chain n-alkanes, and likely represent a mixture of terrestrial and aquatic sources. Leaf wax-inferred δD values of precipitation during the early Holocene (10,000 to 8,000 cal yr BP) are 13‰ enriched compared to the remainder of the Holocene (after 8,000 cal yr BP), which show relatively stable values over time. Inferred lake water δD values decreased slowly over the late Holocene, suggesting a gradual transition to a wetter climate after 4,000 cal yr BP. At 2,000 cal yr BP there was a significant change in the distribution of leaf waxes that suggests a transition from shrubland to grassland, but which pre-dates the pollen evidence for this transition. The last interglacial period has been suggested as an analog for future climate conditions. We found that long-chain alkane δD values from the Eemian are most similar to the earliest Holocene, which corroborate previous pollen studies suggesting a warmer climate at the Faroe Islands during this period.

Accumulation of Pb and Cu heavy metals in sea water, sediment, and leaf and root tissue of Enhalus sp. in the seagrass bed of Banten Bay

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

Fauziah, Faiza, E-mail: faiza.fauziah@gmail.com; Choesin, Devi N., E-mail: faiza.fauziah@gmail.com

2014-03-24

Banten Bay in Indonesia is a coastal area which has been highly affected by human activity. Previous studies have reported the presence of lead (Pb) and copper (Cu) heavy metals in the seawater of this area. This study was conducted to measure the accumulation of Pb and Cu in seawater, sediment, leaf tissue, and root tissue of the seagrass species Enhalus sp. Sampling was conducted at two observation stations in Banten Bay: Station 1 (St.1) was located closer to the coastline and to industrial plants as source of pollution, while Station 2 (St.2) was located farther away offshore. At eachmore » station, three sampling points were established by random sampling. Field sampling was conducted at two different dates, i.e., on 29 May 2012 and 30 June 2012. Samples were processed by wet ashing using concentrated HNO{sub 3} acid and measured using Atomic Absorption Spectrometry (AAS). Accumulation of Pb was only detected in sediment samples in St.1, while Cu was detected in all samples. Average concentrations of Cu in May were as follows: sediment St.1 = 0.731 ppm, sediment St.2 = 0.383 ppm, seawater St.1 = 0.163 ppm, seawater St.2 = 0.174 ppm, leaf St.1 = 0.102 ppm, leaf St.2 = 0.132 ppm, root St.1= 0.139 ppm, and root St.2 = 0.075 ppm. Average measurements of Cu in June were: sediment St.1 = 0.260 ppm, leaf St.1 = 0.335 ppm, leaf St.2 = 0.301 ppm, root St.1= 0.047 ppm, and root St.2 = 0.060 ppm. In June, Cu was undetected in St.2 sediment and seawater at both stations. In May, Cu concentration in seawater exceeded the maximum allowable threshold for water as determined by the Ministry of the Environment. Spatial and temporal variation in Pb and Cu accumulation were most probably affected by distance from source and physical conditions of the environment (e.g., water current and mixing)« less

Residue solvent accessibilities in the unfolded polypeptide chain.

PubMed Central

Zielenkiewicz, P; Saenger, W

1992-01-01

The difference of solvent accessibilities in the native and unfolded states of the protein is used as a measure of the hydrophobic contribution to the free energy of folding. We present a new approximation of amino acids solvent accessibilities in the unfolded state based on the 1-ns molecular dynamics simulation of Ala-X-Ala tripeptides at a temperature of 368 K. The standard accessibility values averaged from the molecular dynamics study are significantly lower from those previously obtained by considering only selected conformations of Ala-X-Ala tripeptides. PMID:1489908

Real-time investigation of protein unfolding at an air–water interface at the 1 s time scale

PubMed Central

Yano, Yohko F.; Arakawa, Etsuo; Voegeli, Wolfgang; Matsushita, Tadashi

2013-01-01

Protein unfolding at an air–water interface has been demonstrated such that the X-ray reflectivity can be measured with an acquisition time of 1 s using a recently developed simultaneous multiple-angle–wavelength-dispersive X-ray reflectometer. This has enabled the electron density profile of the adsorbed protein molecules to be obtained in real time. A globular protein, lysozyme, adsorbed at the air–water interface is found to unfold into a flat shape within 1 s. PMID:24121352

Phenology from Landsat when data is scarce: Using MODIS and Dynamic Time-Warping to combine multi-year Landsat imagery to derive annual phenology curves

NASA Astrophysics Data System (ADS)

Baumann, Matthias; Ozdogan, Mutlu; Richardson, Andrew D.; Radeloff, Volker C.

2017-02-01

Green-leaf phenology describes the development of vegetation throughout a growing season and greatly affects the interaction between climate and the biosphere. Remote sensing is a valuable tool to characterize phenology over large areas but doing at fine- to medium resolution (e.g., with Landsat data) is difficult because of low numbers of cloud-free images in a single year. One way to overcome data availability limitations is to merge multi-year imagery into one time series, but this requires accounting for phenological differences among years. Here we present a new approach that employed a time series of a MODIS vegetation index data to quantify interannual differences in phenology, and Dynamic Time Warping (DTW) to re-align multi-year Landsat images to a common phenology that eliminates year-to-year phenological differences. This allowed us to estimate annual phenology curves from Landsat between 2002 and 2012 from which we extracted key phenological dates in a Monte-Carlo simulation design, including green-up (GU), start-of-season (SoS), maturity (Mat), senescence (Sen), end-of-season (EoS) and dormancy (Dorm). We tested our approach in eight locations across the United States that represented forests of different types and without signs of recent forest disturbance. We compared Landsat-based phenological transition dates to those derived from MODIS and ground-based camera data from the PhenoCam-network. The Landsat and MODIS comparison showed strong agreement. Dates of green-up, start-of-season and maturity were highly correlated (r 0.86-0.95), as were senescence and end-of-season dates (r > 0.85) and dormancy (r > 0.75). Agreement between the Landsat and PhenoCam was generally lower, but correlation coefficients still exceeded 0.8 for all dates. In addition, because of the high data density in the new Landsat time series, the confidence intervals of the estimated keydates were substantially lower than in case of MODIS and PhenoCam. Our study thus suggests that by exploiting multi-year Landsat imagery and calibrating it with MODIS data it is possible to describe green-leaf phenology at much finer spatial resolution than previously possible, highlighting the potential for fine scale phenology maps using the rich Landsat data archive over large areas.

Effect of pH, temperature, and salt on the stability of Escherichia coli- and Chinese hamster ovary cell-derived IgG1 Fc.

PubMed

Li, Cynthia H; Narhi, Linda O; Wen, Jie; Dimitrova, Mariana; Wen, Zai-qing; Li, Jenny; Pollastrini, Joseph; Nguyen, Xichdao; Tsuruda, Trace; Jiang, Yijia

2012-12-18

The circulation half-life of a potential therapeutic can be increased by fusing the molecule of interest (an active peptide, the extracellular domain of a receptor, an enzyme, etc.) to the Fc fragment of a monoclonal antibody. For the fusion protein to be a successful therapeutic, it must be stable to process and long-term storage conditions, as well as to physiological conditions. The stability of the Fc used is critical for obtaining a successful therapeutic protein. The effects of pH, temperature, and salt on the stabilities of Escherichia coli- and Chinese hamster ovary cell (CHO)-derived IgG1 Fc high-order structure were probed using a variety of biophysical techniques. Fc molecules derived from both E. coli and CHO were compared. The IgG1 Fc molecules from both sources (glycosylated and aglycosylated) are folded at neutral pH and behave similarly upon heat- and low pH-induced unfolding. The unfolding of both IgG1 Fc molecules occurs via a multistep unfolding process, with the tertiary structure and C(H)2 domain unfolding first, followed by changes in the secondary structure and C(H)3 domain. The acid-induced unfolding of IgG1 Fc molecules is only partially reversible, with the formation of high-molecular weight species. The CHO-derived Fc protein (glycosylated) is more compact (smaller hydrodynamic radius) than the E. coli-derived protein (aglycosylated) at neutral pH. Unfolding is dependent on pH and salt concentration. The glycosylated C(H)2 domain melts at a temperature 4-5 °C higher than that of the aglycosylated domain, and the low-pH-induced unfolding of the glycosylated Fc molecule occurs at a pH ~0.5 pH unit lower than that of the aglycosylated protein. The difference observed between E. coli- and CHO-derived Fc molecules primarily involves the C(H)2 domain, where the glycosylation of the Fc resides.

Conformational stability of pGEX-expressed Schistosoma japonicum glutathione S-transferase: a detoxification enzyme and fusion-protein affinity tag.

PubMed Central

Kaplan, W.; Hüsler, P.; Klump, H.; Erhardt, J.; Sluis-Cremer, N.; Dirr, H.

1997-01-01

A glutathione S-transferase (Sj26GST) from Schistosoma japonicum, which functions in the parasite's Phase II detoxification pathway, is expressed by the Pharmacia pGEX-2T plasmid and is used widely as a fusion-protein affinity tag. It contains all 217 residues of Sj26GST and an additional 9-residue peptide linker with a thrombin cleavage site at its C-terminus. Size-exclusion HPLC (SEC-HPLC) and SDS-PAGE studies indicate that purification of the homodimeric protein under nonreducing conditions results in the reversible formation of significant amounts of 160-kDa and larger aggregates without a loss in catalytic activity. The basis for oxidative aggregation can be ascribed to the high degree of exposure of the four cysteine residues per subunit. The conformational stability of the dimeric protein was studied by urea- and temperature-induced unfolding techniques. Fluorescence-spectroscopy, SEC-HPLC, urea- and temperature-gradient gel electrophoresis, differential scanning microcalorimetry, and enzyme activity were employed to monitor structural and functional changes. The unfolding data indicate the absence of thermodynamically stable intermediates and that the unfolding/refolding transition is a two-state process involving folded native dimer and unfolded monomer. The stability of the protein was found to be dependent on its concentration, with a delta G degree (H2O) = 26.0 +/- 1.7 kcal/mol. The strong relationship observed between the m-value and the size of the protein indicates that the amount of protein surface area exposed to solvent upon unfolding is the major structural determinant for the dependence of the protein's free energy of unfolding on urea concentration. Thermograms obtained by differential scanning microcalorimetry also fitted a two-state unfolding transition model with values of delta Cp = 7,440 J/mol per K, delta H = 950.4 kJ/mol, and delta S = 1,484 J/mol. PMID:9041642

Conformational stability of pGEX-expressed Schistosoma japonicum glutathione S-transferase: a detoxification enzyme and fusion-protein affinity tag.

PubMed

Kaplan, W; Hüsler, P; Klump, H; Erhardt, J; Sluis-Cremer, N; Dirr, H

1997-02-01

A glutathione S-transferase (Sj26GST) from Schistosoma japonicum, which functions in the parasite's Phase II detoxification pathway, is expressed by the Pharmacia pGEX-2T plasmid and is used widely as a fusion-protein affinity tag. It contains all 217 residues of Sj26GST and an additional 9-residue peptide linker with a thrombin cleavage site at its C-terminus. Size-exclusion HPLC (SEC-HPLC) and SDS-PAGE studies indicate that purification of the homodimeric protein under nonreducing conditions results in the reversible formation of significant amounts of 160-kDa and larger aggregates without a loss in catalytic activity. The basis for oxidative aggregation can be ascribed to the high degree of exposure of the four cysteine residues per subunit. The conformational stability of the dimeric protein was studied by urea- and temperature-induced unfolding techniques. Fluorescence-spectroscopy, SEC-HPLC, urea- and temperature-gradient gel electrophoresis, differential scanning microcalorimetry, and enzyme activity were employed to monitor structural and functional changes. The unfolding data indicate the absence of thermodynamically stable intermediates and that the unfolding/refolding transition is a two-state process involving folded native dimer and unfolded monomer. The stability of the protein was found to be dependent on its concentration, with a delta G degree (H2O) = 26.0 +/- 1.7 kcal/mol. The strong relationship observed between the m-value and the size of the protein indicates that the amount of protein surface area exposed to solvent upon unfolding is the major structural determinant for the dependence of the protein's free energy of unfolding on urea concentration. Thermograms obtained by differential scanning microcalorimetry also fitted a two-state unfolding transition model with values of delta Cp = 7,440 J/mol per K, delta H = 950.4 kJ/mol, and delta S = 1,484 J/mol.

High- and low-temperature unfolding of human high-density apolipoprotein A-2.

PubMed Central

Gursky, O.; Atkinson, D.

1996-01-01

Human plasma apolipoprotein A-2 (apoA-2) is the second major protein of the high-density lipoproteins that mediate the transport and metabolism of cholesterol. Using CD spectroscopy and differential scanning calorimetry, we demonstrate that the structure of lipid-free apoA-2 in neutral low-salt solutions is most stable at approximately 25 degrees C and unfolds reversibly both upon heating and cooling from 25 degrees C. High-temperature unfolding of apoA-2, monitored by far-UV CD, extends from 25-85 degrees C with midpoint Th = 56 +/- 2 degrees C and vant Hoff's enthalpy delta H(Th) = 17 +/- 2 kcal/mol that is substantially lower than the expected enthalpy of melting of the alpha-helical structure. This suggests low-cooperativity apoA-2 unfolding. The apparent free energy of apoA-2 stabilization inferred from the CD analysis of the thermal unfolding, delta G(app)(25 degrees) = 0.82 +/- 0.15 kcal/mol, agrees with the value determined from chemical denaturation. Enhanced low-temperature stability of apoA-2 observed upon increase in Na2HPO4 concentration from 0.3 mM to 50 mM or addition of 10% glycerol may be linked to reduced water activity. The close proximity of the heat and cold unfolding transitions, that is consistent with low delta G(app)(25 degrees), indicates that lipid-free apoA-2 has a substantial hydrophobic core but is only marginally stable under near-physiological solvent conditions. This suggests that in vivo apoA-2 transfer is unlikely to proceed via the lipid-free state. Low delta H(Th) and low apparent delta Cp approximately 0.52 kcal/mol.K inferred from the far-UV CD analysis of apoA-2 unfolding, and absence of tertiary packing interactions involving Tyr groups suggested by near-UV CD, are consistent with a molten globular-like state of lipid-free apoA-2. PMID:8880911

Identification of Residual Structure in the Unfolded State of Ribonuclease H1 from the Moderately Thermophilic Chlorobium tepidum: Comparison with Thermophilic and Mesophilic Homologues†

PubMed Central

Ratcliff, Kathleen; Marqusee, Susan

2010-01-01

Ribonucleases H from organisms that grow at different temperatures demonstrate a variable change in heat capacity upon unfolding (ΔC°P) [Ratcliff, K., et al. (2009) Biochemistry 48, 5890–5898]. This ΔC°P has been shown to correlate with a tolerance to higher temperatures and residual structure in the unfolded state of the thermophilic proteins. In the RNase H from Thermus thermophilus, the low ΔC°P has been shown to arise from the same region as the folding core of the protein, and mutagenic studies have shown that loss of a hydrophobic residue in this region can disrupt this residual unfolded state structure and result in a return to a more mesophile-like ΔC°P [Robic, S., et al. (2002) Protein Sci. 11, 381–389; Robic, S., et al. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 11345–11349]. To understand further how residual structure in the unfolded state is encoded in the sequences of these thermophilic proteins, we subjected the RNase H from Chlorobium tepidum to similar studies. Analysis of new chimeric proteins reveals that like T. thermophilus RNase H, the folding core of C. tepidum RNaseH plays an important role in the unfolded state of this protein. Mutagenesis studies, based on both a computational investigation of the hydrophobic networks in the core region and comparisons with similar studies on T. thermophilus RNase H, identify new residues involved in this residual structure and suggest that the residual structure in the unfolded state of C. tepidum RNase H is more restricted than that of T. thermophilus. We conclude that while the folding core region determines the thermophilic-like behavior of this family of proteins, the residue-specific details vary. PMID:20491485

Stabilization of an α/β-hydrolase by introducing proline residues: salicylic binding protein 2 from tobacco

PubMed Central

Huang, Jun; Jones, Bryan J.; Kazlauskas, Romas J.

2015-01-01

α/β-Hydrolases are important enzymes for biocatalysis, but their stability often limits their application. As a model α/β-hydrolase, we investigated a plant esterase, salicylic acid binding protein 2 (SABP2). SABP2 shows typical stability to urea (unfolding free energy 6.9±1.5 kcal/mol) and to heat inactivation (T1/215 min 49.2±0.5 °C). Denaturation in urea occurs in two steps, but heat inactivation occurs in a single step. The first unfolding step in urea eliminates catalytic activity. Surprisingly, we found that the first unfolding likely corresponds to the unfolding of the larger catalytic domain. Replacing selected amino acid residues with proline stabilized SABP2. Proline restricts the flexibility of the unfolded protein, thereby shifting the equilibrium toward the folded conformation. Seven locations for proline substitution were chosen either by amino acid sequence alignment with a more stable homolog or by targeting flexible regions in SABP2. Introducing proline in the catalytic domain stabilized SABP2 to the first unfolding in urea for three of five cases: L46P (+0.2 M urea), S70P (+0.1) and E215P (+0.9). Introducing proline in the cap domain did not (two of two cases), supporting the assignment that the first unfolding corresponds to the catalytic domain. Proline substitutions in both domains stabilized SABP2 to heat inactivation: L46P (ΔT1/215 min = +6.4 °C), S70P (+5.4), S115P (+1.8), S141P (+4.9), and E215P (+4.2). Combining substitutions did not further increase the stability to urea denaturation, but dramatically increased resistance to heat inactivation: L46P-S70P ΔT1/215 min = +25.7 °C. This straightforward proline substitution approach may also stabilize other α/β-hydrolases. PMID:26110207

Single-molecule studies of the Im7 folding landscape.

PubMed

Pugh, Sara D; Gell, Christopher; Smith, D Alastair; Radford, Sheena E; Brockwell, David J

2010-04-23

Under appropriate conditions, the four-helical Im7 (immunity protein 7) folds from an ensemble of unfolded conformers to a highly compact native state via an on-pathway intermediate. Here, we investigate the unfolded, intermediate, and native states populated during folding using diffusion single-pair fluorescence resonance energy transfer by measuring the efficiency of energy transfer (or proximity or P ratio) between pairs of fluorophores introduced into the side chains of cysteine residues placed in the center of helices 1 and 4, 1 and 3, or 2 and 4. We show that while the native states of each variant give rise to a single narrow distribution with high P values, the distributions of the intermediates trapped at equilibrium (denoted I(eqm)) are fitted by two Gaussian distributions. Modulation of the folding conditions from those that stabilize the intermediate to those that destabilize the intermediate enabled the distribution of lower P value to be assigned to the population of the unfolded ensemble in equilibrium with the intermediate state. The reduced stability of the I(eqm) variants allowed analysis of the effect of denaturant concentration on the compaction and breadth of the unfolded state ensemble to be quantified from 0 to 6 M urea. Significant compaction is observed as the concentration of urea is decreased in both the presence and absence of sodium sulfate, as previously reported for a variety of proteins. In the presence of Na(2)SO(4) in 0 M urea, the P value of the unfolded state ensemble approaches that of the native state. Concurrent with compaction, the ensemble displays increased peak width of P values, possibly reflecting a reduction in the rate of conformational exchange among iso-energetic unfolded, but compact conformations. The results provide new insights into the initial stages of folding of Im7 and suggest that the unfolded state is highly conformationally constrained at the outset of folding. (c) 2010 Elsevier Ltd. All rights reserved.

Thermal and urea-induced unfolding in T7 RNA polymerase: Calorimetry, circular dichroism and fluorescence study

PubMed Central

Griko, Yuri; Sreerama, Narasimha; Osumi-Davis, Patricia; Woody, Robert W.; Woody, A-Young Moon

2001-01-01

Structural changes in T7 RNA polymerase (T7RNAP) induced by temperature and urea have been studied over a wide range of conditions to obtain information about the structural organization and the stability of the enzyme. T7RNAP is a large monomeric enzyme (99 kD). Calorimetric studies of the thermal transitions in T7RNAP show that the enzyme consists of three cooperative units that may be regarded as structural domains. Interactions between these structural domains and their stability strongly depend on solvent conditions. The unfolding of T7RNAP under different solvent conditions induces a highly stable intermediate state that lacks specific tertiary interactions, contains a significant amount of residual secondary structure, and undergoes further cooperative unfolding at high urea concentrations. Circular dichroism (CD) studies show that thermal unfolding leads to an intermediate state that has increased β-sheet and reduced α-helix content relative to the native state. Urea-induced unfolding at 25°C reveals a two-step process. The first transition centered near 3 M urea leads to a plateau from 3.5 to 5.0 M urea, followed by a second transition centered near 6.5 M urea. The CD spectrum of the enzyme in the plateau region, which is similar to that of the enzyme thermally unfolded in the absence of urea, shows little temperature dependence from 15° to 60°C. The second transition leads to a mixture of poly(Pro)II and unordered conformations. As the temperature increases, the ellipticity at 222 nm becomes more negative because of conversion of poly(Pro)II to the unordered conformation. Near-ultraviolet CD spectra at 25°C at varying concentrations of urea are consistent with this picture. Both thermal and urea denaturation are irreversible, presumably because of processes that follow unfolding. PMID:11274475

Single-Molecule Studies of the Im7 Folding Landscape

PubMed Central

Pugh, Sara D.; Gell, Christopher; Smith, D. Alastair; Radford, Sheena E.; Brockwell, David J.

2010-01-01

Under appropriate conditions, the four-helical Im7 (immunity protein 7) folds from an ensemble of unfolded conformers to a highly compact native state via an on-pathway intermediate. Here, we investigate the unfolded, intermediate, and native states populated during folding using diffusion single-pair fluorescence resonance energy transfer by measuring the efficiency of energy transfer (or proximity or P ratio) between pairs of fluorophores introduced into the side chains of cysteine residues placed in the center of helices 1 and 4, 1 and 3, or 2 and 4. We show that while the native states of each variant give rise to a single narrow distribution with high P values, the distributions of the intermediates trapped at equilibrium (denoted Ieqm) are fitted by two Gaussian distributions. Modulation of the folding conditions from those that stabilize the intermediate to those that destabilize the intermediate enabled the distribution of lower P value to be assigned to the population of the unfolded ensemble in equilibrium with the intermediate state. The reduced stability of the Ieqm variants allowed analysis of the effect of denaturant concentration on the compaction and breadth of the unfolded state ensemble to be quantified from 0 to 6 M urea. Significant compaction is observed as the concentration of urea is decreased in both the presence and absence of sodium sulfate, as previously reported for a variety of proteins. In the presence of Na2SO4 in 0 M urea, the P value of the unfolded state ensemble approaches that of the native state. Concurrent with compaction, the ensemble displays increased peak width of P values, possibly reflecting a reduction in the rate of conformational exchange among iso-energetic unfolded, but compact conformations. The results provide new insights into the initial stages of folding of Im7 and suggest that the unfolded state is highly conformationally constrained at the outset of folding. PMID:20211187

Single-Molecule Mechanical (Un)folding of RNA Hairpins: Effects of Single A-U to A∙C Pair Substitutions and Single Proton Binding and Implications for mRNA Structure-Induced -1 Ribosomal Frameshifting.

PubMed

Yang, Lixia; Zhong, Zhensheng; Tong, Cailing; Jia, Huan; Liu, Yiran; Chen, Gang

2018-06-08

A wobble A∙C pair can be protonated at near physiological pH to form a more stable wobble A+∙C pair. Here, we constructed an RNA hairpin (rHP) and three mutants with one A-U base pair substituted with an A∙C mismatch on the top (near the loop, U22C), middle (U25C) and bottom (U29C) positions of the stem, respectively. Our results on single-molecule mechanical (un)folding using optical tweezers reveal the destabilization effect of A-U to A∙C pair substitution, and protonation-dependent enhancement of mechanical stability facilitated through an increased folding rate, or decreased unfolding rate, or both. Our data show that protonation may occur rapidly upon the formation of apparent mechanical folding transition state. Furthermore, we measured the bulk -1 ribosomal frameshifting efficiencies of the hairpins by a cell-free translation assay. For the mRNA hairpins studied, -1 frameshifting efficiency correlates with mechanical unfolding force at equilibrium and folding rate at around 15 pN. U29C has a frameshifting efficiency similar to that of rHP (~2%). Accordingly, the bottom 2-4 base pairs of U29C may not form under a stretching force at pH 7.3, which is consistent with the fact that the bottom base pairs of the hairpins may be disrupted by ribosome at the slippery site. U22C and U25C have a similar frameshifting efficiency (~1%), indicating that both unfolding and folding rates of an mRNA hairpin in a crowded environment may affect frameshifting. Our data indicate that mechanical (un)folding of RNA hairpins may mimic how mRNAs unfold and fold in the presence of translating ribosomes.

UFO (UnFold Operator) computer program abstract

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

Kissel, L.; Biggs, F.

UFO (UnFold Operator) is an interactive user-oriented computer program designed to solve a wide range of problems commonly encountered in physical measurements. This document provides a summary of the capabilities of version 3A of UFO.

Beta-Barrel Scaffold of Fluorescent Proteins: Folding, Stability and Role in Chromophore Formation

PubMed Central

Stepanenko, Olesya V.; Stepanenko, Olga V.; Kuznetsova, Irina M.; Verkhusha, Vladislav V.; Turoverov, Konstantin K.

2013-01-01

This review focuses on the current view of the interaction between the β-barrel scaffold of fluorescent proteins and their unique chromophore located in the internal helix. The chromophore originates from the polypeptide chain and its properties are influenced by the surrounding protein matrix of the β-barrel. On the other hand, it appears that a chromophore tightens the β-barrel scaffold and plays a crucial role in its stability. Furthermore, the presence of a mature chromophore causes hysteresis of protein unfolding and refolding. We survey studies measuring protein unfolding and refolding using traditional methods as well as new approaches, such as mechanical unfolding and reassembly of truncated fluorescent proteins. We also analyze models of fluorescent protein unfolding and refolding obtained through different approaches, and compare the results of protein folding in vitro to co-translational folding of a newly synthesized polypeptide chain. PMID:23351712

The Unfolding and Refolding Reactions of Triosephosphate Isomerase from Trypanosoma Cruzi Follow Similar Pathways. Guanidinium Hydrochloride Studies

NASA Astrophysics Data System (ADS)

Vázquez-Contreras, Edgar; Pérez Hernández, Gerardo; Sánchez-Rebollar, Brenda Guadalupe; Chánez-Cárdenas, María Elena

2005-04-01

The unfolding and refolding reactions of Trypanosoma cruzi triosephosphate isomerase (TcTIM) was studied under equilibrium conditions at increasing guanidinium hydrochloride concentrations. The changes in activity intrinsic fluorescence and far-ultraviolet circular dichroism as a function of denaturant were used as a quaternary, tertiary and secondary structural probes respectively. The change in extrinsic ANS fluorescence intensity was also investigated. The results show that the transition between the homodimeric native enzyme to the unfolded monomers (unfolding), and its inverse reaction (refolding) are described by similar pathways and two equilibrium intermediates were detected in both reactions. The mild denaturant concentrations intermediate is active and contains significant amount of secondary and tertiary structures. The medium denaturant concentrations intermediate is inactive and able to bind the fluorescent dye. This intermediates are maybe related with those observed in the denaturation pattern of TIMs from other species; the results are discussed in this context.

Contribution of Long-Range Interactions to the Secondary Structure of an Unfolded Globin

PubMed Central

Fedyukina, Daria V.; Rajagopalan, Senapathy; Sekhar, Ashok; Fulmer, Eric C.; Eun, Ye-Jin; Cavagnero, Silvia

2010-01-01

This work explores the effect of long-range tertiary contacts on the distribution of residual secondary structure in the unfolded state of an α-helical protein. N-terminal fragments of increasing length, in conjunction with multidimensional nuclear magnetic resonance, were employed. A protein representative of the ubiquitous globin fold was chosen as the model system. We found that, while most of the detectable α-helical population in the unfolded ensemble does not depend on the presence of the C-terminal region (corresponding to the native G and H helices), specific N-to-C long-range contacts between the H and A-B-C regions enhance the helical secondary structure content of the N terminus (A-B-C regions). The simple approach introduced here, based on the evaluation of N-terminal polypeptide fragments of increasing length, is of general applicability to identify the influence of long-range interactions in unfolded proteins. PMID:20816043

Dynamical properties of α-amylase in the folded and unfolded state: the role of thermal equilibrium fluctuations for conformational entropy and protein stabilisation

NASA Astrophysics Data System (ADS)

Fitter, J.; Herrmann, R.; Hauß, T.; Lechner, R. E.; Dencher, N. A.

2001-07-01

A comparative analysis of thermal equilibrium fluctuations occurring in a mesophilic and in a thermophilic α-amylase was performed to study the effect of structural fluctuations on thermostability. The thermal fluctuations determining the conformational entropy of both enzymes have been characterised for the folded (at 30°C and 60°C) and for the unfolded state by applying neutron spectroscopy (at 30°C). The folded state shows a higher structural flexibility for the thermophilic protein as compared to the mesophilic homologue. In contrast, the unfolded state of both enzymes is rather similar with respect to the structural fluctuations. On the basis of this result, a mechanism characterised by entropic stabilisation (i.e., smaller Δ S for the unfolding transition of thermophilic α-amylase) can be assumed to be responsible for the higher thermostability of the thermophilic enzyme.

Theoretical and Experimental Investigation of the Translational Diffusion of Proteins in the Vicinity of Temperature-Induced Unfolding Transition.

PubMed

Molchanov, Stanislav; Faizullin, Dzhigangir A; Nesmelova, Irina V

2016-10-06

Translational diffusion is the most fundamental form of transport in chemical and biological systems. The diffusion coefficient is highly sensitive to changes in the size of the diffusing species; hence, it provides important information on the variety of macromolecular processes, such as self-assembly or folding-unfolding. Here, we investigate the behavior of the diffusion coefficient of a macromolecule in the vicinity of heat-induced transition from folded to unfolded state. We derive the equation that describes the diffusion coefficient of the macromolecule in the vicinity of the transition and use it to fit the experimental data from pulsed-field-gradient nuclear magnetic resonance (PFG NMR) experiments acquired for two globular proteins, lysozyme and RNase A, undergoing temperature-induced unfolding. A very good qualitative agreement between the theoretically derived diffusion coefficient and experimental data is observed.

Analyzing phenological extreme events over the past five decades in Germany

NASA Astrophysics Data System (ADS)

Schleip, Christoph; Menzel, Annette; Estrella, Nicole; Graeser, Philipp

2010-05-01

As climate change may alter the frequency and intensity of extreme temperatures, we analysed whether warming of the last 5 decades has already changed the statistics of phenological extreme events. In this context, two extreme value statistical concepts are discussed and applied to existing phenological datasets of German Weather Service (DWD) in order to derive probabilities of occurrence for extreme early or late phenological events. We analyse four phenological groups; "begin of flowering, "leaf foliation", "fruit ripening" and "leaf colouring" as well as DWD indicator phases of the "phenological year". Additionally we put an emphasis on a between-species analysis; a comparison of differences in extreme onsets between three common northern conifers. Furthermore we conducted a within-species analysis with different phases of horse chestnut throughout a year. The first statistical approach fits data to a Gaussian model using traditional statistical techniques, and then analyses the extreme quantile. The key point of this approach is the adoption of an appropriate probability density function (PDF) to the observed data and the assessment of the PDF parameters change in time. The full analytical description in terms of the estimated PDF for defined time steps of the observation period allows probability assessments of extreme values for e.g. annual or decadal time steps. Related with this approach is the possibility of counting out the onsets which fall in our defined extreme percentiles. The estimation of the probability of extreme events on the basis of the whole data set is in contrast to analyses with the generalized extreme value distribution (GEV). The second approach deals with the extreme PDFs itself and fits the GEV distribution to annual minima of phenological series to provide useful estimates about return levels. For flowering and leaf unfolding phases exceptionally early extremes are seen since the mid 1980s and especially for the single years 1961, 1990 and 2007 whereas exceptionally extreme late events are seen in the year 1970. Summer phases such as fruit ripening exhibit stronger shifts to early extremes than spring phases. Leaf colouring phases reveal increasing probability for late extremes. The with GEV estimated 100-year event of Picea, Pinus and Larix amount to extreme early events of about -27, -31.48 and -32.79 days, respectively. If we assume non-stationary minimum data we get a more extreme 100-year event of about -35.40 for Picea but associated with wider confidence intervals. The GEV is simply another probability distribution but for purposes of extreme analysis in phenology it should be considered as equally important as (if not more important than) the Gaussian PDF approach.

Microsecond simulations of the folding/unfolding thermodynamics of the Trp-cage mini protein

PubMed Central

Day, Ryan; Paschek, Dietmar; Garcia, Angel E.

2012-01-01

We study the unbiased folding/unfolding thermodynamics of the Trp-cage miniprotein using detailed molecular dynamics simulations of an all-atom model of the protein in explicit solvent, using the Amberff99SB force field. Replica-exchange molecular dynamics (REMD) simulations are used to sample the protein ensembles over a broad range of temperatures covering the folded and unfolded states, and at two densities. The obtained ensembles are shown to reach equilibrium in the 1 μs per replica timescale. The total simulation time employed in the calculations exceeds 100 μs. Ensemble averages of the fraction folded, pressure, and energy differences between the folded and unfolded states as a function of temperature are used to model the free energy of the folding transition, ΔG(P,T), over the whole region of temperature and pressures sampled in the simulations. The ΔG(P,T) diagram describes an ellipse over the range of temperatures and pressures sampled, predicting that the system can undergo pressure induced unfolding and cold denaturation at low temperatures and high pressures, and unfolding at low pressures and high temperatures. The calculated free energy function exhibits remarkably good agreement with the experimental folding transition temperature (Tf = 321 K), free energy and specific heat changes. However, changes in enthalpy and entropy are significantly different than the experimental values. We speculate that these differences may be due to the simplicity of the semi-empirical force field used in the simulations and that more elaborate force fields may be required to describe appropriately the thermodynamics of proteins. PMID:20408169

Equilibrium and kinetic folding of rabbit muscle triosephosphate isomerase by hydrogen exchange mass spectrometry.

PubMed

Pan, Hai; Raza, Ashraf S; Smith, David L

2004-03-05

Unfolding and refolding of rabbit muscle triosephosphate isomerase (TIM), a model for (betaalpha)8-barrel proteins, has been studied by amide hydrogen exchange/mass spectrometry. Unfolding was studied by destabilizing the protein in guanidine hydrochloride (GdHCl) or urea, pulse-labeling with 2H2O and analyzing the intact protein by HPLC electrospray ionization mass spectrometry. Bimodal isotope patterns were found in the mass spectra of the labeled protein, indicating two-state unfolding behavior. Refolding experiments were performed by diluting solutions of TIM unfolded in GdHCl or urea and pulse-labeling with 2H2O at different times. Mass spectra of the intact protein labeled after one to two minutes had three envelopes of isotope peaks, indicating population of an intermediate. Kinetic modeling indicates that the stability of the folding intermediate in water is only 1.5 kcal/mol. Failure to detect the intermediate in the unfolding experiments was attributed to its low stability and the high concentrations of denaturant required for unfolding experiments. The folding status of each segment of the polypeptide backbone was determined from the deuterium levels found in peptic fragments of the labeled protein. Analysis of these spectra showed that the C-terminal half folds to form the intermediate, which then forms native TIM with folding of the N-terminal half. These results show that TIM folding fits the (4+4) model for folding of (betaalpha)8-barrel proteins. Results of a double-jump experiment indicate that proline isomerization does not contribute to the rate-limiting step in the folding of TIM.

An alternatively spliced heat shock transcription factor, OsHSFA2dI, functions in the heat stress-induced unfolded protein response in rice.

PubMed

Cheng, Q; Zhou, Y; Liu, Z; Zhang, L; Song, G; Guo, Z; Wang, W; Qu, X; Zhu, Y; Yang, D

2015-03-01

As sessile organisms, plants have evolved a wide range of defence pathways to cope with environmental stress such as heat shock. However, the molecular mechanism of these defence pathways remains unclear in rice. In this study, we found that OsHSFA2d, a heat shock transcriptional factor, encodes two main splice variant proteins, OsHSFA2dI and OsHSFA2dII in rice. Under normal conditions, OsHSFA2dII is the dominant but transcriptionally inactive spliced form. However, when the plant suffers heat stress, OsHSFA2d is alternatively spliced into a transcriptionally active form, OsHSFA2dI, which participates in the heat stress response (HSR). Further study found that this alternative splicing was induced by heat shock rather than photoperiod. We found that OsHSFA2dI is localised to the nucleus, whereas OsHSFA2dII is localised to the nucleus and cytoplasm. Moreover, expression of the unfolded protein response (UNFOLDED PROTEIN RESPONSE) sensors, OsIRE1, OsbZIP39/OsbZIP60 and the UNFOLDED PROTEIN RESPONSE marker OsBiP1, was up-regulated. Interestingly, OsbZIP50 was also alternatively spliced under heat stress, indicating that UNFOLDED PROTEIN RESPONSE signalling pathways were activated by heat stress to re-establish cellular protein homeostasis. We further demonstrated that OsHSFA2dI participated in the unfolded protein response by regulating expression of OsBiP1. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

Engineered Human Antibody Constant Domains with Increased Stability*S⃞

PubMed Central

Gong, Rui; Vu, Bang K.; Feng, Yang; Prieto, DaRue A.; Dyba, Marzena A.; Walsh, Joseph D.; Prabakaran, Ponraj; Veenstra, Timothy D.; Tarasov, Sergey G.; Ishima, Rieko; Dimitrov, Dimiter S.

2009-01-01

The immunoglobulin (Ig) constant CH2 domain is critical for antibody effector functions. Isolated CH2 domains are promising as scaffolds for construction of libraries containing diverse binders that could also confer some effector functions. However, previous work has shown that an isolated murine CH2 domain is relatively unstable to thermally induced unfolding. To explore unfolding mechanisms of isolated human CH2 and increase its stability γ1 CH2 was cloned and a panel of cysteine mutants was constructed. Human γ1 CH2 unfolded at a higher temperature (Tm = 54.1 °C, as measured by circular dichroism) than that previously reported for a mouse CH2 (41 °C). One mutant (m01) was remarkably stable (Tm = 73.8 °C). Similar results were obtained by differential scanning calorimetry. This mutant was also significantly more stable than the wild-type CH2 against urea induced unfolding (50% unfolding at urea concentration of 6.8 m versus 4.2 m). The m01 was highly soluble and monomeric. The existence of the second disulfide bond in m01 and its correct position were demonstrated by mass spectrometry and nuclear magnetic resonance spectroscopy, respectively. The loops were on average more flexible than the framework in both CH2 and m01, and the overall secondary structure was not affected by the additional disulfide bond. These data suggest that a human CH2 domain is relatively stable to unfolding at physiological temperature, and that both CH2 and the highly stable mutant m01 are promising new scaffolds for the development of therapeutics against human diseases. PMID:19307178

Force-dependent switch in protein unfolding pathways and transition-state movements

PubMed Central

Zhuravlev, Pavel I.; Hinczewski, Michael; Chakrabarti, Shaon; Marqusee, Susan; Thirumalai, D.

2016-01-01

Although it is known that single-domain proteins fold and unfold by parallel pathways, demonstration of this expectation has been difficult to establish in experiments. Unfolding rate, ku(f), as a function of force f, obtained in single-molecule pulling experiments on src SH3 domain, exhibits upward curvature on a log⁡ku(f) plot. Similar observations were reported for other proteins for the unfolding rate ku([C]). These findings imply unfolding in these single-domain proteins involves a switch in the pathway as f or [C] is increased from a low to a high value. We provide a unified theory demonstrating that if log⁡ku as a function of a perturbation (f or [C]) exhibits upward curvature then the underlying energy landscape must be strongly multidimensional. Using molecular simulations we provide a structural basis for the switch in the pathways and dramatic shifts in the transition-state ensemble (TSE) in src SH3 domain as f is increased. We show that a single-point mutation shifts the upward curvature in log⁡ku(f) to a lower force, thus establishing the malleability of the underlying folding landscape. Our theory, applicable to any perturbation that affects the free energy of the protein linearly, readily explains movement in the TSE in a β-sandwich (I27) protein and single-chain monellin as the denaturant concentration is varied. We predict that in the force range accessible in laser optical tweezer experiments there should be a switch in the unfolding pathways in I27 or its mutants. PMID:26818842

Integrated view of internal friction in unfolded proteins from single-molecule FRET, contact quenching, theory, and simulations

PubMed Central

Soranno, Andrea; Holla, Andrea; Dingfelder, Fabian; Nettels, Daniel; Makarov, Dmitrii E.; Schuler, Benjamin

2017-01-01

Internal friction is an important contribution to protein dynamics at all stages along the folding reaction. Even in unfolded and intrinsically disordered proteins, internal friction has a large influence, as demonstrated with several experimental techniques and in simulations. However, these methods probe different facets of internal friction and have been applied to disparate molecular systems, raising questions regarding the compatibility of the results. To obtain an integrated view, we apply here the combination of two complementary experimental techniques, simulations, and theory to the same system: unfolded protein L. We use single-molecule Förster resonance energy transfer (FRET) to measure the global reconfiguration dynamics of the chain, and photoinduced electron transfer (PET), a contact-based method, to quantify the rate of loop formation between two residues. This combination enables us to probe unfolded-state dynamics on different length scales, corresponding to different parts of the intramolecular distance distribution. Both FRET and PET measurements show that internal friction dominates unfolded-state dynamics at low denaturant concentration, and the results are in remarkable agreement with recent large-scale molecular dynamics simulations using a new water model. The simulations indicate that intrachain interactions and dihedral angle rotation correlate with the presence of internal friction, and theoretical models of polymer dynamics provide a framework for interrelating the contribution of internal friction observed in the two types of experiments and in the simulations. The combined results thus provide a coherent and quantitative picture of internal friction in unfolded proteins that could not be attained from the individual techniques. PMID:28223518

Integrated view of internal friction in unfolded proteins from single-molecule FRET, contact quenching, theory, and simulations.

PubMed

Soranno, Andrea; Holla, Andrea; Dingfelder, Fabian; Nettels, Daniel; Makarov, Dmitrii E; Schuler, Benjamin

2017-03-07

Internal friction is an important contribution to protein dynamics at all stages along the folding reaction. Even in unfolded and intrinsically disordered proteins, internal friction has a large influence, as demonstrated with several experimental techniques and in simulations. However, these methods probe different facets of internal friction and have been applied to disparate molecular systems, raising questions regarding the compatibility of the results. To obtain an integrated view, we apply here the combination of two complementary experimental techniques, simulations, and theory to the same system: unfolded protein L. We use single-molecule Förster resonance energy transfer (FRET) to measure the global reconfiguration dynamics of the chain, and photoinduced electron transfer (PET), a contact-based method, to quantify the rate of loop formation between two residues. This combination enables us to probe unfolded-state dynamics on different length scales, corresponding to different parts of the intramolecular distance distribution. Both FRET and PET measurements show that internal friction dominates unfolded-state dynamics at low denaturant concentration, and the results are in remarkable agreement with recent large-scale molecular dynamics simulations using a new water model. The simulations indicate that intrachain interactions and dihedral angle rotation correlate with the presence of internal friction, and theoretical models of polymer dynamics provide a framework for interrelating the contribution of internal friction observed in the two types of experiments and in the simulations. The combined results thus provide a coherent and quantitative picture of internal friction in unfolded proteins that could not be attained from the individual techniques.

The secondary structure and the thermal unfolding parameters of the S-layer protein from Lactobacillus salivarius.

PubMed

Lighezan, Liliana; Georgieva, Ralitsa; Neagu, Adrian

2016-09-01

Surface layer (S-layer) proteins have been identified in the cell envelope of many organisms, such as bacteria and archaea. They self-assemble, forming monomolecular crystalline arrays. Isolated S-layer proteins are able to recrystallize into regular lattices, which proved useful in biotechnology. Here we investigate the structure and thermal unfolding of the S-layer protein isolated from Lactobacillus salivarius 16 strain of human origin. Using circular dichroism (CD) spectroscopy, and the software CDSSTR from DICHROWEB, CONTINLL from CDPro, as well as CDNN, we assess the fractions of the protein's secondary structural elements at temperatures ranging between 10 and 90 °C, and predict the tertiary class of the protein. To study the thermal unfolding of the protein, we analyze the temperature dependence of the CD signal in the far- and near-UV domains. Fitting the experimental data by two- and three-state models of thermal unfolding, we infer the midpoint temperatures, the temperature dependence of the changes in Gibbs free energy, enthalpy, and entropy of the unfolding transitions in standard conditions, and the temperature dependence of the equilibrium constant. We also estimate the changes in heat capacity at constant pressure in standard conditions. The results indicate that the thermal unfolding of the S-layer protein from L. salivarius is highly cooperative, since changes in the secondary and tertiary structures occur simultaneously. The thermodynamic analysis predicts a "cold" transition, at about -3 °C, of both the secondary and tertiary structures. Our findings may be important for the use of S-layer proteins in biotechnology and in biomedical applications.

Force-dependent switch in protein unfolding pathways and transition-state movements.

PubMed

Zhuravlev, Pavel I; Hinczewski, Michael; Chakrabarti, Shaon; Marqusee, Susan; Thirumalai, D

2016-02-09

Although it is known that single-domain proteins fold and unfold by parallel pathways, demonstration of this expectation has been difficult to establish in experiments. Unfolding rate, [Formula: see text], as a function of force f, obtained in single-molecule pulling experiments on src SH3 domain, exhibits upward curvature on a [Formula: see text] plot. Similar observations were reported for other proteins for the unfolding rate [Formula: see text]. These findings imply unfolding in these single-domain proteins involves a switch in the pathway as f or [Formula: see text] is increased from a low to a high value. We provide a unified theory demonstrating that if [Formula: see text] as a function of a perturbation (f or [Formula: see text]) exhibits upward curvature then the underlying energy landscape must be strongly multidimensional. Using molecular simulations we provide a structural basis for the switch in the pathways and dramatic shifts in the transition-state ensemble (TSE) in src SH3 domain as f is increased. We show that a single-point mutation shifts the upward curvature in [Formula: see text] to a lower force, thus establishing the malleability of the underlying folding landscape. Our theory, applicable to any perturbation that affects the free energy of the protein linearly, readily explains movement in the TSE in a β-sandwich (I27) protein and single-chain monellin as the denaturant concentration is varied. We predict that in the force range accessible in laser optical tweezer experiments there should be a switch in the unfolding pathways in I27 or its mutants.

[Effects of cropping patterns on photosynthesis characteristics of summer maize and its utilization of solar and heat resources].

PubMed

Zhu, Yuan-Gang; Dong, Shu-Ting; Zhang, Ji-Wang; Liu, Peng; Yang, Jin-Sheng; Jia, Chun-Lan; Liu, Jing-Guo; Li, Deng-Hai

2010-06-01

In order to investigate the effects of interplanting and direct seeding on the photosynthesis characteristics of summer maize and its utilization of solar and heat resources, two summer maize cultivars (Zhengdan 958 and Denghai 661) were planted in the farmlands of Denghai Seed Co. Ltd in Laizhou City of Shandong Province, with 67500 plants x hm(-2) and three sowing dates. The above-ground biomass, plant growth rate, leaf area index, and net photosynthetic rate per ear leaf were measured to reveal the photosynthesis characteristics of test cultivars. In the meantime, the characters of grain-filling were simulated by Richards' model, and the solar resource utilization efficiency of the cultivars was calculated, in combining with meteorological data. Comparing with interplanting, direct seeding increased the grain yield by 1.17%-3.33%, but decreased the thousand-grain weight significantly. Growth stages were extended under earlier sowing. The leaf area index and net photosynthetic rate from flowering to 30 d after anthesis were significantly higher under direct seeding than under interplanting, but after then, they decreased faster. Direct seeding induced a higher accumulation of dry matter and a faster plant growth rate before and after flowering. Under direct seeding, the maximum grain-filling rate reached earlier, the starting potential was higher, but the grain-filling period, active grain-filling period, and W(max) were lower, compared with those under interplanting. Also under direct seeding, the total accumulative temperature and solar radiation during growth period decreased by 150-350 degrees C x d and 200-400 MJ x m(-2), respectively, but the solar resource utilization efficiency of grain increased by 10.5%-24.7%. All the results suggested that direct seeding was superior to interplanting for the summer maize production under field condition. In order to enhance solar and heat utilization efficiency and excavate yield potential, it would be essential to improve the leaf photosynthesis efficiency and postpone leaf aging.

Effects of oil on internal gas transport, radial oxygen loss, gas films and bud growth in Phragmites australis.

PubMed

Armstrong, Jean; Keep, Rory; Armstrong, William

2009-01-01

Oil pollution of wetlands is a world-wide problem but, to date, research has concentrated on its influences on salt marsh rather than freshwater plant communities. The effects of water-borne light oils (liquid paraffin and diesel) were investigated on the fresh/brackish wetland species Phragmites australis in terms of routes of oil infiltration, internal gas transport, radial O(2) loss (ROL), underwater gas films and bud growth. Pressure flow resistances of pith cavities of nodes and aerenchyma of leaf sheaths, with or without previous exposure to oil, were recorded from flow rates under applied pressure. Convective flows were measured from living excised culms with oiled and non-oiled nodes and leaf sheaths. The effect of oil around culm basal nodes on ROL from rhizome and root apices was measured polarographically. Surface gas films on submerged shoots with and without oil treatment were recorded photographically. Growth and emergence of buds through water with and without an oil film were measured. Internodes are virtually impermeable, but nodes of senesced and living culms are permeable to oils which can block pith cavity diaphragms, preventing flows at applied pressures of 1 kPa, natural convective transport to the rhizome, and greatly decreasing ROL to phyllospheres and rhizospheres. Oil infiltrating or covering living leaf sheaths prevents humidity-induced convection. Oil displaces surface gas films from laminae and leaf sheaths. Buds emerge only a few centimetres through oil and die. Oil infiltrates the gas space system via nodal and leaf sheath stomata, reducing O(2) diffusion and convective flows into the rhizome system and decreasing oxygenation of phyllospheres and rhizospheres; underwater gas exchange via gas films will be impeded. Plants can be weakened by oil-induced failure of emerging buds. Plants will be most at risk during the growing season.

Thrips (Thysanoptera: Thripidae) mitigation in seedling cotton using strip tillage and winter cover crops.

PubMed

Toews, Michael D; Tubbs, R Scott; Wann, Dylan Q; Sullivan, Dana

2010-10-01

Thrips are the most consistent insect pests of seedling cotton in the southeastern United States, where symptoms can range from leaf curling to stand loss. In a 2 year study, thrips adults and immatures were sampled at 14, 21 and 28 days after planting on cotton planted with a thiamethoxam seed treatment in concert with crimson clover, wheat or rye winter cover crops and conventional or strip tillage to investigate potential differences in thrips infestations. Densities of adult thrips, primarily Frankliniella fusca (Hinds), peaked on the first sampling date, whereas immature densities peaked on the second sampling date. Regardless of winter cover crop, plots that received strip tillage experienced significantly fewer thrips at each sampling interval. In addition, assessment of percentage ground cover 42 days after planting showed that there was more than twice as much ground cover in the strip-tilled plots compared with conventionally tilled plots. Correlation analyses showed that increased ground cover was inversely related to thrips densities that occurred on all three sampling dates in 2008 and the final sampling date in 2009. Growers who utilize strip tillage and a winter cover crop can utilize seed treatments for mitigation of early-season thrips infestation.

Improvement of In Vitro Date Palm Plantlet Acclimatization Rate with Kinetin and Hoagland Solution.

PubMed

Hassan, Mona M

2017-01-01

In vitro propagation of date palm Phoenix dactylifera L. is an ideal method to produce large numbers of healthy plants with specific characteristics and has the ability to transfer plantlets to ex vitro conditions at low cost and with a high survival rate. This chapter describes optimized acclimatization procedures for in vitro date palm plantlets. Primarily, the protocol presents the use of kinetin and Hoagland solution to enhance the growth of Barhee cv. plantlets in the greenhouse at two stages of acclimatization and the appropriate planting medium under shade and sunlight in the nursery. Foliar application of kinetin (20 mg/L) is recommended at the first stage. A combination between soil and foliar application of 50% Hoagland solution is favorable to plant growth and developmental parameters including plant height, leaf width, stem base diameter, chlorophyll A and B, carotenoids, and indoles. The optimum values of vegetative growth parameters during the adaptation stage in a shaded nursery are achieved using planting medium containing peat moss/perlite 2:1 (v/v), while in a sunlight nursery, clay/perlite/compost at equal ratio is the best. This protocol is suitable for large-scale production of micropropagated date palm plantlets.

Kinetic evidence for folding and unfolding intermediates in staphylococcal nuclease.

PubMed

Walkenhorst, W F; Green, S M; Roder, H

1997-05-13

The complex kinetic behavior commonly observed in protein folding studies suggests that a heterogeneous population of molecules exists in solution and that a number of discrete steps are involved in the conversion of unfolded molecules to the fully native form. A central issue in protein folding is whether any of these kinetic events represent conformational steps important for efficient folding rather than side reactions caused by slow steps such as proline isomerization or misfolding of the polypeptide chain. In order to address this question, we used stopped-flow fluorescence techniques to characterize the kinetic mechanism of folding and unfolding for a Pro- variant of SNase in which all six proline residues were replaced by glycines or alanines. Compared to the wild-type protein, which exhibits a series of proline-dependent slow folding phases, the folding kinetics of Pro- SNase were much simpler, which made quantitative kinetic analysis possible. Despite the absence of prolines or other complicating factors, the folding kinetics still contain several phases and exhibit a complex denaturant dependence. The GuHCl dependence of the major observable folding phase and a distinct lag in the appearance of the native state provide clear evidence for an early folding intermediate. The fluorescence of Trp140 in the alpha-helical domain is insensitive to the formation of this early intermediate, which is consistent with a partially folded state with a stable beta-domain and a largely disordered alpha-helical region. A second intermediate is required to model the kinetics of unfolding for the Pro- variant, which shows evidence for a denaturant-induced change in the rate-limiting unfolding step. With the inclusion of these two intermediates, we are able to completely model the major phase(s) in both folding and unfolding across a wide range of denaturant concentrations using a sequential four-state folding mechanism. In order to model the minor slow phase observed for the Pro- mutant, a six-state scheme containing a parallel pathway originating from a distinct unfolded state was required. The properties of this alternate unfolded conformation are consistent with those expected due to the presence of a non-prolyl cis peptide bond. To test the kinetic model, we used simulations based on the six-state scheme and were able to completely reproduce the folding kinetics for Pro- SNase across a range of denaturant concentrations.

Prediction and analysis of structure, stability and unfolding of thermolysin-like proteases

NASA Astrophysics Data System (ADS)

Vriend, Gert; Eijsink, Vincent

1993-08-01

Bacillus neutral proteases (NPs) form a group of well-characterized homologous enzymes, that exhibit large differences in thermostability. The three-dimensional (3D) structures of several of these enzymes have been modelled on the basis of the crystal structures of the NPs of B. thermoproteolyticus (thermolysin) and B. cercus. Several new techniques have been developed to improve the model-building procedures. Also a model-building by mutagenesis' strategy was used, in which mutants were designed just to shed light on parts of the structures that were particularly hard to model. The NP models have been used for the prediction of site-directed mutations aimed at improving the thermostability of the enzymes. Predictions were made using several novel computational techniques, such as position-specific rotamer searching, packing quality analysis and property-profile database searches. Many stabilizing mutations were predicted and produced: improvement of hydrogen bonding, exclusion of buried water molecules, capping helices, improvement of hydrophobic interactions and entropic stabilization have been applied successfully. At elevated temperatures NPs are irreversibly inactivated as a result of autolysis. It has been shown that this denaturation process is independent of the protease activity and concentration and that the inactivation follows first-order kinetics. From this it has been conjectured that local unfolding of (surface) loops, which renders the protein susceptible to autolysis, is the rate-limiting step. Despite the particular nature of the thermal denaturation process, normal rules for protein stability can be applied to NPs. However, rather than stabilizing the whole protein against global unfolding, only a small region has to be protected against local unfolding. In contrast to proteins in general, mutational effects in proteases are not additive and their magnitude is strongly dependent on the location of the mutation. Mutations that alter the stability of the NP by a large amount are located in a relatively weak region (or more precisely, they affect a local unfolding pathway with a relatively low free energy of activation). One weak region, that is supposedly important in the early steps of NP unfolding, has been determined in the NP of B. stearothermophilus. After eliminating this weakest link a drastic increase in thermostability was observed and the search for the second-weakest link, or the second-lowest energy local unfolding pathway is now in progress. Hopefully, this approach can be used to unravel the entire early phase of unfolding.

How Adequate are One- and Two-Dimensional Free Energy Landscapes for Protein Folding Dynamics?

NASA Astrophysics Data System (ADS)

Maisuradze, Gia G.; Liwo, Adam; Scheraga, Harold A.

2009-06-01

The molecular dynamics trajectories of protein folding or unfolding, generated with the coarse-grained united-residue force field for the B domain of staphylococcal protein A, were analyzed by principal component analysis (PCA). The folding or unfolding process was examined by using free-energy landscapes (FELs) in PC space. By introducing a novel multidimensional FEL, it was shown that the low-dimensional FELs are not always sufficient for the description of folding or unfolding processes. Similarities between the topographies of FELs along low- and high-indexed principal components were observed.

Single molecule FRET investigation of pressure-driven unfolding of cold shock protein A

NASA Astrophysics Data System (ADS)

Schneider, Sven; Paulsen, Hauke; Reiter, Kim Colin; Hinze, Erik; Schiene-Fischer, Cordelia; Hübner, Christian G.

2018-03-01

We demonstrate that fused silica capillaries are suitable for single molecule fluorescence resonance energy transfer (smFRET) measurements at high pressure with an optical quality comparable to the measurement on microscope coverslips. Therefore, we optimized the imaging conditions in a standard square fused silica capillary with an adapted arrangement and evaluated the performance by imaging the focal volume, fluorescence correlation spectroscopy benchmarks, and FRET measurements. We demonstrate single molecule FRET measurements of cold shock protein A unfolding at a pressure up to 2000 bars and show that the unfolded state exhibits an expansion almost independent of pressure.

A fluorescence approach to the unfolding thermodynamics of horseradish peroxidase based on heme degradation by hydrogen peroxide

NASA Astrophysics Data System (ADS)

Ke, Zhigang; Ma, Shanshan; Li, Lamei; Huang, Qing

2016-07-01

Horseradish peroxidase (HRP) is a classical heme-containing protein which has been applied in many fields. The prosthetic group heme in HRP, especially in unfolded state, can react with hydrogen peroxide (H2O2) to produce a fluorescent product with the maximum emission wavelength at 450 nm. Utilizing this emission band as a fluorescence probe, the unfolding process of HRP in urea can be assessed quantitatively, and the calculated thermodynamic parameters are consistent with those determined by circular dichroism (CD) at 222 nm and steady-state tryptophan (Trp) fluorescence methods.

Role of electrostatic interaction on surfactant induced protein unfolding

NASA Astrophysics Data System (ADS)

Sumit, Kumar, Sugam; Aswal, V. K.

2013-02-01

Small Angle Neutron Scattering has been used to examine the effect of electrostatic interaction on surfactant induced protein unfolding. Measurements are carried out from 1 wt% Bovine Serum Albumin (BSA) protein with 1 wt% Sodium Dodecyl Sulphate (SDS) surfactant at pH 7 in presence of varying concentration of NaCl. It is found that both the components (protein and surfactant micelle which are likely charged) exist individually without any interaction in absence of salt, whereas their interaction and protein unfolding is enhanced with the increase in salt concentration. The structure of protein-surfactant interaction is characterized by fractal bead-necklace model.

Breakdown of a 2D Heteroclinic Connection in the Hopf-Zero Singularity (I)

NASA Astrophysics Data System (ADS)

Baldomá, I.; Castejón, O.; Seara, T. M.

2018-04-01

In this paper we study a beyond all orders phenomenon which appears in the analytic unfoldings of the Hopf-zero singularity. It consists in the breakdown of a two-dimensional heteroclinic surface which exists in the truncated normal form of this singularity at any order. The results in this paper are twofold: on the one hand, we give results for generic unfoldings which lead to sharp exponentially small upper bounds of the difference between these manifolds. On the other hand, we provide asymptotic formulas for this difference by means of the Melnikov function for some non-generic unfoldings.

Unfolding and folding internal friction of β-hairpins is smaller than that of α-helices.

PubMed

Schulz, Julius C F; Miettinen, Markus S; Netz, R R

2015-04-02

By the forced unfolding of polyglutamine and polyalanine homopeptides in competing α-helix and β-hairpin secondary structures, we disentangle equilibrium free energetics from nonequilibrium dissipative effects. We find that α-helices are characterized by larger friction or dissipation upon unfolding, regardless of whether they are free energetically preferred over β-hairpins or not. Our analysis, based on MD simulations for atomistic peptide models with explicit water, suggests that this difference is related to the internal friction and mostly caused by the different number of intrapeptide hydrogen bonds in the α-helix and β-hairpin states.

The impact of the unfolded protein response on human disease

PubMed Central

Wang, Shiyu

2012-01-01

A central function of the endoplasmic reticulum (ER) is to coordinate protein biosynthetic and secretory activities in the cell. Alterations in ER homeostasis cause accumulation of misfolded/unfolded proteins in the ER. To maintain ER homeostasis, eukaryotic cells have evolved the unfolded protein response (UPR), an essential adaptive intracellular signaling pathway that responds to metabolic, oxidative stress, and inflammatory response pathways. The UPR has been implicated in a variety of diseases including metabolic disease, neurodegenerative disease, inflammatory disease, and cancer. Signaling components of the UPR are emerging as potential targets for intervention and treatment of human disease. PMID:22733998

Time-dependent, x-ray spectral unfolds and brightness temperatures for intense Li{sup +} ion beam-driven hohlraums

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

Fehl, D.L.; Chandler, G.A.; Biggs, F.

X-ray-producing hohlraums are being studied as indirect drives for inertial confinement fusion targets. In a 1994 target series on the PBFAII accelerator, cylindrical hohlraum targets were heated by an intense Li{sup +} ion beam and viewed by an array of 13 time-resolved, filtered x-ray detectors (XRDs). The unfold operator (UFO) code and its suite of auxiliary functions were used extensively in obtaining time-resolved x-ray spectra and radiation temperatures from this diagnostic. The UFO was also used to obtain fitted response functions from calibration data, to simulate data from blackbody x-ray spectra of interest, to determine the suitability of various unfoldingmore » parameters (e.g., energy domain, energy partition, smoothing conditions, and basis functions), to interpolate the XRD signal traces, and to unfold experimental data. The simulation capabilities of the code were useful in understanding an anomalous feature in the unfolded spectra at low photon energies ({le}100 eV). Uncertainties in the differential and energy-integrated unfolded spectra were estimated from uncertainties in the data. The time{endash}history of the radiation temperature agreed well with independent calculations of the wall temperature in the hohlraum. {copyright} {ital 1997 American Institute of Physics.}« less

Developing a Novel, Interdisciplinary Approach to Study Protein Unfolding

NASA Astrophysics Data System (ADS)

Bentley, Ian; Link, Justin

2013-03-01

The ability of a protein to function is a direct result of its ability to properly obtain its native, folded structure. In order to determine the structural stability of proteins and to gain knowledge of their folding mechanism, we must develop protocols that allow us to monitor the controlled unfolding of proteins. Here, we investigate the stability of cytochrome c, a well-studied, model protein, under denaturing conditions using circular dichroism (CD) and fluorescence. Using either a chemical denaturant (Guanidine HCl) or heat, we can cause a protein to gradually unfold. The changes in the fluorescence and CD spectra can provide insight into the stability of proteins by providing us with thermodynamic parameters such as the Gibbs free energy, melting temperature and enthalpy. Research in this lab has been explored with mutant proteins and change in CD signal, however further work must still be done to observe their unfolding monitored by fluorescence. This technique will allow us to determine which regions of native cytochrome c have the greatest impact on the protein folding process. The objective of this session is to present recent work in developing a protocol to observe the unfolding of wild type and mutant proteins with fluorescence. The Borcer Fund, The John A. Hauck Foundation, and Xavier University

Conformational stability of apoflavodoxin.

PubMed Central

Genzor, C. G.; Beldarraín, A.; Gómez-Moreno, C.; López-Lacomba, J. L.; Cortijo, M.; Sancho, J.

1996-01-01

Flavodoxins are alpha/beta proteins that mediate electron transfer reactions. The conformational stability of apoflavodoxin from Anaboena PCC 7119 has been studied by calorimetry and urea denaturation as a function of pH and ionic strength. At pH > 12, the protein is unfolded. Between pH 11 and pH 6, the apoprotein is folded properly as judged from near-ultraviolet (UV) circular dichroism (CD) and high-field 1H NMR spectra. In this pH interval, apoflavodoxin is a monomer and its unfolding by urea or temperature follows a simple two-state mechanism. The specific heat capacity of unfolding for this native conformation is unusually low. Near its isoelectric point (3.9), the protein is highly insoluble. At lower pH values (pH 3.5-2.0), apoflavodoxin adopts a conformation with the properties of a molten globule. Although apoflavodoxin at pH 2 unfolds cooperatively with urea in a reversible fashion and the fluorescence and far-UV CD unfolding curves coincide, the transition midpoint depends on the concentration of protein, ruling out a simple two-state process at acidic pH. Apoflavodoxin constitutes a promising system for the analysis of the stability and folding of alpha/beta proteins and for the study of the interaction between apoflavoproteins and their corresponding redox cofactors. PMID:8819170

The RtcB RNA ligase is an essential component of the metazoan unfolded protein response.

PubMed

Kosmaczewski, Sara Guckian; Edwards, Tyson James; Han, Sung Min; Eckwahl, Matthew J; Meyer, Benjamin Isaiah; Peach, Sally; Hesselberth, Jay R; Wolin, Sandra L; Hammarlund, Marc

2014-12-01

RNA ligation can regulate RNA function by altering RNA sequence, structure and coding potential. For example, the function of XBP1 in mediating the unfolded protein response requires RNA ligation, as does the maturation of some tRNAs. Here, we describe a novel in vivo model in Caenorhabditis elegans for the conserved RNA ligase RtcB and show that RtcB ligates the xbp-1 mRNA during the IRE-1 branch of the unfolded protein response. Without RtcB, protein stress results in the accumulation of unligated xbp-1 mRNA fragments, defects in the unfolded protein response, and decreased lifespan. RtcB also ligates endogenous pre-tRNA halves, and RtcB mutants have defects in growth and lifespan that can be bypassed by expression of pre-spliced tRNAs. In addition, animals that lack RtcB have defects that are independent of tRNA maturation and the unfolded protein response. Thus, RNA ligation by RtcB is required for the function of multiple endogenous target RNAs including both xbp-1 and tRNAs. RtcB is uniquely capable of performing these ligation functions, and RNA ligation by RtcB mediates multiple essential processes in vivo. © 2014 The Authors.

The RtcB RNA ligase is an essential component of the metazoan unfolded protein response

PubMed Central

Kosmaczewski, Sara Guckian; Edwards, Tyson James; Han, Sung Min; Eckwahl, Matthew J; Meyer, Benjamin Isaiah; Peach, Sally; Hesselberth, Jay R; Wolin, Sandra L; Hammarlund, Marc

2014-01-01

RNA ligation can regulate RNA function by altering RNA sequence, structure and coding potential. For example, the function of XBP1 in mediating the unfolded protein response requires RNA ligation, as does the maturation of some tRNAs. Here, we describe a novel in vivo model in Caenorhabditis elegans for the conserved RNA ligase RtcB and show that RtcB ligates the xbp-1 mRNA during the IRE-1 branch of the unfolded protein response. Without RtcB, protein stress results in the accumulation of unligated xbp-1 mRNA fragments, defects in the unfolded protein response, and decreased lifespan. RtcB also ligates endogenous pre-tRNA halves, and RtcB mutants have defects in growth and lifespan that can be bypassed by expression of pre-spliced tRNAs. In addition, animals that lack RtcB have defects that are independent of tRNA maturation and the unfolded protein response. Thus, RNA ligation by RtcB is required for the function of multiple endogenous target RNAs including both xbp-1 and tRNAs. RtcB is uniquely capable of performing these ligation functions, and RNA ligation by RtcB mediates multiple essential processes in vivo. Subject Categories Protein Biosynthesis & Quality Control; RNA Biology PMID:25366321

RPA-mediated unfolding of systematically varying G-quadruplex structures.

PubMed

Ray, Sujay; Qureshi, Mohammad H; Malcolm, Dominic W; Budhathoki, Jagat B; Celik, Uğur; Balci, Hamza

2013-05-21

G-quadruplex (GQ) is a noncanonical nucleic acid structure that is formed by guanine rich sequences. Unless it is destabilized by proteins such as replication protein A (RPA), GQ could interfere with DNA metabolic functions, such as replication or repair. We studied RPA-mediated GQ unfolding using single-molecule FRET on two groups of GQ structures that have different loop lengths and different numbers of G-tetrad layers. We observed a linear increase in the steady-state stability of the GQ against RPA-mediated unfolding with increasing number of layers or decreasing loop length. The stability demonstrated by different GQ structures varied by at least three orders of magnitude. Those with shorter loops (less than three nucleotides long) or a greater number of layers (more than three layers) maintained a significant folded population even at physiological RPA concentration (≈1 μM), raising the possibility of physiological viability of such GQ structures. Finally, we measured the transition time between the start and end of the RPA-mediated GQ unfolding process to be 0.35 ± 0.10 s for all GQ constructs we studied, despite significant differences in their steady-state stabilities. We propose a two-step RPA-mediated GQ unfolding mechanism that is consistent with our observations. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

RPA-Mediated Unfolding of Systematically Varying G-Quadruplex Structures

PubMed Central

Ray, Sujay; Qureshi, Mohammad H.; Malcolm, Dominic W.; Budhathoki, Jagat B.; Çelik, Uğur; Balci, Hamza

2013-01-01

G-quadruplex (GQ) is a noncanonical nucleic acid structure that is formed by guanine rich sequences. Unless it is destabilized by proteins such as replication protein A (RPA), GQ could interfere with DNA metabolic functions, such as replication or repair. We studied RPA-mediated GQ unfolding using single-molecule FRET on two groups of GQ structures that have different loop lengths and different numbers of G-tetrad layers. We observed a linear increase in the steady-state stability of the GQ against RPA-mediated unfolding with increasing number of layers or decreasing loop length. The stability demonstrated by different GQ structures varied by at least three orders of magnitude. Those with shorter loops (less than three nucleotides long) or a greater number of layers (more than three layers) maintained a significant folded population even at physiological RPA concentration (≈1 μM), raising the possibility of physiological viability of such GQ structures. Finally, we measured the transition time between the start and end of the RPA-mediated GQ unfolding process to be 0.35 ± 0.10 s for all GQ constructs we studied, despite significant differences in their steady-state stabilities. We propose a two-step RPA-mediated GQ unfolding mechanism that is consistent with our observations. PMID:23708363

Binding polarity of RPA to telomeric sequences and influence of G-quadruplex stability.

PubMed

Safa, Layal; Delagoutte, Emmanuelle; Petruseva, Irina; Alberti, Patrizia; Lavrik, Olga; Riou, Jean-François; Saintomé, Carole

2014-08-01

Replication protein A (RPA) is a single-stranded DNA binding protein that plays an essential role in telomere maintenance. RPA binds to and unfolds G-quadruplex (G4) structures formed in telomeric DNA, thus facilitating lagging strand DNA replication and telomerase activity. To investigate the effect of G4 stability on the interactions with human RPA (hRPA), we used a combination of biochemical and biophysical approaches. Our data revealed an inverse relationship between G4 stability and ability of hRPA to bind to telomeric DNA; notably small G4 ligands that enhance G4 stability strongly impaired G4 unfolding by hRPA. To gain more insight into the mechanism of binding and unfolding of telomeric G4 structures by RPA, we carried out photo-crosslinking experiments to elucidate the spatial arrangement of the RPA subunits along the DNA strands. Our results showed that RPA1 and RPA2 are arranged from 5' to 3' along the unfolded telomeric G4, as already described for unstructured single-stranded DNA, while no contact is possible with RPA3 on this short oligonucleotide. In addition, these data are compatible with a 5' to 3' directionality in G4 unfolding by hRPA. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Leaf fossils of the ancient Tasmanian relict Microcachrys (Podocarpaceae) from New Zealand.

PubMed

Carpenter, Raymond J; Jordan, Gregory J; Mildenhall, Dallas C; Lee, Daphne E

2011-07-01

Microcachrys tetragona (Podocarpaceae), endemic to the mountains of Tasmania, represents the only remaining taxon of one of the world's most ancient and widely distributed conifer lineages. Remarkably, however, despite its ∼150 Myr heritage, our understanding of the fossil history of this lineage is based almost entirely on the pollen record. Fossils of Microcachrys are especially important in light of recent molecular phylogenetic and dating evidence. This evidence dates the Microcachrys lineage to the Mesozoic and does not support the traditional placement of Microcachrys as sister to the southeastern Australian genus Pherosphaera. We undertook comparative studies of the foliage architecture, cuticle, and paleoecology of newly discovered fossils from the Oligo-Miocene of New Zealand and M. tetragona and discussed the importance of Microcachrys in the context of Podocarpaceae phylogeny. The fossils represent the oldest and first extra-Australian macrofossils of Microcachrys and are described as the new foliage species M. novae-zelandiae. These fossils confirm that the distinctive opposite decussate phyllotaxy of the genus is at least as old as the Oligo-Miocene and contribute to evidence that Microcachrys plants were sometimes important components of oligotrophic swampy habitats. Leaf fossils of Microcachrys closely comparable with the only extant species confirm that this lineage had a much wider past distribution. The fossil record and recent molecular phylogenetic studies, including that of Microcachrys, also serve to emphasize the important status of Tasmania as a refugium for seed plant lineages.

Spatial and temporal changes in leaf coloring date of Acer palmatum and Ginkgo biloba in response to temperature increases in South Korea.

PubMed

Park, Chang-Kyun; Ho, Chang-Hoi; Jeong, Su-Jong; Lee, Eun Ju; Kim, Jinwon

2017-01-01

Understanding shifts in autumn phenology associated with climate changes is critical for preserving forest ecosystems. This study examines the changes in the leaf coloring date (LCD) of two temperate deciduous tree species, Acer palmatum (Acer) and Ginkgo biloba (Ginkgo), in response to surface air temperature (Ts) changes at 54 stations of South Korea for the period 1989-2007. The variations of Acer and Ginkgo in South Korea are very similar: they show the same mean LCD of 295th day of the year and delays of about 0.45 days year-1 during the observation period. The delaying trend is closely correlated (correlation coefficient > 0.77) with increases in Ts in mid-autumn by 2.8 days °C-1. It is noted that the LCD delaying and temperature sensitivity (days °C-1) for both tree species show negligible dependences on latitudes and elevations. Given the significant LCD-Ts relation, we project LCD changes for 2016-35 and 2046-65 using a process-based model forced by temperature from climate model simulation. The projections indicate that the mean LCD would be further delayed by 3.2 (3.7) days in 2016-35 (2046-65) due to mid-autumn Ts increases. This study suggests that the mid-autumn warming is largely responsible for the observed LCD changes in South Korea and will intensify the delaying trends in the future.

Hypolipidemic and cardioprotective benefits of a novel fireberry hawthorn fruit extract in the JCR:LA-cp rodent model of dyslipidemia and cardiac dysfunction.

PubMed

Diane, Abdoulaye; Borthwick, Faye; Wu, Sheng; Lee, Jeanette; Brown, Paula N; Dickinson, Timothy A; Croft, Kevin D; Vine, Donna F; Proctor, Spencer D

2016-09-14

Hawthorn is a widely used herbal alternative medicine for the treatment of various cardiovascular diseases. However, the attributed health benefits, purported to be due to the presence of phenolic compounds, may depend on both the specific species and plant part. Studies to date investigating effects of hawthorn on heart disease(s) have used well-described European and/or Asian species, while little is known regarding the bioactivity of species native to North America. Six weeks of supplementation of both fireberry hawthorn berry (native Crataegus chrysocarpa) and English hawthorn leaf (C. monogyna, naturalized in North America) in the JCR:LA-cp rat, resulted in a significant reduction in heart weight, fasting LDL-C and improved heart function (p < 0.05). Fasting triglyceride and myocardial fibrosis were also reduced, but only by the berry extract. We demonstrate that both of the Canadian-sourced hawthorn extracts (introduced leaf and native berry) have cardioprotective benefits, likely via increased availability of nitric oxide.

Role of cavities and hydration in the pressure unfolding of T4 lysozyme

PubMed Central

Nucci, Nathaniel V.; Fuglestad, Brian; Athanasoula, Evangelia A.; Wand, A. Joshua

2014-01-01

It is well known that high hydrostatic pressures can induce the unfolding of proteins. The physical underpinnings of this phenomenon have been investigated extensively but remain controversial. Changes in solvation energetics have been commonly proposed as a driving force for pressure-induced unfolding. Recently, the elimination of void volumes in the native folded state has been argued to be the principal determinant. Here we use the cavity-containing L99A mutant of T4 lysozyme to examine the pressure-induced destabilization of this multidomain protein by using solution NMR spectroscopy. The cavity-containing C-terminal domain completely unfolds at moderate pressures, whereas the N-terminal domain remains largely structured to pressures as high as 2.5 kbar. The sensitivity to pressure is suppressed by the binding of benzene to the hydrophobic cavity. These results contrast to the pseudo-WT protein, which has a residual cavity volume very similar to that of the L99A–benzene complex but shows extensive subglobal reorganizations with pressure. Encapsulation of the L99A mutant in the aqueous nanoscale core of a reverse micelle is used to examine the hydration of the hydrophobic cavity. The confined space effect of encapsulation suppresses the pressure-induced unfolding transition and allows observation of the filling of the cavity with water at elevated pressures. This indicates that hydration of the hydrophobic cavity is more energetically unfavorable than global unfolding. Overall, these observations point to a range of cooperativity and energetics within the T4 lysozyme molecule and illuminate the fact that small changes in physical parameters can significantly alter the pressure sensitivity of proteins. PMID:25201963

Precursory signatures of protein folding/unfolding: From time series correlation analysis to atomistic mechanisms

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

Hsu, P. J.; Lai, S. K., E-mail: sklai@coll.phy.ncu.edu.tw; Molecular Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan

Folded conformations of proteins in thermodynamically stable states have long lifetimes. Before it folds into a stable conformation, or after unfolding from a stable conformation, the protein will generally stray from one random conformation to another leading thus to rapid fluctuations. Brief structural changes therefore occur before folding and unfolding events. These short-lived movements are easily overlooked in studies of folding/unfolding for they represent momentary excursions of the protein to explore conformations in the neighborhood of the stable conformation. The present study looks for precursory signatures of protein folding/unfolding within these rapid fluctuations through a combination of three techniques: (1)more » ultrafast shape recognition, (2) time series segmentation, and (3) time series correlation analysis. The first procedure measures the differences between statistical distance distributions of atoms in different conformations by calculating shape similarity indices from molecular dynamics simulation trajectories. The second procedure is used to discover the times at which the protein makes transitions from one conformation to another. Finally, we employ the third technique to exploit spatial fingerprints of the stable conformations; this procedure is to map out the sequences of changes preceding the actual folding and unfolding events, since strongly correlated atoms in different conformations are different due to bond and steric constraints. The aforementioned high-frequency fluctuations are therefore characterized by distinct correlational and structural changes that are associated with rate-limiting precursors that translate into brief segments. Guided by these technical procedures, we choose a model system, a fragment of the protein transthyretin, for identifying in this system not only the precursory signatures of transitions associated with α helix and β hairpin, but also the important role played by weaker correlations in such protein folding dynamics.« less

Towards data warehousing and mining of protein unfolding simulation data.

PubMed

Berrar, Daniel; Stahl, Frederic; Silva, Candida; Rodrigues, J Rui; Brito, Rui M M; Dubitzky, Werner

2005-10-01

The prediction of protein structure and the precise understanding of protein folding and unfolding processes remains one of the greatest challenges in structural biology and bioinformatics. Computer simulations based on molecular dynamics (MD) are at the forefront of the effort to gain a deeper understanding of these complex processes. Currently, these MD simulations are usually on the order of tens of nanoseconds, generate a large amount of conformational data and are computationally expensive. More and more groups run such simulations and generate a myriad of data, which raises new challenges in managing and analyzing these data. Because the vast range of proteins researchers want to study and simulate, the computational effort needed to generate data, the large data volumes involved, and the different types of analyses scientists need to perform, it is desirable to provide a public repository allowing researchers to pool and share protein unfolding data. To adequately organize, manage, and analyze the data generated by unfolding simulation studies, we designed a data warehouse system that is embedded in a grid environment to facilitate the seamless sharing of available computer resources and thus enable many groups to share complex molecular dynamics simulations on a more regular basis. To gain insight into the conformational fluctuations and stability of the monomeric forms of the amyloidogenic protein transthyretin (TTR), molecular dynamics unfolding simulations of the monomer of human TTR have been conducted. Trajectory data and meta-data of the wild-type (WT) protein and the highly amyloidogenic variant L55P-TTR represent the test case for the data warehouse. Web and grid services, especially pre-defined data mining services that can run on or 'near' the data repository of the data warehouse, are likely to play a pivotal role in the analysis of molecular dynamics unfolding data.

Reshaping the folding energy landscape by chloride salt: impact on molten-globule formation and aggregation behavior of carbonic anhydrase.

PubMed

Borén, Kristina; Grankvist, Hannah; Hammarström, Per; Carlsson, Uno

2004-05-21

During chemical denaturation different intermediate states are populated or suppressed due to the nature of the denaturant used. Chemical denaturation by guanidine-HCl (GuHCl) of human carbonic anhydrase II (HCA II) leads to a three-state unfolding process (Cm,NI=1.0 and Cm,IU=1.9 M GuHCl) with formation of an equilibrium molten-globule intermediate that is stable at moderate concentrations of the denaturant (1-2 M) with a maximum at 1.5 M GuHCl. On the contrary, urea denaturation gives rise to an apparent two-state unfolding transition (Cm=4.4 M urea). However, 8-anilino-1-naphthalene sulfonate (ANS) binding and decreased refolding capacity revealed the presence of the molten globule in the middle of the unfolding transition zone, although to a lesser extent than in GuHCl. Cross-linking studies showed the formation of moderate oligomer sized (300 kDa) and large soluble aggregates (>1000 kDa). Inclusion of 1.5 M NaCl to the urea denaturant to mimic the ionic character of GuHCl leads to a three-state unfolding behavior (Cm,NI=3.0 and Cm,IU=6.4 M urea) with a significantly stabilized molten-globule intermediate by the chloride salt. Comparisons between NaCl and LiCl of the impact on the stability of the various states of HCA II in urea showed that the effects followed what could be expected from the Hofmeister series, where Li+ is a chaotropic ion leading to decreased stability of the native state. Salt addition to the completely urea unfolded HCA II also led to an aggregation prone unfolded state, that has not been observed before for carbonic anhydrase. Refolding from this state only provided low recoveries of native enzyme.

Mechanism of Unfolding of Human Prion Protein.

PubMed

Singh, Reman K; Chamachi, Neharika G; Chakrabarty, Suman; Mukherjee, Arnab

2017-01-26

Misfolding and aggregation of prion proteins are associated with several neurodegenerative diseases. Therefore, understanding the mechanism of the misfolding process is of enormous interest in the scientific community. It has been speculated and widely discussed that the native cellular prion protein (PrP C ) form needs to undergo substantial unfolding to a more stable PrP C* state, which may further oligomerize into the toxic scrapie (PrP Sc ) form. Here, we have studied the mechanism of the unfolding of the human prion protein (huPrP) using a set of extensive well-tempered metadynamics simulations. Through multiple microsecond-long metadynamics simulations, we find several possible unfolding pathways. We show that each pathway leads to an unfolded state of lower free energy than the native state. Thus, our study may point to the signature of a PrP C* form that corresponds to a global minimum on the conformational free-energy landscape. Moreover, we find that these global minima states do not involve an increased β-sheet content, as was assumed to be a signature of PrP Sc formation in previous simulation studies. We have further analyzed the origin of metastability of the PrP C form through free-energy surfaces of the chopped helical segments to show that the helices, particularly H2 and H3 of the prion protein, have the tendency to form either a random coil or a β-structure. Therefore, the secondary structural elements of the prion protein are only weakly stabilized by tertiary contacts and solvation forces so that relatively weak perturbations induced by temperature, pressure, pH, and so forth can lead to substantial unfolding with characteristics of intrinsically disordered proteins.

Mechanics of collective unfolding

NASA Astrophysics Data System (ADS)

Caruel, M.; Allain, J.-M.; Truskinovsky, L.

2015-03-01

Mechanically induced unfolding of passive crosslinkers is a fundamental biological phenomenon encountered across the scales from individual macro-molecules to cytoskeletal actin networks. In this paper we study a conceptual model of athermal load-induced unfolding and use a minimalistic setting allowing one to emphasize the role of long-range interactions while maintaining full analytical transparency. Our model can be viewed as a description of a parallel bundle of N bistable units confined between two shared rigid backbones that are loaded through a series spring. We show that the ground states in this model correspond to synchronized, single phase configurations where all individual units are either folded or unfolded. We then study the fine structure of the wiggly energy landscape along the reaction coordinate linking the two coherent states and describing the optimal mechanism of cooperative unfolding. Quite remarkably, our study shows the fundamental difference in the size and the structure of the folding-unfolding energy barriers in the hard (fixed displacements) and soft (fixed forces) loading devices which persists in the continuum limit. We argue that both, the synchronization and the non-equivalence of the mechanical responses in hard and soft devices, have their origin in the dominance of long-range interactions. We then apply our minimal model to skeletal muscles where the power-stroke in acto-myosin crossbridges can be interpreted as passive folding. A quantitative analysis of the muscle model shows that the relative rigidity of myosin backbone provides the long-range interaction mechanism allowing the system to effectively synchronize the power-stroke in individual crossbridges even in the presence of thermal fluctuations. In view of the prototypical nature of the proposed model, our general conclusions pertain to a variety of other biological systems where elastic interactions are mediated by effective backbones.

Precursory signatures of protein folding/unfolding: From time series correlation analysis to atomistic mechanisms

NASA Astrophysics Data System (ADS)

Hsu, P. J.; Cheong, S. A.; Lai, S. K.

2014-05-01

Folded conformations of proteins in thermodynamically stable states have long lifetimes. Before it folds into a stable conformation, or after unfolding from a stable conformation, the protein will generally stray from one random conformation to another leading thus to rapid fluctuations. Brief structural changes therefore occur before folding and unfolding events. These short-lived movements are easily overlooked in studies of folding/unfolding for they represent momentary excursions of the protein to explore conformations in the neighborhood of the stable conformation. The present study looks for precursory signatures of protein folding/unfolding within these rapid fluctuations through a combination of three techniques: (1) ultrafast shape recognition, (2) time series segmentation, and (3) time series correlation analysis. The first procedure measures the differences between statistical distance distributions of atoms in different conformations by calculating shape similarity indices from molecular dynamics simulation trajectories. The second procedure is used to discover the times at which the protein makes transitions from one conformation to another. Finally, we employ the third technique to exploit spatial fingerprints of the stable conformations; this procedure is to map out the sequences of changes preceding the actual folding and unfolding events, since strongly correlated atoms in different conformations are different due to bond and steric constraints. The aforementioned high-frequency fluctuations are therefore characterized by distinct correlational and structural changes that are associated with rate-limiting precursors that translate into brief segments. Guided by these technical procedures, we choose a model system, a fragment of the protein transthyretin, for identifying in this system not only the precursory signatures of transitions associated with α helix and β hairpin, but also the important role played by weaker correlations in such protein folding dynamics.

Unfolding of a model protein on ion exchange and mixed mode chromatography surfaces.

PubMed

Gospodarek, Adrian M; Hiser, Diana E; O'Connell, John P; Fernandez, Erik J

2014-08-15

Recent studies with proteins indicate that conformational changes and aggregation can occur during ion exchange chromatography (IEC). Such behavior is not usually expected, but could lead to decreased yield and product degradation from both IEC and multi mode chromatography (MMC) that has ligands of both hydrophobic and charged functionalities. In this study, we used hydrogen exchange mass spectrometry to investigate unfolding of the model protein BSA on IEC and MMC surfaces under different solution conditions at 25°C. Increased solvent exposure, indicating greater unfolding relative to that in solution, was found for protein adsorbed on cationic IEC and MMC surfaces in the pH range of 3.0 to 4.5, where BSA has decreased stability in solution. There was no effect of anionic surfaces at pH values in the range from 6.0 to 9.0. Differences of solvent exposure of whole molecules when adsorbed and in solution suggest that adsorbed BSA unfolds at lower pH values and may show aggregation, depending upon pH and the surface type. Measurements on digested peptides showed that classifications of stability can be made for various regions; these are generally retained as pH is changed. When salt was added to MMC systems, where electrostatic interactions would be minimized, less solvent exposure was seen, implying that it is the cationic moieties, rather than the hydrophobic ligands, which cause greater surface unfolding at low salt concentrations. These results suggest that proteins of lower stability may exhibit unfolding and aggregation during IEC and MMC separations, as they can with hydrophobic interaction chromatography. Copyright © 2014 Elsevier B.V. All rights reserved.

Dynamic properties of bacterial pili measured by optical tweezers

NASA Astrophysics Data System (ADS)

Fallman, Erik G.; Andersson, Magnus J.; Schedin, Staffan S.; Jass, Jana; Uhlin, Bernt Eric; Axner, Ove

2004-10-01

The ability of uropathogenic Escherichia coli (UPEC) to cause urinary tract infections is dependent on their ability to colonize the uroepithelium. Infecting bacteria ascend the urethra to the bladder and then kidneys by attaching to the uroepithelial cells via the differential expression of adhesins. P pili are associated with pyelonephritis, the more severe infection of the kidneys. In order to find means to treat pyelonephritis, it is therefore of interest to investigate the properties P pili. The mechanical behavior of individual P pili of uropathogenic Escherichia coli has recently been investigated using optical tweezers. P pili, whose main part constitutes the PapA rod, composed of ~1000 PapA subunits in a helical arrangement, are distributed over the bacterial surface and mediate adhesion to host cells. We have earlier studied P pili regarding its stretching/elongation properties where we have found and characterized three different elongation regions, of which one constitute an unfolding of the quaternary (helical) structure of the PapA rod. It was shown that this unfolding takes place at an elongation independent force of 27 +/- 2 pN. We have also recently performed studies on its folding properties and shown that the unfolding/folding of the PapA rod is completely reversible. Here we present a study of the dynamical properties of the PapA rod. We show, among other things, that the unfolding force increases and that the folding force decreases with the speed of unfolding and folding respectively. Moreover, the PapA rod can be folded-unfolded a significant number of times without loosing its characteristics, a phenomenon that is believed to be important for the bacterium to keep close contact to the host tissue and consequently helps the bacterium to colonize the host tissue.

Determining the folding and unfolding rate constants of nucleic acids by biosensor. Application to telomere G-quadruplex.

PubMed

Zhao, Yong; Kan, Zhong-yuan; Zeng, Zhi-xiong; Hao, Yu-hua; Chen, Hua; Tan, Zheng

2004-10-20

Nucleic acid molecules may fold into secondary structures, and the formation of such structures is involved in many biological processes and technical applications. The folding and unfolding rate constants define the kinetics of conformation interconversion and the stability of these structures and is important in realizing their functions. We developed a method to determine these kinetic parameters using an optical biosensor based on surface plasmon resonance. The folding and unfolding of a nucleic acid is coupled with a hybridization reaction by immobilization of the target nucleic acid on a sensor chip surface and injection of a complementary probe nucleic acid over the sensor chip surface. By monitoring the time course of duplex formation, both the folding and unfolding rate constants for the target nucleic acid and the association and dissociation rate constants for the target-probe duplex can all be derived from the same measurement. We applied this method to determine the folding and unfolding rate constants of the G-quadruplex of human telomere sequence (TTAGGG)(4) and its association and dissociation rate constants with the complementary strand (CCCTAA)(4). The results show that both the folding and unfolding occur on the time scale of minutes at physiological concentration of K(+). We speculate that this property might be important for telomere elongation. A complete set of the kinetic parameters for both of the structures allows us to study the competition between the formation of the quadruplex and the duplex. Calculations indicate that the formation of both the quadruplex and the duplex is strand concentration-dependent, and the quadruplex can be efficiently formed at low strand concentration. This property may provide the basis for the formation of the quadruplex in vivo in the presence of a complementary strand.

Denaturation of RNA secondary and tertiary structure by urea: simple unfolded state models and free energy parameters account for measured m-values

PubMed Central

Lambert, Dominic; Draper, David E.

2012-01-01

To investigate the mechanism by which urea destabilizes RNA structure, urea-induced unfolding of four different RNA secondary and tertiary structures was quantified in terms of an m-value, the rate at which the free energy of unfolding changes with urea molality. From literature data and our osmometric study of a backbone analog, we derived average interaction potentials (per Å2 of solvent accessible surface) between urea and three kinds of RNA surfaces: phosphate, ribose, and base. Estimates of the increases in solvent accessible surface areas upon RNA denaturation were based on a simple model of unfolded RNA as a combination of helical and single strand segments. These estimates, combined with the three interaction potentials and a term to account for urea interactions with released ions, yield calculated m-values in good agreement with experimental values (200 mm monovalent salt). Agreement was obtained only if single-stranded RNAs were modeled in a highly stacked, A form conformation. The primary driving force for urea induced denaturation is the strong interaction of urea with the large surface areas of bases that become exposed upon denaturation of either RNA secondary or tertiary structure, though urea interactions with backbone and released ions may account for up to a third of the m-value. Urea m-values for all four RNA are salt-dependent, which we attribute to an increased extension (or decreased charge density) of unfolded RNAs with increased urea concentration. The sensitivity of the urea m-value to base surface exposure makes it a potentially useful probe of the conformations of RNA unfolded states. PMID:23088364

Elucidating quantitative stability/flexibility relationships within thioredoxin and its fragments using a distance constraint model.

PubMed

Jacobs, Donald J; Livesay, Dennis R; Hules, Jeremy; Tasayco, Maria Luisa

2006-05-05

Numerous quantitative stability/flexibility relationships, within Escherichia coli thioredoxin (Trx) and its fragments are determined using a minimal distance constraint model (DCM). A one-dimensional free energy landscape as a function of global flexibility reveals Trx to fold in a low-barrier two-state process, with a voluminous transition state. Near the folding transition temperature, the native free energy basin is markedly skewed to allow partial unfolded forms. Under native conditions the skewed shape is lost, and the protein forms a compact structure with some flexibility. Predictions on ten Trx fragments are generally consistent with experimental observations that they are disordered, and that complementary fragments reconstitute. A hierarchical unfolding pathway is uncovered using an exhaustive computational procedure of breaking interfacial cross-linking hydrogen bonds that span over a series of fragment dissociations. The unfolding pathway leads to a stable core structure (residues 22-90), predicted to act as a kinetic trap. Direct connection between degree of rigidity within molecular structure and non-additivity of free energy is demonstrated using a thermodynamic cycle involving fragments and their hierarchical unfolding pathway. Additionally, the model provides insight about molecular cooperativity within Trx in its native state, and about intermediate states populating the folding/unfolding pathways. Native state cooperativity correlation plots highlight several flexibly correlated regions, giving insight into the catalytic mechanism that facilitates access to the active site disulfide bond. Residual native cooperativity correlations are present in the core substructure, suggesting that Trx can function when it is partly unfolded. This natively disordered kinetic trap, interpreted as a molten globule, has a wide temperature range of metastability, and it is identified as the "slow intermediate state" observed in kinetic experiments. These computational results are found to be in overall agreement with a large array of experimental data.

Correlating folding and signaling in a photoreceptor by single molecule measurements and energy landscape calculations

NASA Astrophysics Data System (ADS)

Hoff, Wouter

2007-03-01

Receptor activation is a fundamental process in biological signaling. We study the structural changes during activation of photoactive yellow protein (PYP). This is triggered by photoisomerization of the p-coumaric acid (pCA) chromophore of PYP, which converts the initial pG state into the activated pB state. Mechanical unfolding of Cys-linked PYP multimers probed by atomic force microscopy (AFM) in the presence and absence of illumination reveals that the core of the protein is extended by 3 nm and destabilized by 30 percent in pB. These results establish a generally applicable single molecule approach for mapping functional conformational changes to selected regions of a protein and indicate that stimulus-induced partial protein unfolding can be employed as a signaling mechanism. Comparative measurements, Jarzynski-Hummer-Szabo analysis of the data, and steered MD simulations of two double-Cys PYP mutants reveal strong anisotropy in the unfolding mechanism along the two axes defined by the Cys residues. Unfolding along one axis exhibits a transition-state-like feature where six hydrogen bonds break simultaneously. The other axis displays an unpeaked force profile reflecting a non-cooperative transition, challenging the notion that cooperative unfolding is a universal feature in protein stability. MD simulations with a coarse-grained protein model show that the folding of pG is two-state, consistent with experimental observations. In contrast, the folding free energy surface of a coarse-grained model of pB involves an on-pathway partially unfolded intermediate that closely matches experimental data. The results reveal that interactions between the pCA and its binding pocket can switch the energy landscape for PYP from two- to three-state folding, and show how this can be exploited to trigger large functionally important protein conformational changes.

Protein structural dynamics at the gas/water interface examined by hydrogen exchange mass spectrometry.

PubMed

Xiao, Yiming; Konermann, Lars

2015-08-01

Gas/water interfaces (such as air bubbles or foam) are detrimental to the stability of proteins, often causing aggregation. This represents a potential problem for industrial processes, for example, the production and handling of protein drugs. Proteins possess surfactant-like properties, resulting in a high affinity for gas/water interfaces. The tendency of previously buried nonpolar residues to maximize contact with the gas phase can cause significant structural distortion. Most earlier studies in this area employed spectroscopic tools that could only provide limited information. Here we use hydrogen/deuterium exchange (HDX) mass spectrometry (MS) for probing the conformational dynamics of the model protein myoglobin (Mb) in the presence of N(2) bubbles. HDX/MS relies on the principle that unfolded and/or highly dynamic regions undergo faster deuteration than tightly folded segments. In bubble-free solution Mb displays EX2 behavior, reflecting the occurrence of short-lived excursions to partially unfolded conformers. A dramatically different behavior is seen in the presence of N(2) bubbles; EX2 dynamics still take place, but in addition the protein shows EX1 behavior. The latter results from interconversion of the native state with conformers that are globally unfolded and long-lived. These unfolded species likely correspond to Mb that is adsorbed to the surface of gas bubbles. N(2) sparging also induces aggregation. To explain the observed behavior we propose a simple model, that is, "semi-unfolded" ↔ "native" ↔ "globally unfolded" → "aggregated". This model quantitatively reproduces the experimentally observed kinetics. To the best of our knowledge, the current study marks the first exploration of surface denaturation phenomena by HDX/MS. © 2015 The Protein Society.

A revised checklist of Nepticulidae fossils (Lepidoptera) indicates an Early Cretaceous origin.

PubMed

Doorenweerd, Camiel; Nieukerken, Erik J Van; Sohn, Jae-Cheon; Labandeira, Conrad C

2015-05-27

With phylogenetic knowledge of Lepidoptera rapidly increasing, catalysed by increasingly powerful molecular techniques, the demand for fossil calibration points to estimate an evolutionary timeframe for the order is becoming an increasingly pressing issue. The family Nepticulidae is a species rich, basal branch within the phylogeny of the Lepidoptera, characterized by larval leaf-mining habits, and thereby represents a potentially important lineage whose evolutionary history can be established more thoroughly with the potential use of fossil calibration points. Using our experience with extant global Nepticulidae, we discuss a list of characters that may be used to assign fossil leaf mines to Nepticulidae, and suggest useful methods for classifying relevant fossil material. We present a checklist of 79 records of Nepticulidae representing adult and leaf-mine fossils mentioned in literature, often with multiple exemplars constituting a single record. We provide our interpretation of these fossils. Two species now are included in the collective generic name Stigmellites: Stigmellites resupinata (Krassilov, 2008) comb. nov. (from Ophiheliconoma) and Stigmellites almeidae (Martins-Neto, 1989) comb. nov. (from Nepticula). Eleven records are for the first time attributed to Nepticulidae. After discarding several dubious records, including one possibly placing the family at a latest Jurassic position, we conclude that the oldest fossils likely attributable to Nepticulidae are several exemplars representing a variety of species from the Dakota Formation (USA). The relevant strata containing these earliest fossils are now dated at 102 Ma (million years ago) in age, corresponding to the latest Albian Stage of the Early Cretaceous. Integration of all records in the checklist shows that a continuous presence of nepticulid-like leaf mines preserved as compression-impression fossils and by amber entombment of adults have a fossil record extending to the latest Early Cretaceous.

Mapping the energy landscape for second-stage folding of a single membrane protein

PubMed Central

Min, Duyoung; Jefferson, Robert E; Bowie, James U; Yoon, Tae-Young

2016-01-01

Membrane proteins are designed to fold and function in a lipid membrane, yet folding experiments within a native membrane environment are challenging to design. Here we show that single-molecule forced unfolding experiments can be adapted to study helical membrane protein folding under native-like bicelle conditions. Applying force using magnetic tweezers, we find that a transmembrane helix protein, Escherichia coli rhomboid protease GlpG, unfolds in a highly cooperative manner, largely unraveling as one physical unit in response to mechanical tension above 25 pN. Considerable hysteresis is observed, with refolding occurring only at forces below 5 pN. Characterizing the energy landscape reveals only modest thermodynamic stability (ΔG = 6.5 kBT) but a large unfolding barrier (21.3 kBT) that can maintain the protein in a folded state for long periods of time (t1/2 ~3.5 h). The observed energy landscape may have evolved to limit the existence of troublesome partially unfolded states and impart rigidity to the structure. PMID:26479439

An unfolded protein-induced conformational switch activates mammalian IRE1

PubMed Central

Acosta-Alvear, Diego; Nguyen, Hieu T; Lee, Crystal P; Chu, Feixia

2017-01-01

The unfolded protein response (UPR) adjusts the cell’s protein folding capacity in the endoplasmic reticulum (ER) according to need. IRE1 is the most conserved UPR sensor in eukaryotic cells. It has remained controversial, however, whether mammalian and yeast IRE1 use a common mechanism for ER stress sensing. Here, we show that similar to yeast, human IRE1α’s ER-lumenal domain (hIRE1α LD) binds peptides with a characteristic amino acid bias. Peptides and unfolded proteins bind to hIRE1α LD’s MHC-like groove and induce allosteric changes that lead to its oligomerization. Mutation of a hydrophobic patch at the oligomerization interface decoupled peptide binding to hIRE1α LD from its oligomerization, yet retained peptide-induced allosteric coupling within the domain. Importantly, impairing oligomerization of hIRE1α LD abolished IRE1’s activity in living cells. Our results provide evidence for a unifying mechanism of IRE1 activation that relies on unfolded protein binding-induced oligomerization. PMID:28971800

Unfolding four-helix bundles

NASA Astrophysics Data System (ADS)

Gray, Harry B.; Winkler, Jay R.; Kozak, John J.

2011-03-01

A geometrical model has been developed to describe the early stages of unfolding of cytochromes c‧ and c-b562 . Calculations are based on a step-wise extension of the polypeptide chain subject to the constraint that the spatial relationship among the residues of each triplet is fixed by the native-state crystallographic data. The response of each protein to these structural perturbations allows the evolution of each of the four helices in these two proteins to be differentiated. It is found that the two external helices in c‧ unfold before its two internal helices, whereas exactly the opposite behaviour is demonstrated by c-b562 . Each of these cytochromes has an extended, internal, non-helical ('turning') region that initially lags behind the most labile helix but then, at a certain stage (identified for each cytochrome), unravels before any of the four helices present in the native structure. It is believed that these predictions will be useful in guiding future experimental studies on the unfolding of these two cytochromes.

Molecular dynamics simulations and CD spectroscopy reveal hydration-induced unfolding of the intrinsically disordered LEA proteins COR15A and COR15B from Arabidopsis thaliana.

PubMed

Navarro-Retamal, Carlos; Bremer, Anne; Alzate-Morales, Jans; Caballero, Julio; Hincha, Dirk K; González, Wendy; Thalhammer, Anja

2016-10-07

The LEA (late embryogenesis abundant) proteins COR15A and COR15B from Arabidopsis thaliana are intrinsically disordered under fully hydrated conditions, but obtain α-helical structure during dehydration, which is reversible upon rehydration. To understand this unusual structural transition, both proteins were investigated by circular dichroism (CD) and molecular dynamics (MD) approaches. MD simulations showed unfolding of the proteins in water, in agreement with CD data obtained with both HIS-tagged and untagged recombinant proteins. Mainly intramolecular hydrogen bonds (H-bonds) formed by the protein backbone were replaced by H-bonds with water molecules. As COR15 proteins function in vivo as protectants in leaves partially dehydrated by freezing, unfolding was further assessed under crowded conditions. Glycerol reduced (40%) or prevented (100%) unfolding during MD simulations, in agreement with CD spectroscopy results. H-bonding analysis indicated that preferential exclusion of glycerol from the protein backbone increased stability of the folded state.

Pharmacological chaperone reshapes the energy landscape for folding and aggregation of the prion protein

NASA Astrophysics Data System (ADS)

Gupta, Amar Nath; Neupane, Krishna; Rezajooei, Negar; Cortez, Leonardo M.; Sim, Valerie L.; Woodside, Michael T.

2016-06-01

The development of small-molecule pharmacological chaperones as therapeutics for protein misfolding diseases has proven challenging, partly because their mechanism of action remains unclear. Here we study Fe-TMPyP, a tetrapyrrole that binds to the prion protein PrP and inhibits misfolding, examining its effects on PrP folding at the single-molecule level with force spectroscopy. Single PrP molecules are unfolded with and without Fe-TMPyP present using optical tweezers. Ligand binding to the native structure increases the unfolding force significantly and alters the transition state for unfolding, making it more brittle and raising the barrier height. Fe-TMPyP also binds the unfolded state, delaying native refolding. Furthermore, Fe-TMPyP binding blocks the formation of a stable misfolded dimer by interfering with intermolecular interactions, acting in a similar manner to some molecular chaperones. The ligand thus promotes native folding by stabilizing the native state while also suppressing interactions driving aggregation.

Unfolding the neutron spectrum of a NE213 scintillator using artificial neural networks.

PubMed

Sharghi Ido, A; Bonyadi, M R; Etaati, G R; Shahriari, M

2009-10-01

Artificial neural networks technology has been applied to unfold the neutron spectra from the pulse height distribution measured with NE213 liquid scintillator. Here, both the single and multi-layer perceptron neural network models have been implemented to unfold the neutron spectrum from an Am-Be neutron source. The activation function and the connectivity of the neurons have been investigated and the results have been analyzed in terms of the network's performance. The simulation results show that the neural network that utilizes the Satlins transfer function has the best performance. In addition, omitting the bias connection of the neurons improve the performance of the network. Also, the SCINFUL code is used for generating the response functions in the training phase of the process. Finally, the results of the neural network simulation have been compared with those of the FORIST unfolding code for both (241)Am-Be and (252)Cf neutron sources. The results of neural network are in good agreement with FORIST code.

Web-based unfolding cases: a strategy to enhance and evaluate clinical reasoning skills.

PubMed

Johnson, Gail; Flagler, Susan

2013-10-01

Clinical reasoning involves the use of both analytical and nonanalytical intuitive cognitive processes. Fostering student development of clinical reasoning skills and evaluating student performance in this cognitive arena can challenge educators. The use of Web-based unfolding cases is proposed as a strategy to address these challenges. Unfolding cases mimic real-life clinical situations by presenting only partial clinical information in sequential segments. Students receive immediate feedback after submitting a response to a given segment. The student's comparison of the desired and submitted responses provides information to enhance the development of clinical reasoning skills. Each student's set of case responses are saved for the instructor in an individual-student electronic file, providing a record of the student's knowledge and thinking processes for faculty evaluation. For the example case given, the approaches used to evaluate individual components of clinical reasoning are provided. Possible future uses of Web-based unfolding cases are described. Copyright 2013, SLACK Incorporated.

Initiation of Phage Infection by Partial Unfolding and Prolyl Isomerization*♦

PubMed Central

Hoffmann-Thoms, Stephanie; Weininger, Ulrich; Eckert, Barbara; Jakob, Roman P.; Koch, Johanna R.; Balbach, Jochen; Schmid, Franz X.

2013-01-01

Infection of Escherichia coli by the filamentous phage fd starts with the binding of the N2 domain of the phage gene-3-protein to an F pilus. This interaction triggers partial unfolding of the gene-3-protein, cis → trans isomerization at Pro-213, and domain disassembly, thereby exposing its binding site for the ultimate receptor TolA. The trans-proline sets a molecular timer to maintain the binding-active state long enough for the phage to interact with TolA. We elucidated the changes in structure and local stability that lead to partial unfolding and thus to the activation of the gene-3-protein for phage infection. Protein folding and TolA binding experiments were combined with real-time NMR spectroscopy, amide hydrogen exchange measurements, and phage infectivity assays. In combination, the results provide a molecular picture of how a local unfolding reaction couples with prolyl isomerization not only to generate the activated state of a protein but also to maintain it for an extended time. PMID:23486474

The small angle x-ray scattering of globular proteins in solution during heat denaturation

NASA Astrophysics Data System (ADS)

Banuelos, Jose; Urquidi, Jacob

2008-10-01

The ability of proteins to change their conformation in response to changes in their environment has consequences in biological processes like metabolism, chemical regulation in cells, and is believed to play a role in the onset of several neurodegenerative diseases. Factors such as a change in temperature, pressure, and the introduction of ions into the aqueous environment of a protein can give rise to the folding/unfolding of a protein. As a protein unfolds, the ratio of nonpolar to polar groups exposed to water changes, affecting a protein's thermodynamic properties. Using small angle x-ray scattering (SAXS), we are currently studying the intermediate protein conformations that arise during the folding/unfolding process as a function of temperature for five globular proteins. Trends in the observed intermediate structures of these globular proteins, along with correlations with data on protein thermodynamics may help elucidate shared characteristics between all proteins in the folding/unfolding process. Experimental design considerations will be discussed and preliminary results for some of these systems will be presented.

Unfolding of a ClC chloride transporter retains memory of its evolutionary history.

PubMed

Min, Duyoung; Jefferson, Robert E; Qi, Yifei; Wang, Jing Yang; Arbing, Mark A; Im, Wonpil; Bowie, James U

2018-05-01

ClC chloride channels and transporters are important for chloride homeostasis in species from bacteria to human. Mutations in ClC proteins cause genetically inherited diseases, some of which are likely to involve folding defects. The ClC proteins present a challenging and unusual biological folding problem because they are large membrane proteins possessing a complex architecture, with many reentrant helices that go only partway through membrane and loop back out. Here we were able to examine the unfolding of the Escherichia coli ClC transporter, ClC-ec1, using single-molecule forced unfolding methods. We found that the protein could be separated into two stable halves that unfolded independently. The independence of the two domains is consistent with an evolutionary model in which the two halves arose from independently folding subunits that later fused together. Maintaining smaller folding domains of lesser complexity within large membrane proteins may be an advantageous strategy to avoid misfolding traps.

Contribution of long-range interactions to the secondary structure of an unfolded globin.

PubMed

Fedyukina, Daria V; Rajagopalan, Senapathy; Sekhar, Ashok; Fulmer, Eric C; Eun, Ye-Jin; Cavagnero, Silvia

2010-09-08

This work explores the effect of long-range tertiary contacts on the distribution of residual secondary structure in the unfolded state of an alpha-helical protein. N-terminal fragments of increasing length, in conjunction with multidimensional nuclear magnetic resonance, were employed. A protein representative of the ubiquitous globin fold was chosen as the model system. We found that, while most of the detectable alpha-helical population in the unfolded ensemble does not depend on the presence of the C-terminal region (corresponding to the native G and H helices), specific N-to-C long-range contacts between the H and A-B-C regions enhance the helical secondary structure content of the N terminus (A-B-C regions). The simple approach introduced here, based on the evaluation of N-terminal polypeptide fragments of increasing length, is of general applicability to identify the influence of long-range interactions in unfolded proteins. Copyright 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.

ATP-induced noncooperative thermal unfolding of hen lysozyme

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

Liu, Honglin; Yin, Peidong; He, Shengnan

To understand the role of ATP underlying the enhanced amyloidosis of hen egg white lysozyme (HEWL), the synchrotron radiation circular dichroism, combined with tryptophan fluorescence, dynamic light-scattering, and differential scanning calorimetry, is used to examine the alterations of the conformation and thermal unfolding pathway of the HEWL in the presence of ATP, Mg{sup 2+}-ATP, ADP, AMP, etc. It is revealed that the binding of ATP to HEWL through strong electrostatic interaction changes the secondary structures of HEWL and makes the exposed residue W62 move into hydrophobic environments. This alteration of W62 decreases the {beta}-domain stability of HEWL, induces a noncooperativemore » unfolding of the secondary structures, and produces a partially unfolded intermediate. This intermediate containing relatively rich {alpha}-helix and less {beta}-sheet structures has a great tendency to aggregate. The results imply that the ease of aggregating of HEWL is related to the extent of denaturation of the amyloidogenic region, rather than the electrostatic neutralizing effect or monomeric {beta}-sheet enriched intermediate.« less

Estimation of neutron spectrum in the low-level gamma spectroscopy system using unfolding procedure

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

Knežević, D., E-mail: david.knezevic@df.uns.ac.rs; Jovančević, N.; Krmar, M.

2016-03-25

The radiation resulting from neutron interactions with Ge nuclei in active volume of HPGe detectors is one of the main concerns in low-level gamma spectroscopy measurements [1,2]. It is usually not possible to measure directly spectrum of neutrons which strike detector. This paper explore the possibility of estimation of neutron spectrum using measured activities of certain Ge(n,γ) and Ge(n,n’) reactions (obtained from low-level gamma measurements), available ENDF cross section data and unfolding procedures. In this work HPGe detector with passive shield made from commercial low background lead was used for the measurement. The most important objective of this study wasmore » to reconstruct muon induced neutron spectrum created in the shield of the HPGe detector. MAXED [3] and GRAVEL [4] algorithms for neutron spectra unfolding were used. The results of those two algorithms were compared and we analyzed the sensitivity of the unfolding procedure to the various input parameters.« less

Geospatial Method for Computing Supplemental Multi-Decadal U.S. Coastal Land-Use and Land-Cover Classification Products, Using Landsat Data and C-CAP Products

NASA Technical Reports Server (NTRS)

Spruce, J. P.; Smoot, James; Ellis, Jean; Hilbert, Kent; Swann, Roberta

2012-01-01

This paper discusses the development and implementation of a geospatial data processing method and multi-decadal Landsat time series for computing general coastal U.S. land-use and land-cover (LULC) classifications and change products consisting of seven classes (water, barren, upland herbaceous, non-woody wetland, woody upland, woody wetland, and urban). Use of this approach extends the observational period of the NOAA-generated Coastal Change and Analysis Program (C-CAP) products by almost two decades, assuming the availability of one cloud free Landsat scene from any season for each targeted year. The Mobile Bay region in Alabama was used as a study area to develop, demonstrate, and validate the method that was applied to derive LULC products for nine dates at approximate five year intervals across a 34-year time span, using single dates of data for each classification in which forests were either leaf-on, leaf-off, or mixed senescent conditions. Classifications were computed and refined using decision rules in conjunction with unsupervised classification of Landsat data and C-CAP value-added products. Each classification's overall accuracy was assessed by comparing stratified random locations to available reference data, including higher spatial resolution satellite and aerial imagery, field survey data, and raw Landsat RGBs. Overall classification accuracies ranged from 83 to 91% with overall Kappa statistics ranging from 0.78 to 0.89. The accuracies are comparable to those from similar, generalized LULC products derived from C-CAP data. The Landsat MSS-based LULC product accuracies are similar to those from Landsat TM or ETM+ data. Accurate classifications were computed for all nine dates, yielding effective results regardless of season. This classification method yielded products that were used to compute LULC change products via additive GIS overlay techniques.

Hyperspectral and Thermal Imaging of Oilseed Rape (Brassica napus) Response to Fungal Species of the Genus Alternaria

PubMed Central

Baranowski, Piotr; Jedryczka, Malgorzata; Mazurek, Wojciech; Babula-Skowronska, Danuta; Siedliska, Anna; Kaczmarek, Joanna

2015-01-01

In this paper, thermal (8-13 µm) and hyperspectral imaging in visible and near infrared (VNIR) and short wavelength infrared (SWIR) ranges were used to elaborate a method of early detection of biotic stresses caused by fungal species belonging to the genus Alternaria that were host (Alternaria alternata, Alternaria brassicae, and Alternaria brassicicola) and non-host (Alternaria dauci) pathogens to oilseed rape (Brassica napus L.). The measurements of disease severity for chosen dates after inoculation were compared to temperature distributions on infected leaves and to averaged reflectance characteristics. Statistical analysis revealed that leaf temperature distributions on particular days after inoculation and respective spectral characteristics, especially in the SWIR range (1000-2500 nm), significantly differed for the leaves inoculated with A. dauci from the other species of Alternaria as well as from leaves of non-treated plants. The significant differences in leaf temperature of the studied Alternaria species were observed in various stages of infection development. The classification experiments were performed on the hyperspectral data of the leaf surfaces to distinguish days after inoculation and Alternaria species. The second-derivative transformation of the spectral data together with back-propagation neural networks (BNNs) appeared to be the best combination for classification of days after inoculation (prediction accuracy 90.5%) and Alternaria species (prediction accuracy 80.5%). PMID:25826369