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Sample records for exploiting disulfide assisted

  1. Redox-Innocent Metal-Assisted Cleavage of S-S Bond in a Disulfide-Containing Ligand.

    PubMed

    Esmieu, Charlène; Orio, Maylis; Le Pape, Laurent; Lebrun, Colette; Pécaut, Jacques; Ménage, Stéphane; Torelli, Stéphane

    2016-06-20

    Due to their redox capabilities, thiols have an important role in biological oxidative/reductive processes through the formation of disulfides or their oxidation to into sulfenic, sulfinic, or sulfonic derivatives being also relevant for specific enzyme activities. The mechanisms of these biological pathways often involve metal ion(s). In this case, deciphering metal-assisted transformation of the S-S bond is of primary interest. This report details the reactivity of the disulfide-containing 2,6-bis[(bis(pyridylmethyl)amino)methyl]-4-methylmercaptophenyldisulfide (L(Me(BPA)S-S)) ligand with Cu(II) using different experimental conditions (anaerobic, H2O-only, H2O/O2, or O2-only). Crystallographic snapshots show the formation of tetranuclear disulfide, dinuclear sulfinate, and sulfonate complexes. Mechanistic investigations using Zn(II) as control indicate a non-metal-redox-assisted process in all cases. When present, water acts as nucleophile and attacks at the S-S bond. Under anhydrous conditions, a different pathway involving a direct O2 attack at the disulfide is proposed. PMID:27268152

  2. Surface modification of amorphous substrates by disulfide derivatives: A photo-assisted route to direct functionalization of chalcogenide glasses

    NASA Astrophysics Data System (ADS)

    Amalric, Julien; Marchand-Brynaert, Jacqueline

    2011-12-01

    A novel route for chalcogenide glass surface modification is disclosed. The formation of an organic monolayer from disulfide derivatives is studied on two different glasses of formula GexAsySez by water contact angle measurement, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy in attenuated total reflection mode (FTIR-ATR). The potential anchoring group is the disulfide functionality. Since thioctic acid derivatives absorb around 335 nm, an irradiation step is included, in order to favor S-S disruption. Three types of disulfide compounds are grafted onto small glass breaks for contact angle and XPS analyses. The results show effective changes of surface state. According to contact angle measurement, the deposited organic layer functionalized by a small polyethylene glycol chain leads to a more hydrophilic surface, long alkyl chain or a perfluorinated carbon chain leads to a more hydrophobic surface. XPS shows the presence at the surface of an organic layer with sulfur and ethylene oxide chains, or augmentation of organic carbons or fluorine and Csbnd F bonds. The photo-assisted grafting of the disulfides onto an ATR prism made of chalcogenide glass shows that this surface modification process does not affect infrared transparency, despite UV treatment, and accurate structural analysis can be performed.

  3. An Educational Program to Assist Clinicians in Identifying Elder Investment Fraud and Financial Exploitation

    ERIC Educational Resources Information Center

    Mills, Whitney L.; Roush, Robert E.; Moye, Jennifer; Kunik, Mark E.; Wilson, Nancy L.; Taffet, George E.; Naik, Aanand D.

    2012-01-01

    Due to age-related factors and illnesses, older adults may become vulnerable to elder investment fraud and financial exploitation (EIFFE). The authors describe the development and preliminary evaluation of an educational program to raise awareness and assist clinicians in identifying older adults at risk. Participants (n = 127) gave high ratings…

  4. A metal-ion-assisted assembly approach to synthesize disulfide-bridged periodical mesoporous organosilicas with high sulfide contents and efficient adsorption

    NASA Astrophysics Data System (ADS)

    Hao, Na; Han, Lu; Yang, Yunxia; Wang, Huanting; Webley, Paul A.; Zhao, Dongyuan

    2010-06-01

    Well-ordered two-dimensional (2D) hexagonal periodic mesoporous organosilicas (PMOs) with a high content of disulfide groups have been prepared by a simple metal-ions-assisted amphiphilic surfactant templating process under a strong acidic condition. Long-chain organic bridge silane, bis(triethoxysilylpropyl)disulfide (BTSPDS) was used as a precursor which can be co-condensed with tetraethoxysilane (TEOS) to assemble with the triblock copolymer Pluronic P123 template and to construct the mesostructured organic-inorganic frameworks. The content of disulfide functional groups is up to 20% (BTSPDS molar content in the initial silane mixture) incorporated into the framework. The obtained ordered mesoporous DS-PMO materials have relatively high BET surface area (˜580 m 2/g), large uniform pore size (up to 6.3 nm) and thick pore walls (thickness up to 7.1 nm), because of the long-chain disulfide bridges. The metal ions such as Zn 2+ formed the four-coordination complex with two sulfides of BTSPDS and ethylene oxide moieties of P123 template, which could enhance the interaction between the "soft" long disulfide groups and P123 template, thus improving the mesostructural regularity correspondingly. The disulfide-bridged PMO materials exhibit excellent hydrothermal stability in boiling water for 5 days, probably due to the thick pore walls. SEM images show that after the hydrothermal treatment, the morphology of the ordered disulfide-bridged PMO materials is retained, as that of the wheat-like SBA-15. Excellent adsorption efficiency (˜716 mg/g) for Hg 2+ ions is observed, suggesting a potential application in removal of heavy metal ions in wastewater.

  5. Lignin-assisted exfoliation of molybdenum disulfide in aqueous media and its application in lithium ion batteries.

    PubMed

    Liu, Wanshuang; Zhao, Chenyang; Zhou, Rui; Zhou, Dan; Liu, Zhaolin; Lu, Xuehong

    2015-06-01

    In this article, alkali lignin (AL)-assisted direct exfoliation of MoS2 mineral into single-layer and few-layer nanosheets in water is reported for the first time. Under optimized conditions, the concentration of MoS2 nanosheets in the obtained dispersion can be as high as 1.75 ± 0.08 mg mL(-1), which is much higher than the typical reported concentrations (<1.0 mg mL(-1)) using synthetic polymers or compounds as surfactants. The stabilizing mechanism primarily lies in the electrostatic repulsion between negative charged AL, as suggested by zeta-potential measurements. When the exfoliated MoS2 nanosheets are applied as electrode materials for lithium ion batteries, they show much improved electrochemical performance compared with the pristine MoS2 mineral because of the enhanced ion and electron transfer kinetics. This facile, scalable and eco-friendly aqueous-based process in combination with renewable and ultra-low-cost lignin opens up possibilities for large-scale fabrication of MoS2-based nanocomposites and devices. Moreover, herein we demonstrate that AL is also an excellent surfactant for exfoliation of many other types of layered materials, including graphene, tungsten disulfide and boron nitride, in water, providing rich opportunities for a wider range of applications. PMID:25970569

  6. Lignin-assisted exfoliation of molybdenum disulfide in aqueous media and its application in lithium ion batteries

    NASA Astrophysics Data System (ADS)

    Liu, Wanshuang; Zhao, Chenyang; Zhou, Rui; Zhou, Dan; Liu, Zhaolin; Lu, Xuehong

    2015-05-01

    In this article, alkali lignin (AL)-assisted direct exfoliation of MoS2 mineral into single-layer and few-layer nanosheets in water is reported for the first time. Under optimized conditions, the concentration of MoS2 nanosheets in the obtained dispersion can be as high as 1.75 +/- 0.08 mg mL-1, which is much higher than the typical reported concentrations (<1.0 mg mL-1) using synthetic polymers or compounds as surfactants. The stabilizing mechanism primarily lies in the electrostatic repulsion between negative charged AL, as suggested by zeta-potential measurements. When the exfoliated MoS2 nanosheets are applied as electrode materials for lithium ion batteries, they show much improved electrochemical performance compared with the pristine MoS2 mineral because of the enhanced ion and electron transfer kinetics. This facile, scalable and eco-friendly aqueous-based process in combination with renewable and ultra-low-cost lignin opens up possibilities for large-scale fabrication of MoS2-based nanocomposites and devices. Moreover, herein we demonstrate that AL is also an excellent surfactant for exfoliation of many other types of layered materials, including graphene, tungsten disulfide and boron nitride, in water, providing rich opportunities for a wider range of applications.In this article, alkali lignin (AL)-assisted direct exfoliation of MoS2 mineral into single-layer and few-layer nanosheets in water is reported for the first time. Under optimized conditions, the concentration of MoS2 nanosheets in the obtained dispersion can be as high as 1.75 +/- 0.08 mg mL-1, which is much higher than the typical reported concentrations (<1.0 mg mL-1) using synthetic polymers or compounds as surfactants. The stabilizing mechanism primarily lies in the electrostatic repulsion between negative charged AL, as suggested by zeta-potential measurements. When the exfoliated MoS2 nanosheets are applied as electrode materials for lithium ion batteries, they show much improved

  7. Carbon disulfide

    Integrated Risk Information System (IRIS)

    Carbon disulfide ; CASRN 75 - 15 - 0 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic E

  8. Exploiting Sequential Patterns Found in Users' Solutions and Virtual Tutor Behavior to Improve Assistance in ITS

    ERIC Educational Resources Information Center

    Fournier-Viger, Philippe; Faghihi, Usef; Nkambou, Roger; Nguifo, Engelbert Mephu

    2010-01-01

    We propose to mine temporal patterns in Intelligent Tutoring Systems (ITSs) to uncover useful knowledge that can enhance their ability to provide assistance. To discover patterns, we suggest using a custom, sequential pattern-mining algorithm. Two ways of applying the algorithm to enhance an ITS's capabilities are addressed. The first is to…

  9. Effect of deposition conditions on properties of plasma polymerized carbon disulfide

    SciTech Connect

    Sadhir, R.K.; Schoch, K.F. Jr.

    1995-12-31

    This paper discusses the results on deposition conditions, rates of polymerization and properties of carbon disulfide films prepared by two techniques viz. plasma polymerization and argon-plasma-assisted polymerization of carbon disulfide. A higher rate of polymerization and sulfur content was obtained for carbon disulfide films prepared by plasma polymerization technique. The ultimate objective of this research work was to prepare thin film solid state batteries using the optimized carbon disulfide polymer films deposited by plasma techniques, as active material.

  10. Microfiber-coupler-assisted control of wavelength tuning for Q-switched fiber laser with few-layer molybdenum disulfide nanoplates.

    PubMed

    Chen, Jin-hui; Deng, Guo-qing; Yan, Shao-cheng; Li, Cheng; Xi, Kai; Xu, Fei; Lu, Yan-qing

    2015-08-01

    Based on the liquid exfoliated method, we obtained the few-layer molybdenum disulfide (MoS2) nanoplates solution. By thermal evaporation method, we directly deposited MoS2 thin film onto the facet of a fiber patch cord. The modulation depth of the film is as high as 29%, and a Q-switched fiber laser was achieved. We also provided a new method to continuously tune the output laser with a tuning sensitivity of ∼5.5  nm/(1%  strain) by controlling the cavity loss with a strained microfiber coupler (MFC). PMID:26258361

  11. Cell-Penetrating Poly(disulfide) Assisted Intracellular Delivery of Mesoporous Silica Nanoparticles for Inhibition of miR-21 Function and Detection of Subsequent Therapeutic Effects.

    PubMed

    Yu, Changmin; Qian, Linghui; Ge, Jingyan; Fu, Jiaqi; Yuan, Peiyan; Yao, Samantha C L; Yao, Shao Q

    2016-08-01

    The design of drug delivery systems capable of minimal endolysosomal trapping, controlled drug release, and real-time monitoring of drug effect is highly desirable for personalized medicine. Herein, by using mesoporous silica nanoparticles (MSNs) coated with cell-penetrating poly(disulfide)s and a fluorogenic apoptosis-detecting peptide (DEVD-AAN), we have developed a platform that could be uptaken rapidly by mammalian cells via endocytosis-independent pathways. Subsequent loading of these MSNs with small molecule inhibitors and antisense oligonucleotides resulted in intracellular release of these drugs, leading to combination inhibition of endogenous miR-21 activities which was immediately detectable by the MSN surface-coated peptide using two-photon fluorescence microscopy. PMID:27325284

  12. Monolayer Tungsten Disulfide Laser

    NASA Astrophysics Data System (ADS)

    Ye, Yu; Wong, Zi Jing; Lu, Xiufang; Ni, Xingjie; Zhu, Hanyu; Chen, Xianhui; Wang, Yuan; Zhang, Xiang

    Two-dimensional van der Waals materials have opened a new paradigm for fundamental physics exploration and device applications because of their emerging physical properties. Unlike gapless graphene, monolayer transition-metal dichalcogenides are two-dimensional semiconductors that undergo an indirect-to-direct band gap transition, creating new optical functionalities for next-generation ultra-compact photonics and optoelectronics. Here, we report the realization of a two-dimensional excitonic laser by embedding monolayer tungsten disulfide in a microdisk resonator.

  13. Rapid sequencing and disulfide mapping of peptides containing disulfide bonds by using 1,5-diaminonaphthalene as a reductive matrix.

    PubMed

    Fukuyama, Yuko; Iwamoto, Shinichi; Tanaka, Koichi

    2006-02-01

    MS/MS is indispensable for the amino acid sequencing of peptides. However, its use is limited for peptides containing disulfide bonds. We have applied the reducing properties of 1,5-diaminonaphthalene (1,5-DAN) as a MALDI matrix to amino acid sequencing and disulfide bond mapping of human urotensin II possessing one disulfide bond, and human guanylin possessing two disulfide bonds. 1,5-DAN was used in the same manner as the usual MALDI matrices without any pre-treatment of the peptide, and MS/MS was performed using a matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer (MALDI QIT TOFMS). The results demonstrated that MS/MS of the molecular ions reduced by 1,5-DAN provided a series of significant b-/y-product ions. All 11 amino acid residues of urotensin II were identified using 1,5-DAN, while only 5 out of 11 residues were identified using 2,5-dihydroxybenzoic acid (DHB); similarly 11 out of 15 amino acid residues of guanylin were identified using 1,5-DAN, while only three were identified using DHB. In addition, comparison of the theoretical and measured values of the mass differences between corresponding MS/MS product ions using 1,5-DAN and DHB narrowed down the possible disulfide bond arrangement candidates. Consequently, 1,5-DAN as a reductive matrix facilitates rapid amino acid sequencing and disulfide mapping for peptides containing disulfide bonds. PMID:16382486

  14. 46 CFR 153.1040 - Carbon disulfide.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Carbon disulfide. 153.1040 Section 153.1040 Shipping... § 153.1040 Carbon disulfide. (a) No person may load, carry, or discharge carbon disulfide unless the... charge of a carbon disulfide transfer operation shall ensure that carbon disulfide is discharged only...

  15. 46 CFR 153.1040 - Carbon disulfide.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Carbon disulfide. 153.1040 Section 153.1040 Shipping... § 153.1040 Carbon disulfide. (a) No person may load, carry, or discharge carbon disulfide unless the... charge of a carbon disulfide transfer operation shall ensure that carbon disulfide is discharged only...

  16. 46 CFR 153.1040 - Carbon disulfide.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Carbon disulfide. 153.1040 Section 153.1040 Shipping... § 153.1040 Carbon disulfide. (a) No person may load, carry, or discharge carbon disulfide unless the... charge of a carbon disulfide transfer operation shall ensure that carbon disulfide is discharged only...

  17. 46 CFR 153.1040 - Carbon disulfide.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Carbon disulfide. 153.1040 Section 153.1040 Shipping... § 153.1040 Carbon disulfide. (a) No person may load, carry, or discharge carbon disulfide unless the... charge of a carbon disulfide transfer operation shall ensure that carbon disulfide is discharged only...

  18. 46 CFR 153.1040 - Carbon disulfide.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Carbon disulfide. 153.1040 Section 153.1040 Shipping... § 153.1040 Carbon disulfide. (a) No person may load, carry, or discharge carbon disulfide unless the... charge of a carbon disulfide transfer operation shall ensure that carbon disulfide is discharged only...

  19. Quantification of Thiols and Disulfides

    PubMed Central

    Winther, Jakob R.; Thorpe, Colin

    2013-01-01

    Background Disulfide bond formation is a key posttranslational modification, with implications for structure, function and stability of numerous proteins. While disulfide bond formation is a necessary and essential process for many proteins, it is deleterious and disruptive for others. Cells go to great lengths to regulate thiol-disulfide bond homeostasis, typically with several, apparently redundant, systems working in parallel. Dissecting the extent of oxidation and reduction of disulfides is an ongoing challenge due, in part, to the facility of thiol/disulfide exchange reactions. Scope of the review In the present account, we briefly survey the toolbox available to the experimentalist for the chemical determination of thiols and disulfides. We have chosen to focus on the key chemical aspects of current methodology, together with identifying potential difficulties inherent in their experimental implementation. Major conclusions While many reagents have been described for the measurement and manipulation of the redox status of thiols and disulfides, a number of these methods remain underutilized. The ability to effectively quantify changes in redox conditions in living cells presents a continuing challenge. General Significance Many unresolved questions in the metabolic interconversion of thiols and disulfides remain. For example, while pool sizes of redox pairs and their intracellular distribution are being uncovered, very little is known about the flux in thiol-disulfide exchange pathways. New tools are needed to address this important aspect of cellular metabolism. PMID:23567800

  20. Twin disulfides for orthogonal disulfide pairing and the directed folding of multicyclic peptides

    NASA Astrophysics Data System (ADS)

    Wu, Chuanliu; Leroux, Jean-Christophe; Gauthier, Marc A.

    2012-12-01

    Multicyclic peptides are emerging as an exciting platform for drug and targeted ligand discovery owing to their expected greater target affinity/selectivity/stability versus linear or monocyclic peptides. However, although the precise pairing of cysteine residues in proteins is routinely achieved in nature, the rational pairing of cysteine residues within polypeptides is a long-standing challenge for the preparation of multicyclic species containing several disulfide bridges. Here, we present an efficient and straightforward approach for directing the intermolecular and intramolecular pairing of cysteine residues within peptides using a minimal CXC motif. Orthogonal disulfide pairing can be exploited in complex redox media to rationally produce dimeric peptides and bi/tricyclic peptides from fully reduced peptides containing 1-6 cysteine residues. This strategy, which does not rely on extensive manipulation of the primary sequence, post-translational modification or protecting groups, should greatly benefit the development of multicyclic peptide therapeutics and targeting ligands.

  1. Dynamic Chemistry of Disulfide Terminated Oligonucleotides in Duplexes and Double-Crossover Tiles.

    PubMed

    De Stefano, Mattia; Vesterager Gothelf, Kurt

    2016-06-16

    Designed nanostructures formed by self-assembly of multiple DNA strands suffer from low stability at elevated temperature and under other denaturing conditions. Here, we propose a method for covalent coupling of DNA strands in such structures by the formation of disulfide bonds; this allows disassembly of the structure under reducing conditions. The dynamic chemistry of disulfides and thiols was applied to crosslink DNA strands with terminal disulfide modifications. The formation of disulfide-linked DNA duplexes consisting of three strands is demonstrated, as well as a more-complex DNA double-crossover tile. All the strands in the fully disulfide-linked structures are covalently and geometrically interlocked, and it is demonstrated that the structures are stable under heating and in the presence of denaturants. Such a reversible system can be exploited in applications where higher DNA stability is needed only temporarily, such as delivery of cargoes to cells by DNA nanostructures. PMID:26994867

  2. Thermodynamic properties of zirconium disulfide

    SciTech Connect

    Volovik, L.S.; Kovalevskaya, E.I.; Litvinenko, V.F.

    1986-02-01

    This paper uses a method of comparative calculation -- double comparison -for the quantitative evaluation of the themodynamic characteristics of zirconium disulfide. The method enables one to apply known characteristics of compounds of the given or an adjacent group to analogous compounds of elements. The enthalpy of zirconium disulfide was determined by a formula and the calculation was carried out on the basis of data on the enthalpy of hafnium, niobium, and tantalum disulfides measured by the mixing method in the temperature range 500-1800 K.

  3. Plasma-polymerized carbon disulfide thin-film rechargeable batteries

    SciTech Connect

    Sadhir, R.K.; Schoch, K.F. Jr.

    1996-06-01

    This paper discusses the results on deposition conditions, rates of polymerization, and properties of carbon disulfide films prepared by two techniques, viz., plasma polymerization and argon-plasma-assisted polymerization of carbon disulfide. A higher rate of polymerization and sulfur content was obtained for carbon disulfide films prepared by the plasma-polymerization technique. A Li/LiClO{sub 4} propylene carbonate/PPCS cell was assembled using lithium foil, 2 M LiClO{sub 4}/PC electrolyte with polyolefin separator, and PPCS deposited on Pt foil. The electrode area was 1.3 cm{sup 2}. The cell cycled reversibly between 1 and 3 V and retained its capacity (5 mA h/g) well over 12 cycles. 14 refs., 11 figs., 5 tabs.

  4. Induction and identification of disulfide-intact and disulfide-reduced beta-subunit of Shiga toxin 2 from Escherichia coli O157:H7 using MALDI-TOF-TOF-MS/MS and top-down proteomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The disulfide-intact and disulfide-reduced beta-subunit of Shiga toxin 2 (beta-Stx2) from Escherichia coli O157:H7 (strain EDL933) has been identified by matrix-assisted laser desorption/ionization time-of-flight-time-of-flight tandem mass spectrometry (MALDI-TOF-TOF-MS/MS) and top-down proteomic an...

  5. 40 CFR 721.1745 - Ethoxybenzothiazole disulfide.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Ethoxybenzothiazole disulfide. 721... Substances § 721.1745 Ethoxybenzothiazole disulfide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance generically identified as ethoxybenzothiazole disulfide...

  6. 40 CFR 721.1745 - Ethoxybenzothiazole disulfide.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Ethoxybenzothiazole disulfide. 721... Substances § 721.1745 Ethoxybenzothiazole disulfide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance generically identified as ethoxybenzothiazole disulfide...

  7. 40 CFR 721.1745 - Ethoxybenzothiazole disulfide.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Ethoxybenzothiazole disulfide. 721... Substances § 721.1745 Ethoxybenzothiazole disulfide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance generically identified as ethoxybenzothiazole disulfide...

  8. 40 CFR 721.1745 - Ethoxybenzothiazole disulfide.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Ethoxybenzothiazole disulfide. 721... Substances § 721.1745 Ethoxybenzothiazole disulfide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance generically identified as ethoxybenzothiazole disulfide...

  9. Orthogonal Cysteine-Penicillamine Disulfide Pairing for Directing the Oxidative Folding of Peptides.

    PubMed

    Zheng, Yiwu; Zhai, Linxiang; Zhao, Yibing; Wu, Chuanliu

    2015-12-01

    Precise disulfide pairing in synthetic peptides usually is achieved using orthogonal protecting group strategies or relies on primary sequence manipulation. Orthogonal disulfide pairing technology should be promising for directing the rational folding of multicyclic peptides from the fully reduced peptides. Here, we report a discovery on the orthogonality between heterodisulfide pairing of cysteine (Cys) and penicillamine (Pen) and formation of Cys-Cys/Pen-Pen homodisulfides. The orthogonal Cys-Pen disulfide pairing can be exploited for highly selective production of certain (multi)cyclic structures (or even a sole structure without isomers) through direct oxidation in air or thiol-disulfide exchanges in redox media. This strategy makes rational folding of multicyclic peptides without protecting groups, sequence manipulation, and complex synthetic reactions a reality, thus providing invaluable assets to peptide communities, and should greatly benefit the development of multicyclic peptide therapeutics and ligands. PMID:26588670

  10. Intradomain Confinement of Disulfides in the Folding of Two Consecutive Modules of the LDL Receptor

    PubMed Central

    Martínez-Oliván, Juan; Fraga, Hugo; Arias-Moreno, Xabier; Ventura, Salvador; Sancho, Javier

    2015-01-01

    The LDL receptor internalizes circulating LDL and VLDL particles for degradation. Its extracellular binding domain contains ten (seven LA and three EGF) cysteine-rich modules, each bearing three disulfide bonds. Despite the enormous number of disulfide combinations possible, LDLR oxidative folding leads to a single native species with 30 unique intradomain disulfides. Previous folding studies of the LDLR have shown that non native disulfides are initially formed that lead to compact species. Accordingly, the folding of the LDLR has been described as a "coordinated nonvectorial” reaction, and it has been proposed that early compaction funnels the reaction toward the native structure. Here we analyze the oxidative folding of LA4 and LA5, the modules critical for ApoE binding, isolated and in the LA45 tandem. Compared to LA5, LA4 folding is slow and inefficient, resembling that of LA5 disease-linked mutants. Without Ca++, it leads to a mixture of many two-disulfide scrambled species and, with Ca++, to the native form plus two three-disulfide intermediates. The folding of the LA45 tandem seems to recapitulate that of the individual repeats. Importantly, although the folding of the LA45 tandem takes place through formation of scrambled isomers, no interdomain disulfides are detected, i.e. the two adjacent modules fold independently without the assistance of interdomain covalent interactions. Reduction of incredibly large disulfide combinatorial spaces, such as that in the LDLR, by intradomain confinement of disulfide bond formation might be also essential for the efficient folding of other homologous disulfide-rich receptors. PMID:26168158

  11. In-Depth Characterization of Protein Disulfide Bonds by Online Liquid Chromatography-Electrochemistry-Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Switzar, Linda; Nicolardi, Simone; Rutten, Julie W.; Oberstein, Saskia A. J. Lesnik; Aartsma-Rus, Annemieke; van der Burgt, Yuri E. M.

    2016-01-01

    Disulfide bonds are an important class of protein post-translational modifications, yet this structurally crucial modification type is commonly overlooked in mass spectrometry (MS)-based proteomics approaches. Recently, the benefits of online electrochemistry-assisted reduction of protein S-S bonds prior to MS analysis were exemplified by successful characterization of disulfide bonds in peptides and small proteins. In the current study, we have combined liquid chromatography (LC) with electrochemistry (EC) and mass analysis by Fourier transform ion cyclotron resonance (FTICR) MS in an online LC-EC-MS platform to characterize protein disulfide bonds in a bottom-up proteomics workflow. A key advantage of a LC-based strategy is the use of the retention time in identifying both intra- and interpeptide disulfide bonds. This is demonstrated by performing two sequential analyses of a certain protein digest, once without and once with electrochemical reduction. In this way, the "parent" disulfide-linked peptide detected in the first run has a retention time-based correlation with the EC-reduced peptides detected in the second run, thus simplifying disulfide bond mapping. Using this platform, both inter- and intra-disulfide-linked peptides were characterized in two different proteins, ß-lactoglobulin and ribonuclease B. In order to prevent disulfide reshuffling during the digestion process, proteins were digested at a relatively low pH, using (a combination of) the high specificity proteases trypsin and Glu-C. With this approach, disulfide bonds in ß-lactoglobulin and ribonuclease B were comprehensively identified and localized, showing that online LC-EC-MS is a useful tool for the characterization of protein disulfide bonds.

  12. Disulfide Trapping for Modeling and Structure Determination of Receptor:Chemokine Complexes

    PubMed Central

    Kufareva, Irina; Gustavsson, Martin; Holden, Lauren G.; Qin, Ling; Zheng, Yi; Handel, Tracy M.

    2016-01-01

    Despite the recent breakthrough advances in GPCR crystallography, structure determination of protein-protein complexes involving chemokine receptors and their endogenous chemokine ligands remains challenging. Here we describe disulfide trapping, a methodology for generating irreversible covalent binary protein complexes from unbound protein partners by introducing two cysteine residues, one per interaction partner, at selected positions within their interaction interface. Disulfide trapping can serve at least two distinct purposes: (i) stabilization of the complex to assist structural studies, and/or (ii) determination of pairwise residue proximities to guide molecular modeling. Methods for characterization of disulfide-trapped complexes are described and evaluated in terms of throughput, sensitivity, and specificity towards the most energetically favorable cross-links. Due to abundance of native disulfide bonds at receptor:chemokine interfaces, disulfide trapping of their complexes can be associated with intramolecular disulfide shuffling and result in misfolding of the component proteins; because of this, evidence from several experiments is typically needed to firmly establish a positive disulfide crosslink. An optimal pipeline that maximizes throughput and minimizes time and costs by early triage of unsuccessful candidate constructs is proposed. PMID:26921956

  13. Mixed results with mixed disulfides.

    PubMed

    Brigelius-Flohé, Regina

    2016-04-01

    A period of research with Helmut Sies in the 1980s is recalled. Our experiments aimed at an in-depth understanding of metabolic changes due to oxidative challenges under near-physiological conditions, i.e. perfused organs. A major focus were alterations of the glutathione and the NADPH/NADP(+) system by different kinds of oxidants, in particular formation of glutathione mixed disulfides with proteins. To analyze mixed disulfides, a test was adapted which is widely used until today. The observations in perfused rat livers let us believe that glutathione-6-phosphate dehydrogenase (G6PDH), i.a. might be activated by glutathionylation. Although we did not succeed to verify this hypothesis for the special case of G6PDH, the regulation of enzyme/protein activities by glutathionylation today is an accepted posttranslational mechanism in redox biology in general. Our early experimental approaches are discussed in the context of present knowledge. PMID:27095221

  14. Molybdenum disulfide (MoS2) nanoflakes as inherently electroactive labels for DNA hybridization detection.

    PubMed

    Loo, Adeline Huiling; Bonanni, Alessandra; Ambrosi, Adriano; Pumera, Martin

    2014-10-21

    The detection of specific DNA sequences plays a critical role in the areas of medical diagnostics, environmental monitoring, drug discovery and food safety. This has therefore become a strong driving force behind the ever-increasing demand for simple, cost-effective, highly sensitive and selective DNA biosensors. In this study, we report for the first time, a novel approach for the utilization of molybdenum disulfide nanoflakes, a member of the transition metal dichalcogenides family, in the detection of DNA hybridization. Herein, molybdenum disulfide nanoflakes serve as inherently electroactive labels, with the inherent oxidation peak exploited as the analytical signal. The principle of detection is based on the differential affinity of molybdenum disulfide nanoflakes towards single-stranded DNA and double-stranded DNA. The employment of transition metal dichalcogenide nanomaterials for sensing and biosensing purposes represents an upcoming research area which holds great promise. Hence, our findings are anticipated to have significant contributions towards the fabrication of future DNA biosensors. PMID:25177907

  15. Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli

    PubMed Central

    de Marco, Ario

    2009-01-01

    Bacteria are simple and cost effective hosts for producing recombinant proteins. However, their physiological features may limit their use for obtaining in native form proteins of some specific structural classes, such as for instance polypeptides that undergo extensive post-translational modifications. To some extent, also the production of proteins that depending on disulfide bridges for their stability has been considered difficult in E. coli. Both eukaryotic and prokaryotic organisms keep their cytoplasm reduced and, consequently, disulfide bond formation is impaired in this subcellular compartment. Disulfide bridges can stabilize protein structure and are often present in high abundance in secreted proteins. In eukaryotic cells such bonds are formed in the oxidizing environment of endoplasmic reticulum during the export process. Bacteria do not possess a similar specialized subcellular compartment, but they have both export systems and enzymatic activities aimed at the formation and at the quality control of disulfide bonds in the oxidizing periplasm. This article reviews the available strategies for exploiting the physiological mechanisms of bactera to produce properly folded disulfide-bonded proteins. PMID:19442264

  16. Transfer of molybdenum disulfide to various metals

    NASA Technical Reports Server (NTRS)

    Barton, G. C.; Pepper, S. V.

    1977-01-01

    Sliding friction experiments were conducted with molybdenum disulfide single crystals in contact with sputter cleaned surfaces of copper, nickel, gold, and 304 stainless steel. Transfer of the molybdenum disulfide to the metals was monitored with Auger electron spectroscopy. Results of the investigation indicate molybdenum disulfide transfers to all clean metal surfaces after a single pass over the metal surface with film thickness observed to increase with repeated passes over the same surfaces. Large particle transfer occurs when the orientation of the crystallites is other than basal. This is frequently accompanied by abrasion of the metal. Adhesion of molybdenum disulfide films occurred readily to copper and nickel, less readily to 304 stainless steel, and even less effectively to the gold, which indicates a chemical effect.

  17. Protein folding guides disulfide bond formation

    PubMed Central

    Qin, Meng; Wang, Wei; Thirumalai, D.

    2015-01-01

    The Anfinsen principle that the protein sequence uniquely determines its structure is based on experiments on oxidative refolding of a protein with disulfide bonds. The problem of how protein folding drives disulfide bond formation is poorly understood. Here, we have solved this long-standing problem by creating a general method for implementing the chemistry of disulfide bond formation and rupture in coarse-grained molecular simulations. As a case study, we investigate the oxidative folding of bovine pancreatic trypsin inhibitor (BPTI). After confirming the experimental findings that the multiple routes to the folded state contain a network of states dominated by native disulfides, we show that the entropically unfavorable native single disulfide [14–38] between Cys14 and Cys38 forms only after polypeptide chain collapse and complete structuring of the central core of the protein containing an antiparallel β-sheet. Subsequent assembly, resulting in native two-disulfide bonds and the folded state, involves substantial unfolding of the protein and transient population of nonnative structures. The rate of [14–38] formation increases as the β-sheet stability increases. The flux to the native state, through a network of kinetically connected native-like intermediates, changes dramatically by altering the redox conditions. Disulfide bond formation between Cys residues not present in the native state are relevant only on the time scale of collapse of BPTI. The finding that formation of specific collapsed native-like structures guides efficient folding is applicable to a broad class of single-domain proteins, including enzyme-catalyzed disulfide proteins. PMID:26297249

  18. The Indian Athlete: Exploiting or Exploited?

    ERIC Educational Resources Information Center

    Salter, Michael A.

    It is the purpose of this paper to examine the nineteenth century Canadian Indian lacrosse player to determine whether or not he was exploited by his European counterparts, and if so, the manner in which this exploitation occurred. Caucasian lacrosse enthusiasts sought to promote "their" game by arranging for Indian demonstrations to be staged…

  19. Disulfide bond assignments by mass spectrometry of native natural peptides: cysteine pairing in disulfide bonded conotoxins.

    PubMed

    Gupta, Kallol; Kumar, Mukesh; Balaram, Padmanabhan

    2010-10-01

    The critical, and often most difficult, step in structure elucidation of diverse classes of natural peptides is the determination of correct disulfide pairing between multiple cysteine residues. Here, we present a direct mass spectrometric analytical methodology for the determination of disulfide pairing. Protonated peptides, having multiple disulfide bonds, fragmented under collision induced dissociation (CID) conditions and preferentially cleave along the peptide backbone, with occasional disulfide fragmentation either by C(β)-S bond cleavage through H(α) abstraction to yield dehydroalanine and cysteinepersulfide, or by S-S bond cleavage through H(β) abstraction to yield the thioaldehyde and cysteine. Further fragmentation of the initial set of product ions (MS(n)) yields third and fourth generation fragment ions, permitting a distinction between the various possible disulfide bonded structures. This approach is illustrated by establishing cysteine pairing patterns in five conotoxins containing two disulfide bonds. The methodology is extended to the Conus araneosus peptides Ar1446 and Ar1430, two 14 residue sequences containing 3 disulfide bonds. A distinction between 15 possible disulfide pairing schemes becomes possible using direct mass spectral fragmentation of the native peptides together with fragmentation of enzymatically nicked peptides. PMID:20843009

  20. How thioredoxin dissociates its mixed disulfide.

    PubMed

    Roos, Goedele; Foloppe, Nicolas; Van Laer, Koen; Wyns, Lode; Nilsson, Lennart; Geerlings, Paul; Messens, Joris

    2009-08-01

    The dissociation mechanism of the thioredoxin (Trx) mixed disulfide complexes is unknown and has been debated for more than twenty years. Specifically, opposing arguments for the activation of the nucleophilic cysteine as a thiolate during the dissociation of the complex have been put forward. As a key model, the complex between Trx and its endogenous substrate, arsenate reductase (ArsC), was used. In this structure, a Cys29(Trx)-Cys89(ArsC) intermediate disulfide is formed by the nucleophilic attack of Cys29(Trx) on the exposed Cys82(ArsC)-Cys89(ArsC) in oxidized ArsC. With theoretical reactivity analysis, molecular dynamics simulations, and biochemical complex formation experiments with Cys-mutants, Trx mixed disulfide dissociation was studied. We observed that the conformational changes around the intermediate disulfide bring Cys32(Trx) in contact with Cys29(Trx). Cys32(Trx) is activated for its nucleophilic attack by hydrogen bonds, and Cys32(Trx) is found to be more reactive than Cys82(ArsC). Additionally, Cys32(Trx) directs its nucleophilic attack on the more susceptible Cys29(Trx) and not on Cys89(ArsC). This multidisciplinary approach provides fresh insights into a universal thiol/disulfide exchange reaction mechanism that results in reduced substrate and oxidized Trx. PMID:19675666

  1. Battlespace exploitation vision

    NASA Astrophysics Data System (ADS)

    Gilmore, John F.; Johnson, Ray O.

    1997-06-01

    The quantity and quality of data collected by military and commercial electro-optic and radar sensors is rapidly increasing. This increase in imagery data has not been accompanied by an increase in the number of image analysts needed to rapidly screen the imagery to locate and identify military targets or other objects of interest. Automatic target recognition (ATR) technology that automates the target detection, classification, and identification process has been a promising technology for at least two decades, and recent advances make the realization of aided target recognition possible. The future military battlespace will be filled with airborne, spaceborne, and land-based sensors observing moving and stationary targets at various locations, from multiple aspects, and at multiple frequencies and wavelengths. Only through the use of computer-assisted data analysis and ATR can the vast amount of data be analyzed within the timelines required by the military. The Defense Advanced Research Projects Agency (DARPA) has a number of programs developing technology to support the exploitation and control of the future battlespace information.

  2. Identification of protein disulfide isomerase 1 as a key isomerase for disulfide bond formation in apolipoprotein B100.

    PubMed

    Wang, Shiyu; Park, Shuin; Kodali, Vamsi K; Han, Jaeseok; Yip, Theresa; Chen, Zhouji; Davidson, Nicholas O; Kaufman, Randal J

    2015-02-15

    Apolipoprotein (apo) B is an obligatory component of very low density lipoprotein (VLDL), and its cotranslational and posttranslational modifications are important in VLDL synthesis, secretion, and hepatic lipid homeostasis. ApoB100 contains 25 cysteine residues and eight disulfide bonds. Although these disulfide bonds were suggested to be important in maintaining apoB100 function, neither the specific oxidoreductase involved nor the direct role of these disulfide bonds in apoB100-lipidation is known. Here we used RNA knockdown to evaluate both MTP-dependent and -independent roles of PDI1 in apoB100 synthesis and lipidation in McA-RH7777 cells. Pdi1 knockdown did not elicit any discernible detrimental effect under normal, unstressed conditions. However, it decreased apoB100 synthesis with attenuated MTP activity, delayed apoB100 oxidative folding, and reduced apoB100 lipidation, leading to defective VLDL secretion. The oxidative folding-impaired apoB100 was secreted mainly associated with LDL instead of VLDL particles from PDI1-deficient cells, a phenotype that was fully rescued by overexpression of wild-type but not a catalytically inactive PDI1 that fully restored MTP activity. Further, we demonstrate that PDI1 directly interacts with apoB100 via its redox-active CXXC motifs and assists in the oxidative folding of apoB100. Taken together, these findings reveal an unsuspected, yet key role for PDI1 in oxidative folding of apoB100 and VLDL assembly. PMID:25518935

  3. Reactive sputter deposition of pyrite structure transition metal disulfide thin films: Microstructure, transport, and magnetism

    SciTech Connect

    Baruth, A.; Manno, M.; Narasimhan, D.; Shankar, A.; Zhang, X.; Johnson, M.; Aydil, E. S.; Leighton, C.

    2012-09-01

    Transition metal disulfides crystallizing in the pyrite structure (e.g., TMS{sub 2}, with TM = Fe, Co, Ni, and Cu) are a class of materials that display a remarkably diverse array of functional properties. These properties include highly spin-polarized ferromagnetism (in Co{sub 1-x}Fe{sub x}S{sub 2}), superconductivity (in CuS{sub 2}), an antiferromagnetic Mott insulating ground state (in NiS{sub 2}), and semiconduction with close to optimal parameters for solar absorber applications (in FeS{sub 2}). Exploitation of these properties in heterostructured devices requires the development of reliable and reproducible methods for the deposition of high quality pyrite structure thin films. In this manuscript, we report on the suitability of reactive sputter deposition from metallic targets in an Ar/H{sub 2}S environment as a method to achieve exactly this. Optimization of deposition temperature, Ar/H{sub 2}S pressure ratio, and total working gas pressure, assisted by plasma optical emission spectroscopy, reveals significant windows over which deposition of single-phase, polycrystalline, low roughness pyrite films can be achieved. This is illustrated for the test cases of the ferromagnetic metal CoS{sub 2} and the diamagnetic semiconductor FeS{sub 2}, for which detailed magnetic and transport characterization are provided. The results indicate significant improvements over alternative deposition techniques such as ex situ sulfidation of metal films, opening up exciting possibilities for all-sulfide heterostructured devices. In particular, in the FeS{sub 2} case it is suggested that fine-tuning of the sputtering conditions provides a potential means to manipulate doping levels and conduction mechanisms, critical issues in solar cell applications. Parenthetically, we note that conditions for synthesis of phase-pure monosulfides and thiospinels are also identified.

  4. Disulfide-Functionalized Diblock Copolymer Worm Gels.

    PubMed

    Warren, Nicholas J; Rosselgong, Julien; Madsen, Jeppe; Armes, Steven P

    2015-08-10

    Two strategies for introducing disulfide groups at the outer surface of RAFT-synthesized poly(glycerol monomethacrylate)-poly(2-hydroxypropyl methacrylate) (PGMA-PHPMA, or Gx-Hy for brevity) diblock copolymer worms are investigated. The first approach involved statistical copolymerization of GMA with a small amount of disulfide dimethacrylate (DSDMA, or D) comonomer to afford a G54-D0.50 macromolecular chain transfer agent (macro-CTA); this synthesis was conducted in relatively dilute solution in order to ensure mainly intramolecular cyclization and hence the formation of linear chains. Alternatively, a new disulfide-based bifunctional RAFT agent (DSDB) was used to prepare a G45-S-S-G45 (or (G45-S)2) macro-CTA. A binary mixture of a non-functionalized G55 macro-CTA was utilized with each of these two disulfide-based macro-CTAs in turn for the RAFT aqueous dispersion polymerization of 2-hydroxypropyl methacrylate (HPMA). By targeting a PHPMA DP of 130 and systematically varying the molar ratio of the two macro-CTAs, a series of disulfide-functionalized diblock copolymer worm gels were obtained. For both formulations, oscillatory rheology studies confirmed that higher disulfide contents led to stronger gels, presumably as a result of inter-worm covalent bond formation via disulfide/thiol exchange. Using the DSDB-based macro-CTA led to the strongest worm gels, and this formulation also proved to be more effective in suppressing the thermosensitive behavior that is observed for the nondisulfide-functionalized control worm gel. However, macroscopic precipitation occurred when the proportion of DSDB-based macro-CTA was increased to 50 mol %, whereas the DSDMA-based macro-CTA could be utilized at up to 80 mol %. Finally, the worm gel modulus could be reduced to that of a nondisulfide-containing worm gel by reductive cleavage of the inter-worm disulfide bonds using excess tris(2-carboxyethyl)phosphine (TCEP) to yield thiol groups. These new biomimetic worm gels are

  5. Voltammetric studies of poly(carbon disulfide)

    SciTech Connect

    Geng, L.; Xu, J.; Prasad, S.; Skotheim, T.A.; Lee, H.S.; McBreen, J.

    1992-12-31

    Poly(carbon disulfide) was studied by cyclic voltammetry using glassy carbon and platinum macro- and microdisk electrodes. The electron transfer kinetics is significantly faster at glassy carbon electrodes than at Pt electrodes. It is chemically reversible with moderate electron transfer rates. Voltammetric results of poly(carbon disulfide) are in good agreement with battery testing data. The k{sup 0} value measured at a Pt microdisk electrode is 7{times}10{sup 3} cm/sec. Electrochemical data suggest that PCS can be a potential cathode material for low current density lithium batteries.

  6. Selective and efficient electrochemical biosensing of ultrathin molybdenum disulfide sheets

    NASA Astrophysics Data System (ADS)

    Narayanan, Tharangattu N.; Vusa, Chiranjeevi S. R.; Alwarappan, Subbiah

    2014-08-01

    Atomically thin molybdenum disulfide (MoS2) sheets were synthesized and isolated via solvent-assisted chemical exfoliation. The charge-dependent electrochemical activities of these MoS2 sheets were studied using positively charged hexamine ruthenium (III) chloride and negatively charged ferricyanide/ferrocyanide redox probes. Ultrathin MoS2 sheet-based electrodes were employed for the electrochemical detection of an important neurotransmitter, namely dopamine (DA), in the presence of ascorbic acid (AA). MoS2 electrodes were identified as being capable of distinguishing the coexistence of the DA and the AA with an excellent stability. Moreover, the enzymatic detection of the glucose was studied by immobilizing glucose oxidase on the MoS2. This study opens enzymatic and non-enzymatic electrochemical biosensing applications of atomic MoS2 sheets, which will supplement their established electronic applications.

  7. DISULFIND: a disulfide bonding state and cysteine connectivity prediction server

    PubMed Central

    Ceroni, Alessio; Passerini, Andrea; Vullo, Alessandro; Frasconi, Paolo

    2006-01-01

    DISULFIND is a server for predicting the disulfide bonding state of cysteines and their disulfide connectivity starting from sequence alone. Optionally, disulfide connectivity can be predicted from sequence and a bonding state assignment given as input. The output is a simple visualization of the assigned bonding state (with confidence degrees) and the most likely connectivity patterns. The server is available at . PMID:16844986

  8. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Carbon disulfide; tolerances for... § 180.467 Carbon disulfide; tolerances for residues. Tolerances are established for the nematicide, insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  9. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Carbon disulfide; tolerances for... § 180.467 Carbon disulfide; tolerances for residues. Tolerances are established for the nematicide, insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  10. 46 CFR 153.520 - Special requirements for carbon disulfide.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Special requirements for carbon disulfide. 153.520... Equipment Special Requirements § 153.520 Special requirements for carbon disulfide. A containment system carrying carbon disulfide must meet the following: (a) Each cargo pump must be of the intank type...

  11. 46 CFR 153.520 - Special requirements for carbon disulfide.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Special requirements for carbon disulfide. 153.520... Equipment Special Requirements § 153.520 Special requirements for carbon disulfide. A containment system carrying carbon disulfide must meet the following: (a) Each cargo pump must be of the intank type...

  12. 46 CFR 153.520 - Special requirements for carbon disulfide.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Special requirements for carbon disulfide. 153.520... Equipment Special Requirements § 153.520 Special requirements for carbon disulfide. A containment system carrying carbon disulfide must meet the following: (a) Each cargo pump must be of the intank type...

  13. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Carbon disulfide; tolerances for... § 180.467 Carbon disulfide; tolerances for residues. Tolerances are established for the nematicide, insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  14. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Carbon disulfide; tolerances for... § 180.467 Carbon disulfide; tolerances for residues. Tolerances are established for the nematicide, insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  15. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Carbon disulfide; tolerances for... § 180.467 Carbon disulfide; tolerances for residues. Tolerances are established for the nematicide, insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  16. 46 CFR 153.520 - Special requirements for carbon disulfide.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Special requirements for carbon disulfide. 153.520... Equipment Special Requirements § 153.520 Special requirements for carbon disulfide. A containment system carrying carbon disulfide must meet the following: (a) Each cargo pump must be of the intank type...

  17. 46 CFR 153.520 - Special requirements for carbon disulfide.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Special requirements for carbon disulfide. 153.520... Equipment Special Requirements § 153.520 Special requirements for carbon disulfide. A containment system carrying carbon disulfide must meet the following: (a) Each cargo pump must be of the intank type...

  18. 21 CFR 524.2101 - Selenium disulfide suspension.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Selenium disulfide suspension. 524.2101 Section... § 524.2101 Selenium disulfide suspension. (a) Specifications. The product contains 0.9-percent weight in weight (w/w) selenium disulfide (1-percent weight in volume (w/v)). (b) Sponsors. See Nos. 000061,...

  19. 21 CFR 524.2101 - Selenium disulfide suspension.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Selenium disulfide suspension. 524.2101 Section... § 524.2101 Selenium disulfide suspension. (a) Specifications. The product contains 0.9-percent weight in weight (w/w) selenium disulfide (1-percent weight in volume (w/v)). (b) Sponsors. See Nos. 000061,...

  20. 21 CFR 524.2101 - Selenium disulfide suspension.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Selenium disulfide suspension. 524.2101 Section... § 524.2101 Selenium disulfide suspension. (a) Specifications. The product contains 0.9-percent weight in weight (w/w) selenium disulfide (1-percent weight in volume (w/v)). (b) Sponsors. See Nos. 000061,...

  1. 21 CFR 524.2101 - Selenium disulfide suspension.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Selenium disulfide suspension. 524.2101 Section... § 524.2101 Selenium disulfide suspension. (a) Specifications. The product contains 0.9-percent weight in weight (w/w) selenium disulfide (1-percent weight in volume (w/v)). (b) Sponsors. See Nos. 000061,...

  2. 21 CFR 524.2101 - Selenium disulfide suspension.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Selenium disulfide suspension. 524.2101 Section... § 524.2101 Selenium disulfide suspension. (a) Specifications. The product contains 0.9-percent weight in weight (w/w) selenium disulfide (1-percent weight in volume (w/v)). (b) Sponsors. See Nos. 000061,...

  3. Enhancing protein stability with extended disulfide bonds.

    PubMed

    Liu, Tao; Wang, Yan; Luo, Xiaozhou; Li, Jack; Reed, Sean A; Xiao, Han; Young, Travis S; Schultz, Peter G

    2016-05-24

    Disulfide bonds play an important role in protein folding and stability. However, the cross-linking of sites within proteins by cysteine disulfides has significant distance and dihedral angle constraints. Here we report the genetic encoding of noncanonical amino acids containing long side-chain thiols that are readily incorporated into both bacterial and mammalian proteins in good yields and with excellent fidelity. These amino acids can pair with cysteines to afford extended disulfide bonds and allow cross-linking of more distant sites and distinct domains of proteins. To demonstrate this notion, we preformed growth-based selection experiments at nonpermissive temperatures using a library of random β-lactamase mutants containing these noncanonical amino acids. A mutant enzyme that is cross-linked by one such extended disulfide bond and is stabilized by ∼9 °C was identified. This result indicates that an expanded set of building blocks beyond the canonical 20 amino acids can lead to proteins with improved properties by unique mechanisms, distinct from those possible through conventional mutagenesis schemes. PMID:27162342

  4. Preliminary Hazards Assessment: Iron disulfide purification system

    SciTech Connect

    1991-07-30

    A process for the purification (washing) of iron disulfide (FeS{sub 2}) powder is conducted in the Northeast corner (Area 353) of the main plant building (Building 100). This location is about 130 feet from the fenced boundary of the Partnership School/Child Development Center. In the first steps of the process, raw iron disulfide powder is ground and separated by particle size. The ground and sized powder is then purified in a three-step acid washing process using both hydrochloric acid (HCI) and hydrofluoric (HF) acid. The iron disulfide process is an intermittent batch process conducted four to eight times a year. This study is a Preliminary Hazards Assessment (PHA) to assess the hazards associated with the iron disulfide process. This is a preliminary study and will be used to determine if additional safety analysis is necessary. The scope of the PHA includes assessment of the process steps of grinding, size classification, and purification. The purpose is to identify major hazards and determine if the current and newly added safeguards are adequate for operation. The PHA also lists recommendations for additional safety features that should be added to reduce the risks of operation.

  5. Molybdenum disulfide (MoS2) nanoflakes as inherently electroactive labels for DNA hybridization detection

    NASA Astrophysics Data System (ADS)

    Loo, Adeline Huiling; Bonanni, Alessandra; Ambrosi, Adriano; Pumera, Martin

    2014-09-01

    The detection of specific DNA sequences plays a critical role in the areas of medical diagnostics, environmental monitoring, drug discovery and food safety. This has therefore become a strong driving force behind the ever-increasing demand for simple, cost-effective, highly sensitive and selective DNA biosensors. In this study, we report for the first time, a novel approach for the utilization of molybdenum disulfide nanoflakes, a member of the transition metal dichalcogenides family, in the detection of DNA hybridization. Herein, molybdenum disulfide nanoflakes serve as inherently electroactive labels, with the inherent oxidation peak exploited as the analytical signal. The principle of detection is based on the differential affinity of molybdenum disulfide nanoflakes towards single-stranded DNA and double-stranded DNA. The employment of transition metal dichalcogenide nanomaterials for sensing and biosensing purposes represents an upcoming research area which holds great promise. Hence, our findings are anticipated to have significant contributions towards the fabrication of future DNA biosensors.The detection of specific DNA sequences plays a critical role in the areas of medical diagnostics, environmental monitoring, drug discovery and food safety. This has therefore become a strong driving force behind the ever-increasing demand for simple, cost-effective, highly sensitive and selective DNA biosensors. In this study, we report for the first time, a novel approach for the utilization of molybdenum disulfide nanoflakes, a member of the transition metal dichalcogenides family, in the detection of DNA hybridization. Herein, molybdenum disulfide nanoflakes serve as inherently electroactive labels, with the inherent oxidation peak exploited as the analytical signal. The principle of detection is based on the differential affinity of molybdenum disulfide nanoflakes towards single-stranded DNA and double-stranded DNA. The employment of transition metal dichalcogenide

  6. Thiol-Disulfide Exchange Reactions in the Mammalian Extracellular Environment.

    PubMed

    Yi, Michael C; Khosla, Chaitan

    2016-06-01

    Disulfide bonds represent versatile posttranslational modifications whose roles encompass the structure, catalysis, and regulation of protein function. Due to the oxidizing nature of the extracellular environment, disulfide bonds found in secreted proteins were once believed to be inert. This notion has been challenged by the discovery of redox-sensitive disulfides that, once cleaved, can lead to changes in protein activity. These functional disulfides are twisted into unique configurations, leading to high strain and potential energy. In some cases, cleavage of these disulfides can lead to a gain of function in protein activity. Thus, these motifs can be referred to as switches. We describe the couples that control redox in the extracellular environment, examine several examples of proteins with switchable disulfides, and discuss the potential applications of disulfides in molecular biology. PMID:27023663

  7. The alkaline earth intercalates of molybdenum disulfide

    NASA Technical Reports Server (NTRS)

    Somoano, R. B.; Hadek, V.; Rembaum, A.; Samson, S.; Woollam, J. A.

    1975-01-01

    Molybdenum disulfide has been intercalated with calcium and strontium by means of the liquid ammonia technique. Chemical, X-ray, and superconductivity data are presented. The X-ray data reveal a lowering of crystal symmetry and increase of complexity of the structure upon intercalation with the alkaline earth metals. The Ca and Sr intercalates start to superconduct at 4 and 5.6 K, respectively, and show considerable anisotropy regarding the critical magnetic field.

  8. Alkali metal intercalates of molybdenum disulfide.

    NASA Technical Reports Server (NTRS)

    Somoano, R. B.; Hadek, V.; Rembaum, A.

    1973-01-01

    Study of some of the physicochemical properties of compounds obtained by subjecting natural molybdenite and single crystals of molybdenum disulfide grown by chemical vapor transport to intercalation with the alkali group of metals (Li, Na, K, Rb, and Cs) by means of the liquid ammonia technique. Reported data and results include: (1) the intercalation of the entire alkali metal group, (2) stoichiometries and X-ray data on all of the compounds, and (3) superconductivity data for all the intercalation compounds.

  9. Sterically hindered disulfide bridges in cystine diketopiperazine, cysteinyl-cysteine disulfide and derivatives.

    PubMed

    Ottnad, M; Hartter, P; Jung, G

    1975-06-01

    L-Cystine diketopiperazine (1), L-cysteinyl-cysteine disulfide -HCl (2), L-cysteinyl-cysteine disulfide methyl ester -HCl (3), and t-butyloxycarbonyl-L-cysteinyl-cysteine disulfide methyl ester (4) are investigated by CD, ultraviolet, 13C NMR, infrared and laser Raman spectroscopy. The temperature dependence of the 13C NMR signals of 1 reveals an exceptionally high energy barrier of deltaGNo. = 15.8 +/- 0.2 kcal/mol for the reversible change in helicity of the inherently dissymmetric disulfide bridge of 1. The P-helical diastereomer predominates in dimethyl-sulfoxide at 25 degrees C, with 80-85% of the molecules having this configuration. The Cotton effects of 1 are larger and show smaller temperature coefficients than the conformationally more mobile cystine compounds 2 and 3. After dissolving crystals of 1 in 95% ethanol there is a time-dependent decrease of the ellipticity of the negative Cotton effect at 225 nm, indicating a conformational change in going from crystal to solution. Besides 1, 2 and 3 are at present the only known examples of cystine derivatives with C-S-S-C torsional angles around 90 degrees, which do not exhibit optical activity in the long wavelength disulfide absorption, as is predicted for 1 from the Linderberg-Michl model. At 305 nm a new weak Cotton effect was discovered for 1. The solvent dependence of the CD spectra is discussed and the infrared and Raman spectra are assigned. PMID:1181260

  10. Cyclic disulfide C8 iminoporfiromycin: nucleophilic activation of a porfiromycin.

    PubMed

    Lee, Sang Hyup; Kohn, Harold

    2004-04-01

    The clinical success of mitomycin C (1) and its associated toxicities and resistance have led to efforts to prepare semisynthetic analogues (i.e., KW-2149 (3), BMS-181174 (4)) that have improved pharmacological profiles. In this study, we report the preparation and evaluation of the novel 7-N-(1'-amino-4',5'-dithian-2'-yl)porfiromycin C(8) cyclized imine (6) and its reference compound, 7-N-(1'-aminocyclohex-2'-yl)porfiromycin C(8) cyclized imine (13). Porfiromycin 6 contains a disulfide unit that, upon cleavage, may provide thiol(s) that affect drug reactivity. We demonstrated that phosphines dramatically accelerated 6 activation and solvolysis in methanolic solutions ("pH 7.4") compared with 13. Porfiromycins 6 and 13 efficiently cross-linked EcoRI-linearized pBR322 DNA upon addition of Et3P. We found enhanced levels of interstrand cross-link (ISC) adducts for 6 and 13 compared with porfiromycin (7) and that 6 was more efficient than 13. The large Et3P-mediated rate enhancements for the solvolysis of 6 compared with 13 and a N(7)-substituted analogue of 1, and the increased levels of ISC adducts for 6 compared with 13 and 7 are attributed to a nucleophile-assisted disulfide cleavage process that permits porfiromycin activation and nucleophile (MeOH, DNA) adduction. The in vitro antiproliferative activities of 6 and 13 using the A549 tumor cell line (lung adenocarcinoma) were determined under aerobic and hypoxic conditions and then compared with 7. Both 6 and 13 were more cytotoxic than 7, with 13 being more potent than 6. The C(8) iminoporfiromycins 6 and 13 displayed anticancer profiles similar to 3. PMID:15053618

  11. Edge-terminated molybdenum disulfide with a 9.4-Å interlayer spacing for electrochemical hydrogen production

    DOE PAGESBeta

    Gao, Min -Rui; Chan, Maria K. Y.; Sun, Yugang

    2015-07-03

    In this study, layered molybdenum disulfide has demonstrated great promise as a low-cost alternative to platinum-based catalysts for electrochemical hydrogen production from water. Research effort on this material has focused mainly on synthesizing highly nanostructured molybdenum disulfide that allows the exposure of a large fraction of active edge sites. Here we report a promising microwave-assisted strategy for the synthesis of narrow molybdenum disulfide nanosheets with edge-terminated structure and a significantly expanded interlayer spacing, which exhibit striking kinetic metrics with onset potential of -103 mV, Tafel slope of 49 mV per decade and exchange current density of 9.62 × 10-3 mAmore » cm-2, performing among the best of current molybdenum disulfide catalysts. Besides benefits from the edge-terminated structure, the expanded interlayer distance with modified electronic structure is also responsible for the observed catalytic improvement, which suggests a potential way to design newly advanced molybdenum disulfide catalysts through modulating the interlayer distance.« less

  12. Edge-terminated molybdenum disulfide with a 9.4-Å interlayer spacing for electrochemical hydrogen production

    PubMed Central

    Gao, Min-Rui; Chan, Maria K.Y.; Sun, Yugang

    2015-01-01

    Layered molybdenum disulfide has demonstrated great promise as a low-cost alternative to platinum-based catalysts for electrochemical hydrogen production from water. Research effort on this material has focused mainly on synthesizing highly nanostructured molybdenum disulfide that allows the exposure of a large fraction of active edge sites. Here we report a promising microwave-assisted strategy for the synthesis of narrow molybdenum disulfide nanosheets with edge-terminated structure and a significantly expanded interlayer spacing, which exhibit striking kinetic metrics with onset potential of −103 mV, Tafel slope of 49 mV per decade and exchange current density of 9.62 × 10−3 mA cm−2, performing among the best of current molybdenum disulfide catalysts. Besides benefits from the edge-terminated structure, the expanded interlayer distance with modified electronic structure is also responsible for the observed catalytic improvement, which suggests a potential way to design newly advanced molybdenum disulfide catalysts through modulating the interlayer distance. PMID:26138031

  13. Edge-terminated molybdenum disulfide with a 9.4-Å interlayer spacing for electrochemical hydrogen production

    SciTech Connect

    Gao, Min -Rui; Chan, Maria K. Y.; Sun, Yugang

    2015-07-03

    In this study, layered molybdenum disulfide has demonstrated great promise as a low-cost alternative to platinum-based catalysts for electrochemical hydrogen production from water. Research effort on this material has focused mainly on synthesizing highly nanostructured molybdenum disulfide that allows the exposure of a large fraction of active edge sites. Here we report a promising microwave-assisted strategy for the synthesis of narrow molybdenum disulfide nanosheets with edge-terminated structure and a significantly expanded interlayer spacing, which exhibit striking kinetic metrics with onset potential of -103 mV, Tafel slope of 49 mV per decade and exchange current density of 9.62 × 10-3 mA cm-2, performing among the best of current molybdenum disulfide catalysts. Besides benefits from the edge-terminated structure, the expanded interlayer distance with modified electronic structure is also responsible for the observed catalytic improvement, which suggests a potential way to design newly advanced molybdenum disulfide catalysts through modulating the interlayer distance.

  14. Human Defensin 5 Disulfide Array Mutants: Disulfide Bond Deletion Attenuates Antibacterial Activity Against Staphylococcus aureus

    PubMed Central

    Wanniarachchi, Yoshitha A.; Kaczmarek, Piotr; Wan, Andrea; Nolan, Elizabeth M.

    2011-01-01

    Human α-defensin 5 (HD5, HD5ox to specify the oxidized and disulfide linked form) is a 32-residue cysteine-rich host-defense peptide, expressed and released by small intestinal Paneth cells, that exhibits antibacterial activity against a number of Gram-negative and –positive bacterial strains. To ascertain the contributions of its disulfide array to structure, antimicrobial activity, and proteolytic stability, a series of HD5 double mutant peptides where pairs of cysteine residues corresponding to native disulfide linkages (Cys3—Cys31, Cys5—Cys20, Cys10—Cys30) were mutated to Ser or Ala residues were overexpressed in E. coli, purified and characterized. A hexa mutant peptide, HD5[Serhexa], where all six native Cys residues are replaced by Ser residues was also evaluated. Removal of a single native S—S linkage influences oxidative folding and regioisomerization, antibacterial activity, Gram-negative bacterial membrane permeabilization, and proteolytic stability. Whereas the majority of the HD5 mutant peptides show low-micromolar activity against Gram-negative E. coli ATCC 25922 in colony counting assays, the wild-type disulfide array is essential for low-micromolar activity against Gram-positive S. aureus ATCC 25923. Removal of a single disulfide bond attenuates the activity observed for HD5ox against this Gram-positive bacterial strain. This observation supports the notion that the HD5ox mechanism of antibacterial action differs for Gram-negative and Gram-positive species (Wei, G.; de Leeuw, E., Pazgier, M., Yuan, W., Zou, G., Wang, J., Ericksen, B., Lu, W.-Y.; Lehrer, R. I.; Lu, W. (2009) J. Biol. Chem. 284, 29180-29192), and that the native disulfide array is a requirement for its activity against S. aureus. PMID:21861459

  15. Engineering de novo disulfide bond in bacterial α-type carbonic anhydrase for thermostable carbon sequestration

    NASA Astrophysics Data System (ADS)

    Jo, Byung Hoon; Park, Tae Yoon; Park, Hyun June; Yeon, Young Joo; Yoo, Young Je; Cha, Hyung Joon

    2016-07-01

    Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial α-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 °C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions.

  16. Engineering de novo disulfide bond in bacterial α-type carbonic anhydrase for thermostable carbon sequestration

    PubMed Central

    Jo, Byung Hoon; Park, Tae Yoon; Park, Hyun June; Yeon, Young Joo; Yoo, Young Je; Cha, Hyung Joon

    2016-01-01

    Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial α-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 °C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions. PMID:27385052

  17. Disulfide bonds and glycosylation in fungal peroxidases.

    PubMed

    Limongi, P; Kjalke, M; Vind, J; Tams, J W; Johansson, T; Welinder, K G

    1995-01-15

    Four conserved disulfide bonds and N-linked and O-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from Trametes versicolor, and from Coprinus cinereus peroxidase (CIP) and recombinant C. cinereus peroxidase (rCIP) expressed in Aspergillus oryzae. The eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. CIP and rCIP were identical in their glycosylation pattern, although the extent of glycan chain heterogeneity depended on the fermentation batch. CIP and rCIP have one N-linked glycan composed only of GlcNAc and Man at residue Asn142, and two O-linked glycans near the C-terminus. The major glycoform consists of single Man residues at Thr331 and at Ser338. T. versicolor lignin isoperoxidase TvLP10 contains a single N-linked glycan composed of (GlcNAc)2Man5 bound to Asn103, whereas (GlcNAc)2Man3 was found in T. versicolor manganese isoperoxidase TvMP2 at the same position. In addition, mass spectrometry of the C-terminal peptide of TvMP2 indicated the presence of five Man residues in O-linked glycans. No phosphate was found in these fungal peroxidases. PMID:7851395

  18. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  19. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  20. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  1. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  2. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  3. Converting a Sulfenic Acid Reductase into a Disulfide Bond Isomerase

    PubMed Central

    Chatelle, Claire; Kraemer, Stéphanie; Ren, Guoping; Chmura, Hannah; Marechal, Nils; Boyd, Dana; Roggemans, Caroline; Ke, Na; Riggs, Paul; Bardwell, James

    2015-01-01

    Abstract Aims: Posttranslational formation of disulfide bonds is essential for the folding of many secreted proteins. Formation of disulfide bonds in a protein with more than two cysteines is inherently fraught with error and can result in incorrect disulfide bond pairing and, consequently, misfolded protein. Protein disulfide bond isomerases, such as DsbC of Escherichia coli, can recognize mis-oxidized proteins and shuffle the disulfide bonds of the substrate protein into their native folded state. Results: We have developed a simple blue/white screen that can detect disulfide bond isomerization in vivo, using a mutant alkaline phosphatase (PhoA*) in E. coli. We utilized this screen to isolate mutants of the sulfenic acid reductase (DsbG) that allowed this protein to act as a disulfide bond isomerase. Characterization of the isolated mutants in vivo and in vitro allowed us to identify key amino acid residues responsible for oxidoreductase properties of thioredoxin-like proteins such as DsbC or DsbG. Innovation and Conclusions: Using these key residues, we also identified and characterized interesting environmental homologs of DsbG with novel properties, thus demonstrating the capacity of this screen to discover and elucidate mechanistic details of in vivo disulfide bond isomerization. Antioxid. Redox Signal. 23, 945–957. PMID:26191605

  4. Early events in the disulfide-coupled folding of BPTI.

    PubMed Central

    Bulaj, G.; Goldenberg, D. P.

    1999-01-01

    Recent studies of the refolding of reduced bovine pancreatic trypsin inhibitor (BPTI) have shown that a previously unidentified intermediate with a single disulfide is formed much more rapidly than any other one-disulfide species. This intermediate contains a disulfide that is present in the native protein (between Cys14 and 38), but it is thermodynamically less stable than the other two intermediates with single native disulfides. To characterize the role of the [14-38] intermediate and the factors that favor its formation, detailed kinetic and mutational analyses of the early disulfide-formation steps were carried out. The results of these studies indicate that the formation of [14-38] from the fully reduced protein is favored by both local electrostatic effects, which enhance the reactivities of the Cys14 and 38 thiols, and conformational tendencies that are diminished by the addition of urea and are enhanced at lower temperatures. At 25 degrees C and pH 7.3, approximately 35% of the reduced molecules were found to initially form the 14-38 disulfide, but the majority of these molecules then undergo intramolecular rearrangements to generate non-native disulfides, and subsequently the more stable intermediates with native disulfides. Amino acid replacements, other than those involving Cys residues, were generally found to have only small effects on either the rate of forming [14-38] or its thermodynamic stability, even though many of the same substitutions greatly destabilized the native protein and other disulfide-bonded intermediates. In addition, those replacements that did decrease the steady-state concentration of [14-38] did not adversely affect further folding and disulfide formation. These results suggest that the weak and transient interactions that are often detected in unfolded proteins and early folding intermediates may, in some cases, not persist or promote subsequent folding steps. PMID:10493584

  5. [Child sexual exploitation].

    PubMed

    Cabello, María F; Castaldi, Paula D; Cataldo, Andrea M

    2009-01-01

    Child Sexual Exploitation is a complex phenomenon in our country and the world; it dates back to an ancient past but it has a very recent conceptualization and specific approach. This article proposes a tour through this process as well as some inputs for its categorization, the attention to the affected subjects by the very design of public policies taken from a concrete institutional experience. PMID:19812796

  6. Sequential bottom-up and top-down processing for the synthesis of transition metal dichalcogenide nanosheets: the case of rhenium disulfide (ReS2).

    PubMed

    Al-Dulaimi, Naktal; Lewis, Edward A; Lewis, David J; Howell, Simon K; Haigh, Sarah J; O'Brien, Paul

    2016-06-14

    Bottom-up (aerosol-assisted chemical vapor deposition, AACVD) and top-down (liquid phase exfoliation, LPE) processing methodologies are used in tandem to produce colloids of few-layer thick rhenium disulfide (ReS2) in N-methyl pyrrolidone. The processing route we use is a potentially robust and scalable pathway to manufacture useful 2D materials. PMID:27250595

  7. Advances in rechargeable lithium molybdenum disulfide batteries

    NASA Technical Reports Server (NTRS)

    Brandt, K.; Stiles, J. A. R.

    1985-01-01

    The lithium molybdenum disulfide system as demonstrated in a C size cell, offers performance characteristics for applications where light weight and low volume are important. A gravimetric energy density of 90 watt hours per kilogram can be achieved in a C size cell package. The combination of charge retention capabilities, high energy density and a state of charge indicator in a rechargeable cell provides power package for a wide range of devices. The system overcomes the memory effect in Nicads where the full capacity of the battery cannot be utilized unless it was utilized on previous cycles. The development of cells with an advanced electrolyte formulation led to an improved rate capability especially at low temperatures and to a significantly improved life cycle.

  8. Superconductivity in highly disordered dense carbon disulfide

    PubMed Central

    Dias, Ranga P.; Yoo, Choong-Shik; Struzhkin, Viktor V.; Kim, Minseob; Muramatsu, Takaki; Matsuoka, Takahiro; Ohishi, Yasuo; Sinogeikin, Stanislav

    2013-01-01

    High pressure plays an increasingly important role in both understanding superconductivity and the development of new superconducting materials. New superconductors were found in metallic and metal oxide systems at high pressure. However, because of the filled close-shell configuration, the superconductivity in molecular systems has been limited to charge-transferred salts and metal-doped carbon species with relatively low superconducting transition temperatures. Here, we report the low-temperature superconducting phase observed in diamagnetic carbon disulfide under high pressure. The superconductivity arises from a highly disordered extended state (CS4 phase or phase III[CS4]) at ∼6.2 K over a broad pressure range from 50 to 172 GPa. Based on the X-ray scattering data, we suggest that the local structural change from a tetrahedral to an octahedral configuration is responsible for the observed superconductivity. PMID:23818624

  9. Pressure-induced metallization of molybdenum disulfide.

    PubMed

    Chi, Zhen-Hua; Zhao, Xiao-Miao; Zhang, Haidong; Goncharov, Alexander F; Lobanov, Sergey S; Kagayama, Tomoko; Sakata, Masafumi; Chen, Xiao-Jia

    2014-07-18

    X-ray diffraction, Raman spectroscopy, and electrical conductivity measurements of molybdenum disulfide MoS(2) are performed at pressures up to 81 GPa in diamond anvil cells. Above 20 GPa, we find discontinuous changes in Raman spectra and x-ray diffraction patterns which provide evidence for isostructural phase transition from 2H(c) to 2H(a) modification through layer sliding previously predicted theoretically. This first-order transition, which is completed around 40 GPa, is characterized by a collapse in the c-lattice parameter and volume and also by changes in interlayer bonding. After the phase transition completion, MoS(2) becomes metallic. The reversibility of the phase transition is identified from all these techniques. PMID:25083660

  10. Circulation of oligonucleotides by disulfide bridge formation.

    PubMed Central

    Gao, H; Yang, M; Patel, R; Cook, A F

    1995-01-01

    An effective, convenient method for the circularization of oligonucleotides has been developed. This procedure involved preparation of an oligonucleotide with backbone-linked 5'- and 3'-terminal hexamethylenethiol groups, followed by oxidation of the thiol groups with air of oxygen to produce the corresponding circular sequence bridged via a bis(hexamethylene)-disulfide moiety. The method has been applied to the circularization of oligodeoxynucleotide sequences of varying lengths (5, 10, 15, 20, 30 and 40 bases), and the circularization process was highly efficient as shown by HPLC or gel electrophoresis of the crude reaction mixtures. Competing reactions such as dimerization were not significant except for the longer sequences (30 and 40 bases). The circularization of an eight base RNA sequence was also accomplished, as well as hexa-ethylene glycol bridged poly-T sequences capable of triplex formation. PMID:7596832

  11. Ultrafast response of monolayer molybdenum disulfide photodetectors

    PubMed Central

    Wang, Haining; Zhang, Changjian; Chan, Weimin; Tiwari, Sandip; Rana, Farhan

    2015-01-01

    The strong light emission and absorption exhibited by single atomic layer transitional metal dichalcogenides in the visible to near-infrared wavelength range make them attractive for optoelectronic applications. In this work, using two-pulse photovoltage correlation technique, we show that monolayer molybdenum disulfide photodetector can have intrinsic response times as short as 3 ps implying photodetection bandwidths as wide as 300 GHz. The fast photodetector response is a result of the short electron–hole and exciton lifetimes in this material. Recombination of photoexcited carriers in most two-dimensional metal dichalcogenides is dominated by nonradiative processes, most notable among which is Auger scattering. The fast response time, and the ease of fabrication of these devices, make them interesting for low-cost ultrafast optical communication links. PMID:26572726

  12. Ferromagnetism in exfoliated tungsten disulfide nanosheets

    PubMed Central

    2013-01-01

    Two-dimensional-layered transition metal dichalcogenides nanosheets have attracted tremendous attention for their promising applications in spintronics because the atomic-thick nanosheets can not only enhance the intrinsic properties of their bulk counterparts, but also give birth to new promising properties. In this paper, ultrathin tungsten disulfide (WS2) nanosheets were gotten by liquid exfoliation route from its bulk form using dimethylformamide (DMF). Compared to the antiferromagnetism bulk WS2, ultrathin WS2 nanosheets show intrinsic room-temperature ferromagnetism (FM) with the maximized saturation magnetization of 0.004 emu/g at 10 K, where the appearance of FM in the nanosheets is partly due to the presence of zigzag edges in the magnetic ground state at the grain boundaries. PMID:24134699

  13. Enzyme structure captures four cysteines aligned for disulfide relay

    PubMed Central

    Gat, Yair; Vardi-Kilshtain, Alexandra; Grossman, Iris; Major, Dan Thomas; Fass, Deborah

    2014-01-01

    Thioredoxin superfamily proteins introduce disulfide bonds into substrates, catalyze the removal of disulfides, and operate in electron relays. These functions rely on one or more dithiol/disulfide exchange reactions. The flavoenzyme quiescin sulfhydryl oxidase (QSOX), a catalyst of disulfide bond formation with an interdomain electron transfer step in its catalytic cycle, provides a unique opportunity for exploring the structural environment of enzymatic dithiol/disulfide exchange. Wild-type Rattus norvegicus QSOX1 (RnQSOX1) was crystallized in a conformation that juxtaposes the two redox-active di-cysteine motifs in the enzyme, presenting the entire electron-transfer pathway and proton-transfer participants in their native configurations. As such a state cannot generally be enriched and stabilized for analysis, RnQSOX1 gives unprecedented insight into the functional group environments of the four cysteines involved in dithiol/disulfide exchange and provides the framework for analysis of the energetics of electron transfer in the presence of the bound flavin adenine dinucleotide cofactor. Hybrid quantum mechanics/molecular mechanics (QM/MM) free energy simulations based on the X-ray crystal structure suggest that formation of the interdomain disulfide intermediate is highly favorable and secures the flexible enzyme in a state from which further electron transfer via the flavin can occur. PMID:24888638

  14. Multisensor staring exploitation

    NASA Astrophysics Data System (ADS)

    Bryant, Michael L.

    2008-04-01

    The focus of this paper is on the exploitation of new staring sensors to address the urban surveillance challenge and help combat the war on terror. A staring sensor visualization environment, known as the Data Table, will be presented which integrates staring sensors with close-in sensors, such as small UAVs, building mounted sensors, and unattended ground sensors (UGS). There are several staring sensors in development, but two in particular will be highlighted in this paper - NightStare and the Gotcha Radar, both under development by the Air Force Research Laboratory (AFRL).

  15. Disulfide Bond Oxidoreductase DsbA2 of Legionella pneumophila Exhibits Protein Disulfide Isomerase Activity

    PubMed Central

    Kpadeh, Zegbeh Z.; Jameson-Lee, Max; Yeh, Anthony J.; Chertihin, Olga; Shumilin, Igor A.; Dey, Rafik; Day, Shandra R.

    2013-01-01

    The extracytoplasmic assembly of the Dot/Icm type IVb secretion system (T4SS) of Legionella pneumophila is dependent on correct disulfide bond (DSB) formation catalyzed by a novel and essential disulfide bond oxidoreductase DsbA2 and not by DsbA1, a second nonessential DSB oxidoreductase. DsbA2, which is widely distributed in the microbial world, is phylogenetically distinct from the canonical DsbA oxidase and the DsbC protein disulfide isomerase (PDI)/reductase of Escherichia coli. Here we show that the extended N-terminal amino acid sequence of DsbA2 (relative to DsbA proteins) contains a highly conserved 27-amino-acid dimerization domain enabling the protein to form a homodimer. Complementation tests with E. coli mutants established that L. pneumophila dsbA1, but not the dsbA2 strain, restored motility to a dsbA mutant. In a protein-folding PDI detector assay, the dsbA2 strain, but not the dsbA1 strain, complemented a dsbC mutant of E. coli. Deletion of the dimerization domain sequences from DsbA2 produced the monomer (DsbA2N), which no longer exhibited PDI activity but complemented the E. coli dsbA mutant. PDI activity was demonstrated in vitro for DsbA2 but not DsbA1 in a nitrocefin-based mutant TEM β-lactamase folding assay. In an insulin reduction assay, DsbA2N activity was intermediate between those of DsbA2 and DsbA1. In L. pneumophila, DsbA2 was maintained as a mixture of thiol and disulfide forms, while in E. coli, DsbA2 was present as the reduced thiol. Our studies suggest that DsbA2 is a naturally occurring bifunctional disulfide bond oxidoreductase that may be uniquely suited to the majority of intracellular bacterial pathogens expressing T4SSs as well as in many slow-growing soil and aquatic bacteria. PMID:23435972

  16. Radical cations of sulfides and disulfides: An ESR study

    SciTech Connect

    Bonazzola, L.; Michaut, J.P.; Roncin, J.

    1985-09-15

    Exposure of dilute solutions of dimethylsulfide, methanethiol, tetrahydrothiophene, terbutyl and diterbutyl-sulfides, dimethyl-disulfide, and diterbutyldisulfide, in freon at 77 K to /sup 60/Co ..gamma.. rays gave the corresponding cations. From the reported ESR spectra, g tensors were obtained. It was found that both sulfide and disulfide cations exhibit the same g tensor: (g/sub max/ = 2.034 +- 0.002, g/sub int/ = 2.017 +- 0.001, g/sub min/ = 2.001 +- 0.005). From this result it has been shown that the disulfide cation is planar. This finding was supported by fully optimized geometry ab initio calculations.

  17. Protein disulfide isomerase a multifunctional protein with multiple physiological roles

    NASA Astrophysics Data System (ADS)

    Ali Khan, Hyder; Mutus, Bulent

    2014-08-01

    Protein disulfide isomerase (PDI), is a member of the thioredoxin superfamily of redox proteins. PDI has three catalytic activities including, thiol-disulfide oxireductase, disulfide isomerase and redox-dependent chaperone. Originally, PDI was identified in the lumen of the endoplasmic reticulum and subsequently detected at additional locations, such as cell surfaces and the cytosol. This review will provide an overview of the recent advances in relating the structural features of PDI to its multiple catalytic roles as well as its physiological and pathophysiological functions related to redox regulation and protein folding.

  18. Transnational gestational surrogacy: does it have to be exploitative?

    PubMed

    Kirby, Jeffrey

    2014-01-01

    This article explores the controversial practice of transnational gestational surrogacy and poses a provocative question: Does it have to be exploitative? Various existing models of exploitation are considered and a novel exploitation-evaluation heuristic is introduced to assist in the analysis of the potentially exploitative dimensions/elements of complex health-related practices. On the basis of application of the heuristic, I conclude that transnational gestational surrogacy, as currently practiced in low-income country settings (such as rural, western India), is exploitative of surrogate women. Arising out of consideration of the heuristic's exploitation conditions, a set of public education and enabled choice, enhanced protections, and empowerment reforms to transnational gestational surrogacy practice is proposed that, if incorporated into a national regulatory framework and actualized within a low income country, could possibly render such practice nonexploitative. PMID:24766117

  19. 21 CFR 520.1802a - Piperazine-carbon disulfide complex suspension.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Piperazine-carbon disulfide complex suspension... § 520.1802a Piperazine-carbon disulfide complex suspension. (a) Specifications. Each fluid ounce of suspension contains 7.5 grams of piperazine-carbon disulfide complex. The piperazine-carbon disulfide...

  20. 21 CFR 520.1802a - Piperazine-carbon disulfide complex suspension.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Piperazine-carbon disulfide complex suspension... § 520.1802a Piperazine-carbon disulfide complex suspension. (a) Specifications. Each fluid ounce of suspension contains 7.5 grams of piperazine-carbon disulfide complex. The piperazine-carbon disulfide...

  1. 21 CFR 520.1802a - Piperazine-carbon disulfide complex suspension.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Piperazine-carbon disulfide complex suspension... § 520.1802a Piperazine-carbon disulfide complex suspension. (a) Specifications. Each fluid ounce of suspension contains 7.5 grams of piperazine-carbon disulfide complex. The piperazine-carbon disulfide...

  2. 21 CFR 520.1802a - Piperazine-carbon disulfide complex suspension.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Piperazine-carbon disulfide complex suspension... § 520.1802a Piperazine-carbon disulfide complex suspension. (a) Specifications. Each fluid ounce of suspension contains 7.5 grams of piperazine-carbon disulfide complex. The piperazine-carbon disulfide...

  3. 21 CFR 520.1802a - Piperazine-carbon disulfide complex suspension.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Piperazine-carbon disulfide complex suspension... § 520.1802a Piperazine-carbon disulfide complex suspension. (a) Specifications. Each fluid ounce of suspension contains 7.5 grams of piperazine-carbon disulfide complex. The piperazine-carbon disulfide...

  4. Moving target exploitation

    NASA Astrophysics Data System (ADS)

    Johnson, Bruce L.; Grayson, Timothy P.

    1998-08-01

    The understanding of maneuvering forces is invaluable to the warfighter, as it enhances understanding of enemy force structure and disposition, provides cues to potential enemy actions, and expedites targeting of time critical targets. Airborne ground moving target indicator (GMTI) radars are a class of highly-effective, all-weather, wide-area senors that aid in the surveillance of these moving ground vehicles. Unfortunately conventional GMTI radars are incapable of identifying individual vehicles, and techniques for exploiting information imbedded within GMTI radar reports are limited. The Defense Advanced Research Projects Agency (DARPA) Moving Target Exploitation (MTE) program is working to mitigate these deficiencies by developing, integrating, and evaluating a suite of automated and semi-automated technologies to classify moving targets and units, and to provide indications of their activities. These techniques include: aid in the interpretation of GMTI data to provide moving force structure analysis, automatic tracking of thousands of moving ground vehicles, 1-D target classification based upon high-range- resolution (HRR) radar profiles, and 2-D target classification based upon moving target imaging (MTIm) synthetic aperture radar (SAR). This paper shall present the MTE concept and motivation and provide an overview of results to date.

  5. Multilevel fusion exploitation

    NASA Astrophysics Data System (ADS)

    Lindberg, Perry C.; Dasarathy, Belur V.; McCullough, Claire L.

    1996-06-01

    This paper describes a project that was sponsored by the U.S. Army Space and Strategic Defense Command (USASSDC) to develop, test, and demonstrate sensor fusion algorithms for target recognition. The purpose of the project was to exploit the use of sensor fusion at all levels (signal, feature, and decision levels) and all combinations to improve target recognition capability against tactical ballistic missile (TBM) targets. These algorithms were trained with simulated radar signatures to accurately recognize selected TBM targets. The simulated signatures represent measurements made by two radars (S-band and X- band) with the targets at a variety of aspect and roll angles. Two tests were conducted: one with simulated signatures collected at angles different from those in the training database and one using actual test data. The test results demonstrate a high degree of recognition accuracy. This paper describes the training and testing techniques used; shows the fusion strategy employed; and illustrates the advantages of exploiting multi-level fusion.

  6. Improved molybdenum disulfide-silver motor brushes have extended life

    NASA Technical Reports Server (NTRS)

    Horton, J. C.; King, H. M.

    1964-01-01

    Motor brushes of proper quantities of molybdenum disulfide and copper or silver are manufactured by sintering techniques. Graphite molds are used. These brushes operate satisfactorily for long periods in normal atmosphere or in a high-vacuum environment.

  7. Biotechnological exploitation of microalgae.

    PubMed

    Gangl, Doris; Zedler, Julie A Z; Rajakumar, Priscilla D; Martinez, Erick M Ramos; Riseley, Anthony; Włodarczyk, Artur; Purton, Saul; Sakuragi, Yumiko; Howe, Christopher J; Jensen, Poul Erik; Robinson, Colin

    2015-12-01

    Microalgae are a diverse group of single-cell photosynthetic organisms that include cyanobacteria and a wide range of eukaryotic algae. A number of microalgae contain high-value compounds such as oils, colorants, and polysaccharides, which are used by the food additive, oil, and cosmetic industries, among others. They offer the potential for rapid growth under photoautotrophic conditions, and they can grow in a wide range of habitats. More recently, the development of genetic tools means that a number of species can be transformed and hence used as cell factories for the production of high-value chemicals or recombinant proteins. In this article, we review exploitation use of microalgae with a special emphasis on genetic engineering approaches to develop cell factories, and the use of synthetic ecology approaches to maximize productivity. We discuss the success stories in these areas, the hurdles that need to be overcome, and the potential for expanding the industry in general. PMID:26400987

  8. The Geohazards Exploitation Platform

    NASA Astrophysics Data System (ADS)

    Laur, Henri; Casu, Francesco; Bally, Philippe; Caumont, Hervé; Pinto, Salvatore

    2016-04-01

    The Geohazards Exploitation Platform, or Geohazards TEP (GEP), is an ESA originated R&D activity of the EO ground segment to demonstrate the benefit of new technologies for large scale processing of EO data. This encompasses on-demand processing for specific user needs, systematic processing to address common information needs of the geohazards community, and integration of newly developed processors for scientists and other expert users. The platform supports the geohazards community's objectives as defined in the context of the International Forum on Satellite EO and Geohazards organised by ESA and GEO in Santorini in 2012. The GEP is a follow on to the Supersites Exploitation Platform (SSEP) an ESA initiative to support the Geohazards Supersites & Natural Laboratories initiative (GSNL). Today the GEP allows to exploit 70+ Terabyte of ERS and ENVISAT archive and the Copernicus Sentinel-1 data available on line. The platform has already engaged 22 European early adopters in a validation activity initiated in March 2015. Since September, this validation has reached 29 single user projects. Each project is concerned with either integrating an application, running on demand processing or systematically generating a product collection using an application available in the platform. The users primarily include 15 geoscience centres and universities based in Europe: British Geological Survey (UK), University of Leeds (UK), University College London (UK), ETH University of Zurich (CH), INGV (IT), CNR-IREA and CNR-IRPI (IT), University of L'Aquila (IT), NOA (GR), Univ. Blaise Pascal & CNRS (FR), Ecole Normale Supérieure (FR), ISTERRE / University of Grenoble-Alpes (FR). In addition, there are users from Africa and North America with the University of Rabat (MA) and the University of Miami (US). Furthermore two space agencies and four private companies are involved: the German Space Research Centre DLR (DE), the European Space Agency (ESA), Altamira Information (ES

  9. Exploiting Endocytosis for Nanomedicines

    PubMed Central

    Akinc, Akin; Battaglia, Giuseppe

    2013-01-01

    In this article, we briefly review the endocytic pathways used by cells, pointing out their defining characteristics and highlighting physical limitations that may direct the internalization of nanoparticles to a subset of these pathways. A more detailed description of these pathways is presented in the literature. We then focus on the endocytosis of nanomedicines and present how various nanomaterial parameters impact these endocytic processes. This topic is an area of active research, motivated by the recognition that an improved understanding of how nanomaterials interact at the molecular, cellular, and whole-organism level will lead to the design of better nanomedicines in the future. Next, we briefly review some of the important nanomedicines already on the market or in clinical development that serve to exemplify how endocytosis can be exploited for medical benefit. Finally, we present some key unanswered questions and remaining challenges to be addressed by the field. PMID:24186069

  10. Image exploitation for MISAR

    NASA Astrophysics Data System (ADS)

    Heinze, N.; Edrich, M.; Saur, G.; Krüger, W.

    2007-04-01

    The miniature SAR-system MiSAR has been developed by EADS Germany for lightweight UAVs like the LUNASystem. MiSAR adds to these tactical UAV-systems the all-weather reconnaissance capability, which is missing until now. Unlike other SAR sensors, that produce large strip maps at update rates of several seconds, MiSAR generates sequences of SAR images with approximately 1 Hz frame rate. photo interpreters (PI) of tactical drones, now mainly experienced with visual interpretation, are not used to SARimages, especially not with SAR-image sequence characteristics. So they should be supported to improve their ability to carry out their task with a new, demanding sensor system. We have therefore analyzed and discussed with military PIs in which task MiSAR can be used and how the PIs can be supported by special algorithms. We developed image processing- and exploitation-algorithms for such SAR-image sequences. A main component is the generation of image sequence mosaics to get more oversight. This mosaicing has the advantage that also non straight /linear flight-paths and varying squint angles can be processed. Another component is a screening-component for manmade objects to mark regions of interest in the image sequences. We use a classification based approach, which can be easily adapted to new sensors and scenes. These algorithms are integrated into an image exploitation system to improve the image interpreters ability to get a better oversight, better orientation and helping them to detect relevant objects, especially considering long endurance reconnaissance missions.

  11. Disulfide Mispairing During Proinsulin Folding in the Endoplasmic Reticulum.

    PubMed

    Haataja, Leena; Manickam, Nandini; Soliman, Ann; Tsai, Billy; Liu, Ming; Arvan, Peter

    2016-04-01

    Proinsulin folding within the endoplasmic reticulum (ER) remains incompletely understood, but it is clear that in mutant INS gene-induced diabetes of youth (MIDY), progression of the (three) native disulfide bonds of proinsulin becomes derailed, causing insulin deficiency, β-cell ER stress, and onset of diabetes. Herein, we have undertaken a molecular dissection of proinsulin disulfide bond formation, using bioengineered proinsulins that can form only two (or even only one) of the native proinsulin disulfide bonds. In the absence of preexisting proinsulin disulfide pairing, Cys(B19)-Cys(A20) (a major determinant of ER stress response activation and proinsulin stability) preferentially initiates B-A chain disulfide bond formation, whereas Cys(B7)-Cys(A7) can initiate only under oxidizing conditions beyond that existing within the ER of β-cells. Interestingly, formation of these two "interchain" disulfide bonds demonstrates cooperativity, and together, they are sufficient to confer intracellular transport competence to proinsulin. The three most common proinsulin disulfide mispairings in the ER appear to involve Cys(A11)-Cys(A20), Cys(A7)-Cys(A20), and Cys(B19)-Cys(A11), each disrupting the critical Cys(B19)-Cys(A20) pairing. MIDY mutations inhibit Cys(B19)-Cys(A20) formation, but treatment to force oxidation of this disulfide bond improves folding and results in a small but detectable increase of proinsulin export. These data suggest possible therapeutic avenues to ameliorate ER stress and diabetes. PMID:26822090

  12. Thiosaccharine disulfide: Synthesis, crystal structure, spectroscopic characterization and theoretical study

    NASA Astrophysics Data System (ADS)

    Ferullo, Ricardo M.; Granados, Alejandro; Lanterna, Anabel; Güida, Jorge A.; Piro, Oscar E.; Castellano, Eduardo E.; Dennehy, Mariana

    2013-01-01

    The title compound, (thiosaccharine disulfide), bis[1,1'dioxide-2,3-dihidro-1,2-benzoisothiazol]disulfide, (tsac)2 has been synthesized and fully characterized by UV-Visible, IR, Raman, 1H and 13C NMR spectroscopy elemental analysis and structural X-ray crystallography. A DFT theoretical study has been performed and good agreement between experimental and theoretical values of structural parameters and vibration frequencies have been achieved.

  13. Disulfide-Bond-Forming Pathways in Gram-Positive Bacteria

    PubMed Central

    2015-01-01

    Disulfide bonds are important for the stability and function of many secreted proteins. In Gram-negative bacteria, these linkages are catalyzed by thiol-disulfide oxidoreductases (Dsb) in the periplasm. Protein oxidation has been well studied in these organisms, but it has not fully been explored in Gram-positive bacteria, which lack traditional periplasmic compartments. Recent bioinformatics analyses have suggested that the high-GC-content bacteria (i.e., actinobacteria) rely on disulfide-bond-forming pathways. In support of this, Dsb-like proteins have been identified in Mycobacterium tuberculosis, but their functions are not known. Actinomyces oris and Corynebacterium diphtheriae have recently emerged as models to study disulfide bond formation in actinobacteria. In both organisms, disulfide bonds are catalyzed by the membrane-bound oxidoreductase MdbA. Remarkably, unlike known Dsb proteins, MdbA is important for pathogenesis and growth, which makes it a potential target for new antibacterial drugs. This review will discuss disulfide-bond-forming pathways in bacteria, with a special focus on Gram-positive bacteria. PMID:26644434

  14. The Unfolded Protein Response and the Role of Protein Disulfide Isomerase in Neurodegeneration

    PubMed Central

    Perri, Emma R.; Thomas, Colleen J.; Parakh, Sonam; Spencer, Damian M.; Atkin, Julie D.

    2016-01-01

    The maintenance and regulation of proteostasis is a critical function for post-mitotic neurons and its dysregulation is increasingly implicated in neurodegenerative diseases. Despite having different clinical manifestations, these disorders share similar pathology; an accumulation of misfolded proteins in neurons and subsequent disruption to cellular proteostasis. The endoplasmic reticulum (ER) is an important component of proteostasis, and when the accumulation of misfolded proteins occurs within the ER, this disturbs ER homeostasis, giving rise to ER stress. This triggers the unfolded protein response (UPR), distinct signaling pathways that whilst initially protective, are pro-apoptotic if ER stress is prolonged. ER stress is increasingly implicated in neurodegenerative diseases, and emerging evidence highlights the complexity of the UPR in these disorders, with both protective and detrimental components being described. Protein Disulfide Isomerase (PDI) is an ER chaperone induced during ER stress that is responsible for the formation of disulfide bonds in proteins. Whilst initially considered to be protective, recent studies have revealed unconventional roles for PDI in neurodegenerative diseases, distinct from its normal function in the UPR and the ER, although these mechanisms remain poorly defined. However, specific aspects of PDI function may offer the potential to be exploited therapeutically in the future. This review will focus on the evidence linking ER stress and the UPR to neurodegenerative diseases, with particular emphasis on the emerging functions ascribed to PDI in these conditions. PMID:26779479

  15. User interface development for semiautomated imagery exploitation

    NASA Astrophysics Data System (ADS)

    O'Connor, R. P.; Bohling, Edward H.

    1991-08-01

    Operational reconnaissance technical organizations are burdened by greatly increasing workloads due to expanding capabilities for collection and delivery of large-volume near-real- time multisensor/multispectral softcopy imagery. Related to the tasking of reconnaissance platforms to provide the imagery are more stringent timelines for exploiting the imagery in response to the rapidly changing threat environment being monitored. The development of a semi-automated softcopy multisensor image exploitation capability is a critical step toward integrating existing advanced image processing techniques in conjunction with appropriate intelligence and cartographic data for next-generation image exploitation systems. This paper discusses the results of a recent effort to develop computer-assisted aids for the image analyst (IA) in order to rapidly and accurately exploit multispectral/multisensor imagery in combination with intelligence support data and cartographic information for the purpose of target detection and identification. A key challenge of the effort was to design and implement an effective human-computer interface that would satisfy any generic IA task and readily accommodate the needs of a broad range of IAs.

  16. Intrinsic structural defects in monolayer molybdenum disulfide

    SciTech Connect

    Zhou, Wu; Idrobo Tapia, Juan C

    2013-01-01

    Monolayer molybdenum disulfide (MoS2) is a two-dimensional direct band gap semiconductor with distinctive mechanical, electronic, optical and chemical properties that can be utilized for novel nanoelectronics and optoelectronics devices. The performance of these electronic devices strongly depends on the quality and defect morphology of the MoS2 layers. Yet, little is known about the atomic structure of defects present in monolayer MoS2 and their influences on the material properties. Here we provide a systematic study of various intrinsic structural defects, including point defects, grain boundaries, and edges, in chemical vapor phase grown monolayer MoS2 via direct atomic resolution imaging, and explore their energy landscape and electronic properties using first-principles calculations. We discover that one-dimensional metallic wires can be created via two different types of 60 grain boundaries consisting of distinct 4-fold ring chains. A new type of edge reconstruction, representing a transition state during growth, was also identified, providing insights into the material growth mechanism. The atomic scale study of structural defects presented here brings new opportunities to tailor the properties of MoS2 via controlled synthesis and defect engineering.

  17. Scalable Production of Molybdenum Disulfide Based Biosensors.

    PubMed

    Naylor, Carl H; Kybert, Nicholas J; Schneier, Camilla; Xi, Jin; Romero, Gabriela; Saven, Jeffery G; Liu, Renyu; Johnson, A T Charlie

    2016-06-28

    We demonstrate arrays of opioid biosensors based on chemical vapor deposition grown molybdenum disulfide (MoS2) field effect transistors (FETs) coupled to a computationally redesigned, water-soluble variant of the μ-opioid receptor (MOR). By transferring dense films of monolayer MoS2 crystals onto prefabricated electrode arrays, we obtain high-quality FETs with clean surfaces that allow for reproducible protein attachment. The fabrication yield of MoS2 FETs and biosensors exceeds 95%, with an average mobility of 2.0 cm(2) V(-1) s(-1) (36 cm(2) V(-1) s(-1)) at room temperature under ambient (in vacuo). An atomic length nickel-mediated linker chemistry enables target binding events that occur very close to the MoS2 surface to maximize sensitivity. The biosensor response calibration curve for a synthetic opioid peptide known to bind to the wild-type MOR indicates binding affinity that matches values determined using traditional techniques and a limit of detection ∼3 nM (1.5 ng/mL). The combination of scalable array fabrication and rapid, precise binding readout enabled by the MoS2 transistor offers the prospect of a solid-state drug testing platform for rapid readout of the interactions between novel drugs and their intended protein targets. PMID:27227361

  18. Monoclonal antibody disulfide reduction during manufacturing

    PubMed Central

    Hutterer, Katariina M.; Hong, Robert W.; Lull, Jonathon; Zhao, Xiaoyang; Wang, Tian; Pei, Rex; Le, M. Eleanor; Borisov, Oleg; Piper, Rob; Liu, Yaoqing Diana; Petty, Krista; Apostol, Izydor; Flynn, Gregory C.

    2013-01-01

    Manufacturing-induced disulfide reduction has recently been reported for monoclonal human immunoglobulin gamma (IgG) antibodies, a widely used modality in the biopharmaceutical industry. This effect has been tied to components of the intracellular thioredoxin reduction system that are released upon cell breakage. Here, we describe the effect of process parameters and intrinsic molecule properties on the extent of reduction. Material taken from cell cultures at the end of production displayed large variations in the extent of antibody reduction between different products, including no reduction, when subjected to the same reduction-promoting harvest conditions. Additionally, in a reconstituted model in which process variables could be isolated from product properties, we found that antibody reduction was dependent on the cell line (clone) and cell culture process. A bench-scale model using a thioredoxin/thioredoxin reductase regeneration system revealed that reduction susceptibility depended on not only antibody class but also light chain type; the model further demonstrates that the trend in reducibility was identical to DTT reduction sensitivity following the order IgG1λ > IgG1κ > IgG2λ > IgG2κ. Thus, both product attributes and process parameters contribute to the extent of antibody reduction during production. PMID:23751615

  19. Overexpression of protein disulfide isomerase in Aspergillus.

    PubMed

    El-Adawi, H; Khanh, N Q; Gassen, H

    2000-10-01

    One of the major problems with the production of biotechnologically valuable proteins has been the purification of the product. For Escherichia coli and Saccharomyces cerevisiae, there are several techniques for the purification of intracellular proteins, but these are time consuming and often result in poor yields. Purification can be considerably facilitated, if the product is secreted from the host cell. In the work presented, we have constructed an expression vector (pSGNH2) for the secretion of protein disulfide isomerase (PDI; EC 5.3.4.1) from Aspergillus niger, in which the retention signal His-Asp-Glu-Leu (H-D-E-L) was modified to Ala-Leu-Glu-Gln (A-L-E-Q) via the polymerase chain reaction (PCR) method. The PDI gene was placed under the control of the A. oryzae alpha-amylase promoter. This expression vector was transformed into A. niger NRRL3, resulting in PDI secretion into the medium. The catalytic activity of overexpressed PDI from A. niger was indistinguishable from that of PDI isolated from bovine liver. With further strain improvement and optimization of culture conditions, it could be possible to raise the PDI production to the bioprocessing scale. PMID:10977899

  20. Design, Synthesis and Biological Evaluation of Brain-Targeted Thiamine Disulfide Prodrugs of Ampakine Compound LCX001.

    PubMed

    Xiao, Dian; Meng, Fan-Hua; Dai, Wei; Yong, Zheng; Liu, Jin-Qiu; Zhou, Xin-Bo; Li, Song

    2016-01-01

    Ampakine compounds have been shown to reverse opiate-induced respiratory depression by activation of amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptors. However, their pharmacological exploitations are hindered by low blood-brain barrier (BBB) permeability and limited brain distribution. Here, we explored whether thiamine disulfide prodrugs with the ability of "lock-in" can be used to solve these problems. A series of thiamine disulfide prodrugs 7a-7f of ampakine compound LCX001 was synthesized and evaluated. The trials in vitro showed that prodrugs 7e, 7d, 7f possessed a certain stability in plasma and quickly decomposed in brain homogenate by the disulfide reductase. In vivo, prodrug 7e decreased the peripheral distribution of LCX001 and significantly increased brain distribution of LCX001 after i.v. administration. This compound showed 2.23- and 3.29-fold greater increases in the AUC0-t and MRT0-t of LCX001 in brain, respectively, than did LCX001 itself. A preliminary pharmacodynamic study indicated that the required molar dose of prodrug 7e was only one eighth that of LCX001 required to achieve the same effect in mice. These findings provide an important reference to evaluate the clinical outlook of ampakine compounds. PMID:27089316

  1. Rhenium Disulfide Depletion-Load Inverter

    NASA Astrophysics Data System (ADS)

    McClellan, Connor; Corbet, Chris; Rai, Amritesh; Movva, Hema C. P.; Tutuc, Emanuel; Banerjee, Sanjay K.

    2015-03-01

    Many semiconducting Transition Metal Dichalcogenide (TMD) materials have been effectively used to create Field-Effect Transistor (FET) devices but have yet to be used in logic designs. We constructed a depletion-load voltage inverter using ultrathin layers of Rhenium Disulfide (ReS2) as the semiconducting channel. This ReS2 inverter was fabricated on a single micromechanically-exfoliated flake of ReS2. Electron beam lithography and physical vapor deposition were used to construct Cr/Au electrical contacts, an Alumina top-gate dielectric, and metal top-gate electrodes. By using both low (Aluminum) and high (Palladium) work-function metals as two separate top-gates on a single ReS2 flake, we create a dual-gated depletion mode (D-mode) and enhancement mode (E-mode) FETs in series. Both FETs displayed current saturation in the output characteristics as a result of the FET ``pinch-off'' mechanism and On/Off current ratios of 105. Field-effect mobilities of 23 and 17 cm2V-1s-1 and subthreshold swings of 97 and 551 mV/decade were calculated for the E-mode and D-mode FETs, respectively. With a supply voltage of 1V, at low/negative input voltages the inverter output was at a high logic state of 900 mV. Conversely with high/positive input voltages, the inverter output was at a low logic state of 500 mV. The inversion of the input signal demonstrates the potential for using ReS2 in future integrated circuit designs and the versatility of depletion-load logic devices for TMD research. NRI SWAN Center and ARL STTR Program.

  2. AMU NEXRAD Exploitation Task

    NASA Technical Reports Server (NTRS)

    Lambert, Winifred C.; Wheeler, Mark M.

    1997-01-01

    This report documents the results of the Applied Meteorology Unit's NEXRAD Exploitation Task. The objectives of this task are to determine what radar signatures are present prior to and at the time of convection initiation, and to determine radar signatures which will help distinguish whether the ensuing convection will become severe. Radar data from the WSR-88D radar located at NWS Melbourne (WSR-88D/KMLB) were collected between June and September 1995, and 16 convective case studies were analyzed for which the radar was operating during the entire period of interest. All WSR-88D/KMLB products were scrutinized for their utility in detecting convection initiation and severe storm signatures. Through process of elimination, it was found that the 0.5 deg reflectivity product with the lowest reflectivity values displayed is the best product to monitor for convection initiation signatures. Seven meteorological features associated with the initiation of deep convection were identified: the Merritt Island and Indian River convergence zones, interlake convergence, horizontal convective rolls, the sea breeze, storm outflow boundaries, and fires. Their reflectivity values ranged from -5 to 20 dBZ. Of the three severe weather phenomena (winds greater than or equal to 50 kts, tornado, 3/4 inch hail), high wind events due to microbursts were most common in the data set. It was found that the values and trends of composite reflectivity, vertically integrated liquid, and core aspect ratio were key indicators of the potential of a cell to produce a microburst. The data were not analyzed for the other two severe weather phenomena because they rarely occurred during the data collection period. This report also includes suggestions for new WSR-88D products, summaries of ongoing research aimed at creating new products, and explicit recommended procedures for detecting convection initiation and severe storm signatures in the radar data using the currently available technology.

  3. Engineering Knowledge for Assistive Living

    NASA Astrophysics Data System (ADS)

    Chen, Liming; Nugent, Chris

    This paper introduces a knowledge based approach to assistive living in smart homes. It proposes a system architecture that makes use of knowledge in the lifecycle of assistive living. The paper describes ontology based knowledge engineering practices and discusses mechanisms for exploiting knowledge for activity recognition and assistance. It presents system implementation and experiments, and discusses initial results.

  4. Thiol/disulfide redox states in signaling and sensing

    PubMed Central

    Go, Young-Mi; Jones, Dean P.

    2015-01-01

    Rapid advances in redox systems biology are creating new opportunities to understand complexities of human disease and contributions of environmental exposures. New understanding of thiol-disulfide systems have occurred during the past decade as a consequence of the discoveries that thiol and disulfide systems are maintained in kinetically controlled steady-states displaced from thermodynamic equilibrium, that a widely distributed family of NADPH oxidases produces oxidants that function in cell signaling, and that a family of peroxiredoxins utilize thioredoxin as a reductant to complement the well-studied glutathione antioxidant system for peroxide elimination and redox regulation. This review focuses on thiol/disulfide redox state in biologic systems and the knowledge base available to support development of integrated redox systems biology models to better understand the function and dysfunction of thiol-disulfide redox systems. In particular, central principles have emerged concerning redox compartmentalization and utility of thiol/disulfide redox measures as indicators of physiologic function. Advances in redox proteomics show that, in addition to functioning in protein active sites and cell signaling, cysteine residues also serve as redox sensors to integrate biologic functions. These advances provide a framework for translation of redox systems biology concepts to practical use in understanding and treating human disease. Biological responses to cadmium, a widespread environmental agent, are used to illustrate the utility of these advances to the understanding of complex pleiotropic toxicities. PMID:23356510

  5. Does Electron Capture Dissociation Cleave Protein Disulfide Bonds?

    PubMed Central

    Ganisl, Barbara; Breuker, Kathrin

    2012-01-01

    Peptide and protein characterization by mass spectrometry (MS) relies on their dissociation in the gas phase into specific fragments whose mass values can be aligned as ‘mass ladders’ to provide sequence information and to localize possible posttranslational modifications. The most common dissociation method involves slow heating of even-electron (M+n H)n+ ions from electrospray ionization by energetic collisions with inert gas, and cleavage of amide backbone bonds. More recently, dissociation methods based on electron capture or transfer were found to provide far more extensive sequence coverage through unselective cleavage of backbone N–Cα bonds. As another important feature of electron capture dissociation (ECD) and electron transfer dissociation (ETD), their unique unimolecular radical ion chemistry generally preserves labile posttranslational modifications such as glycosylation and phosphorylation. Moreover, it was postulated that disulfide bond cleavage is preferred over backbone cleavage, and that capture of a single electron can break both a backbone and a disulfide bond, or even two disulfide bonds between two peptide chains. However, the proposal of preferential disulfide bond cleavage in ECD or ETD has recently been debated. The experimental data presented here reveal that the mechanism of protein disulfide bond cleavage is much more intricate than previously anticipated. PMID:24363980

  6. Thiol/disulfide homeostasis in patients with ankylosing spondylitis

    PubMed Central

    Dogru, Atalay; Balkarli, Ayse; Cetin, Gozde Yildirim; Neselioglu, Salim; Erel, Ozcan; Tunc, Sevket Ercan; Sahin, Mehmet

    2016-01-01

    Ankylosing spondylitis (AS) is a chronic inflammatory disease. In many inflammatory diseases, increased production of pro-inflammatory cytokines is associated with an increase in oxidative stress mediators. Thiol/disulfide homeostasis is a marker for oxidative stress. The aim of this study was to examine the dynamic thiol/disulfide homeostasis in AS. Sixty-nine patients with AS and 60 age- and sex-matched controls were included in the study. The Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and visual analogue scale (VAS) were used to determine the disease activity. Native thiol, total thiol, and disulfide levels were measured with a novel automated method recently described by Erel and Neselioglu. The aforementioned method is also optionally manual spectrophotometric assay. The total thiol levels were significantly lower in the AS group compared with the control group (p = 0.03). When the patients were divided into active (n = 35) and inactive (n = 34) subgroups using BASDAI scores, the native plasma thiol and total thiol levels were significantly lower in the active AS patients compared to the inactive AS patients (p = 0.02, p = 0.03 respectively). There was a negative correlation between the plasma native thiol levels and VAS, BASDAI scores. Thiol/disulfide homeostasis may be used for elucidating the effects of oxidative stress in AS. Understanding the role of thiol/disulfide homeostasis in AS might provide new therapeutic intervention strategies for patients.

  7. Disulfide Bridges: Bringing Together Frustrated Structure in a Bioactive Peptide.

    PubMed

    Zhang, Yi; Schulten, Klaus; Gruebele, Martin; Bansal, Paramjit S; Wilson, David; Daly, Norelle L

    2016-04-26

    Disulfide bridges are commonly found covalent bonds that are usually believed to maintain structural stability of proteins. Here, we investigate the influence of disulfide bridges on protein dynamics through molecular dynamics simulations on the cysteine-rich trypsin inhibitor MCoTI-II with three disulfide bridges. Correlation analysis of the reduced cyclic peptide shows that two of the three disulfide distances (Cys(11)-Cys(23) and Cys(17)-Cys(29)) are anticorrelated within ∼1 μs of bridge formation or dissolution: when the peptide is in nativelike structures and one of the distances shortens to allow bond formation, the other tends to lengthen. Simulations over longer timescales, when the denatured state is less structured, do not show the anticorrelation. We propose that the native state contains structural elements that frustrate one another's folding, and that the two bridges are critical for snapping the frustrated native structure into place. In contrast, the Cys(4)-Cys(21) bridge is predicted to form together with either of the other two bridges. Indeed, experimental chromatography and nuclear magnetic resonance data show that an engineered peptide with the Cys(4)-Cys(21) bridge deleted can still fold into its near-native structure even in its noncyclic form, confirming the lesser role of the Cys(4)-Cys(21) bridge. The results highlight the importance of disulfide bridges in a small bioactive peptide to bring together frustrated structure in addition to maintaining protein structural stability. PMID:27119635

  8. Inhibition of botulinum neurotoxins interchain disulfide bond reduction prevents the peripheral neuroparalysis of botulism.

    PubMed

    Zanetti, Giulia; Azarnia Tehran, Domenico; Pirazzini, Marcon; Binz, Thomas; Shone, Clifford C; Fillo, Silvia; Lista, Florigio; Rossetto, Ornella; Montecucco, Cesare

    2015-12-01

    Botulinum neurotoxins (BoNTs) form a growing family of metalloproteases with a unique specificity either for VAMP, SNAP25 or syntaxin. The BoNTs are grouped in seven different serotypes indicated by letters from A to G. These neurotoxins enter the cytosol of nerve terminals via a 100 kDa chain which binds to the presynaptic membrane and assists the translocation of a 50 kDa metalloprotease chain. These two chains are linked by a single disulfide bridge which plays an essential role during the entry of the metalloprotease chain in the cytosol, but thereafter it has to be reduced to free the proteolytic activity. Its reduction is mediated by thioredoxin which is continuously regenerated by its reductase. Here we show that inhibitors of thioredoxin reductase or of thioredoxin prevent the specific proteolysis of VAMP by the four VAMP-specific BoNTs: type B, D, F and G. These compounds are effective not only in primary cultures of neurons, but also in preventing the in vivo mouse limb neuroparalysis. In addition, one of these inhibitors, Ebselen, largely protects mice from the death caused by a systemic injection. Together with recent results obtained with BoNTs specific for SNAP25 and syntaxin, the present data demonstrate the essential role of the thioredoxin-thioredoxin reductase system in reducing the interchain disulfide during the nerve intoxication mechanism of all serotypes. Therefore its inhibitors should be considered for a possible use to prevent botulism and for treating infant botulism. PMID:26449594

  9. Disulfide Bond Formation in Prokaryotes: History, Diversity and Design

    PubMed Central

    Hatahet, Feras; Boyd, Dana; Beckwith, Jon

    2014-01-01

    The formation of structural disulfide bonds is essential for the function and stability of a great number of proteins, particularly those that are secreted. There exists a variety of dedicated cellular catalysts and pathways from Archaea to humans that ensure the formation of native disulfide bonds. In this review we describe the initial discoveries of these pathways and report progress in recent years in our understanding of the diversity of these pathways in prokaryotes, including those newly discovered in some Archaea. We will also discuss the various successful efforts to achieve laboratory-based evolution and design of synthetic disulfide bond formation machineries in the bacterium E. coli. These latter studies have also led to new more general insights into the redox environment of the cytoplasm and bacterial cell envelope. PMID:24576574

  10. Identification of disulfides from the biodegradation of dibenzothiophene.

    PubMed

    Bressler, D C; Fedorak, P M

    2001-11-01

    Several investigations have identified benzothiophene-2,3-dione in the organic solvent extracts of acidified cultures degrading dibenzothiophene via the Kodama pathway. In solution at neutral pH, the 2,3-dione exists as 2-mercaptophenylglyoxylate, which cyclizes upon acidification and is extracted as the 2,3-dione. The fate of these compounds in microbial cultures has never been determined. This study investigated the abiotic reactions of 2-mercaptophenylglyoxylate incubated aerobically in mineral salts medium at neutral pH. Oxidation led to the formation of 2-oxo-2-(2-thiophenyl)ethanoic acid disulfide, formed from two molecules of 2-mercaptophenylglyoxylate. Two sequential abiotic, net losses of both a carbon and an oxygen atom produced two additional disulfides, 2-oxo-2-(2-thiophenyl)ethanoic acid 2-benzoic acid disulfide and 2,2'-dithiosalicylic acid. The methods developed to extract and detect these three disulfides were then used for the analysis of a culture of Pseudomonas sp. strain BT1d grown on dibenzothiophene as its sole carbon and energy source. All three of the disulfides were detected, indicating that 2-mercaptophenylglyoxylate is an important, short-lived intermediate in the breakdown of dibenzothiophene via the Kodama pathway. The disulfides eluded previous investigations because of (i) their high polarity, being dicarboxylic acids; (ii) the need to lower the pH of the aqueous medium to <1 to extract them into an organic solvent such as dichloromethane; (iii) their poor solubility in organic solvents, (iv) their removal from organic extracts of cultures during filtration through the commonly used drying agent anhydrous sodium sulfate; and (v) their high molecular masses (362, 334, and 306 Da) compared to that of dibenzothiophene (184 Da). PMID:11679330

  11. Structures and related properties of helical, disulfide-stabilized peptides

    SciTech Connect

    Pagel, M.D. |

    1993-11-01

    The three dimensional structure of several peptides were determined by NMR spectroscopy and distance geometry calculations. Each peptide formed a predictable, rigid structure, consisting of an {alpha}-helix, a {open_quotes}scaffold{close_quotes} region which packed along one face of the helix, and two disulfide bridges which covalently connect the helix and scaffold regions. The peptide Apa-M5 was designed to constrain the M5 peptide from MLCK in a helical geometry using the apamin disulfide scaffold. This scaffold constrains the N- terminal end of the helix with two disulfide bridges and a reverse turn. Like the M5 peptide, Apa-M5 was found to bind calmodulin in a Ca{sup 2+}-dependent 1:1 stoichiometry. However, the dissociation constant of the (Apa-M5)-calmodulin complex, 107 nM, was 100-fold higher than the dissociation constant of the M5-calmodulin complex. This difference was due to a putative steric overlap between the Apa-M5 scaffold and calmodulin. The peptide Apa-Cro was designed to replace the large structural protein matrix of {lambda} Cro with the apamin disulfide scaffold. However, Apa-Cro did not bind the consensus DNA operator half-site of {lambda} Cro, probably due to a steric overlap between the Apa-Cro disulfide framework and the DNA. The amino acid sequence of the scaffold-disulfide bridge arrangement of the peptide Max was derived from the core sequence of scyllatoxin, which contains an {alpha}-helix constrained at the C-terminal end by two disulfide bridges and a two-stranded {beta}sheet scaffold. Max was shown to fold with >84% yield to form a predictable, stable structure that is similar to scyllatoxin. The folding and stability properties of Max make this scaffold and disulfide bridge arrangement an ideal candidate for the development of hybrid sequence peptides. The dynamics of a fraying C-terminal end of the helix of the peptide Apa-AlaN was determined by analysis of {sup 15}N NMR relaxation properties.

  12. Electrochemical study of thiols and disulfides using modified electrodes

    SciTech Connect

    Linders, C.R.; Patriarche, G.J.; Kauffman, J.M.

    1986-01-01

    The electrochemical oxidative behavior of cysteine and several disulfides, such as cysteine, lipoic acid and disulfiram, have been investigated using a carbon paste (EPC) and a modified carbon paste (EPCM) electrode. The study has permitted the differentiation of the oxidative behavior of the thiol and of the disulfides. Modification of the carbon paste, by incorporating cobalt(II) phthalocyanine, offers interesting properties due to the electrocatalytic capability of the electrode. Using these types of electrodes the different molecules have been quantitatively determined at concentrations as low as 2.10/sup -7/ M. 14 references, 2 figures, 1 table.

  13. Reduction of the lipocalin type heme containing protein nitrophorin -- sensitivity of the fold-stabilizing cysteine disulfides toward routine heme-iron reduction.

    PubMed

    Knipp, Markus; Taing, Johanna J; He, Chunmao

    2011-11-01

    The determination of the redox properties of the cofactor in heme proteins provides fundamental insight into the chemical characteristics of this wide-spread class of metalloproteins. For the preparation of the ferroheme state, probably the most widely applied reductant is sodium dithionite, which at neutral pH has a reduction potential well below the reduction potential of most heme centers. In addition to the heme iron, some heme proteins, including the nitrophorins (NPs), contain cysteinecysteine disulfide bonds. In the present study, the effect of dithionite on the disulfides of NP4 and NP7 is addressed. To gain deeper understanding of the disulfide/dithionite reaction, oxidized glutathione (GSSG), as a model system, was incubated with dithionite and the products were characterized by (13)C NMR spectroscopy and reverse phase chromatography in combination with mass spectrometry. This revealed the formation of one equivalent each of thiol (GSH) and glutathione-S-thiosulfate (GSSO(3)(-)). With this background information, the effect of dithionite on the cystines of NP4 and NP7 was studied after trapping of the thiols with para-cloromercurybenzyl sulfonate (p-CMBS) and subsequent matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) where the heterolytic cleavage of the SS bond appears with only 2molar equivalents of the reductant. Furthermore, prolonged electrochemical reduction of NP4 and NP7 in the presence of electrochemical mediators also leads to disulfide breakage. However, due to sterical shielding of the disulfide bridges in NP4 and NP7, the cystine reduction can be largely prevented by the use of stoichiometric amounts of reductant or limited electrochemical reduction. The described disulfide breakage during routine iron reduction is of importance for other heme proteins containing cystine(s). PMID:21955842

  14. Synergistic toughening of graphene oxide-molybdenum disulfide-thermoplastic polyurethane ternary artificial nacre.

    PubMed

    Wan, Sijie; Li, Yuchen; Peng, Jingsong; Hu, Han; Cheng, Qunfeng; Jiang, Lei

    2015-01-27

    Inspired by the ternary structure of natural nacre, robust ternary artificial nacre is constructed through synergistic toughening of graphene oxide (GO) and molybdenum disulfide (MoS2) nanosheets via a vacuum-assisted filtration self-assembly process. The synergistic toughening effect from high mechanical properties of GO and lubrication of MoS2 nanosheets is successfully demonstrated. Meanwhile, the artificial nacre shows high electrical conductivity. This approach for constructing robust artificial nacre by synergistic effect from GO and MoS2 provides a creative opportunity for designing and fabricating integrated artificial nacre in the near future, and this kind of ternary artificial nacre has great potential applications in aerospace, flexible supercapacitor electrodes, artificial muscle, and tissue engineering. PMID:25559751

  15. 21 CFR 520.1802b - Piperazine-carbon disulfide complex boluses.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Piperazine-carbon disulfide complex boluses. 520....1802b Piperazine-carbon disulfide complex boluses. (a) Specifications. Each bolus contains 20 grams of piperazine-carbon disulfide complex. (b) Sponsor. See 000009 in § 510.600(c) of this chapter. (c)...

  16. 21 CFR 520.1802c - Piperazine-carbon disulfide complex with phenothiazine suspension.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Piperazine-carbon disulfide complex with... ANIMAL DRUGS § 520.1802c Piperazine-carbon disulfide complex with phenothiazine suspension. (a) Specifications. Each fluid ounce contains 5 grams of piperazine-carbon disulfide complex and 0.83 gram...

  17. 21 CFR 520.1802b - Piperazine-carbon disulfide complex boluses.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Piperazine-carbon disulfide complex boluses. 520....1802b Piperazine-carbon disulfide complex boluses. (a) Specifications. Each bolus contains 20 grams of piperazine-carbon disulfide complex. (b) Sponsor. See 000009 in § 510.600(c) of this chapter. (c)...

  18. 21 CFR 520.1802c - Piperazine-carbon disulfide complex with phenothiazine suspension.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Piperazine-carbon disulfide complex with... ANIMAL DRUGS § 520.1802c Piperazine-carbon disulfide complex with phenothiazine suspension. (a) Specifications. Each fluid ounce contains 5 grams of piperazine-carbon disulfide complex and 0.83 gram...

  19. 21 CFR 520.1802b - Piperazine-carbon disulfide complex boluses.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Piperazine-carbon disulfide complex boluses. 520....1802b Piperazine-carbon disulfide complex boluses. (a) Specifications. Each bolus contains 20 grams of piperazine-carbon disulfide complex. (b) Sponsor. See 000009 in § 510.600(c) of this chapter. (c)...

  20. 21 CFR 520.1802c - Piperazine-carbon disulfide complex with phenothiazine suspension.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Piperazine-carbon disulfide complex with... ANIMAL DRUGS § 520.1802c Piperazine-carbon disulfide complex with phenothiazine suspension. (a) Specifications. Each fluid ounce contains 5 grams of piperazine-carbon disulfide complex and 0.83 gram...

  1. 21 CFR 520.1802b - Piperazine-carbon disulfide complex boluses.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Piperazine-carbon disulfide complex boluses. 520....1802b Piperazine-carbon disulfide complex boluses. (a) Specifications. Each bolus contains 20 grams of piperazine-carbon disulfide complex. (b) Sponsor. See 000009 in § 510.600(c) of this chapter. (c)...

  2. 21 CFR 520.1802c - Piperazine-carbon disulfide complex with phenothiazine suspension.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Piperazine-carbon disulfide complex with... ANIMAL DRUGS § 520.1802c Piperazine-carbon disulfide complex with phenothiazine suspension. (a) Specifications. Each fluid ounce contains 5 grams of piperazine-carbon disulfide complex and 0.83 gram...

  3. 21 CFR 520.1802c - Piperazine-carbon disulfide complex with phenothiazine suspension.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Piperazine-carbon disulfide complex with... ANIMAL DRUGS § 520.1802c Piperazine-carbon disulfide complex with phenothiazine suspension. (a) Specifications. Each fluid ounce contains 5 grams of piperazine-carbon disulfide complex and 0.83 gram...

  4. 21 CFR 520.1802b - Piperazine-carbon disulfide complex boluses.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Piperazine-carbon disulfide complex boluses. 520....1802b Piperazine-carbon disulfide complex boluses. (a) Specifications. Each bolus contains 20 grams of piperazine-carbon disulfide complex. (b) Sponsor. See 000009 in § 510.600(c) of this chapter. (c)...

  5. Conserved residues flanking the thiol/disulfide centers of protein disulfide isomerase are not essential for catalysis of thiol/disulfide exchange.

    PubMed

    Lu, X; Gilbert, H F; Harper, J W

    1992-05-01

    Protein disulfide isomerase (PDI) catalyzes the oxidative folding of proteins containing disulfide bonds by increasing the rate of disulfide bond rearrangements which normally occur during the folding process. The amino acid sequences of the N- and C-terminal redox active sites (PWCGHCK) in PDI are completely conserved from yeast to man and display considerable identity with the redox-active center of thioredoxin (EWCGPCK). Available data indicate that the two thiol/disulfide centers of PDI can function independently in the isomerase reaction and that the cysteine residues in each active site are essential for catalysis. To evaluate the role of residues flanking the active-site cysteines of PDI in function, a variety of mutations were introduced into the N-terminal active site of PDI within the context of both a functional C-terminal active site and an inactive C-terminal active site in which serine residues replaced C379 and C382. Replacement of non-cysteine residues (W34 to Ser, G36 to Ala, and K39 to Arg) resulted in only a modest reduction in catalytic activity in both the oxidative refolding of RNase A and the reduction of insulin (10-27%), independent of the status of the C-terminal active site. A somewhat larger effect was observed with the H37P mutation where approximately 80% of the activity attributable to the N-terminal domain (approximately 40%) was lost. However, the H37P mutant N-terminal site expressed within the context of an inactive C-terminal domain exhibits 30% activity, approximately 70% of the activity of the N-terminal site alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1567868

  6. Response of soil organisms to dimethyl disulfide fumigation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    After the commonly used soil fumigant methyl bromide (MeBr) was phased out in the United States, alternatives to MeBr such as dimethyl disulfide (DMDS) which is known to have broad pest control spectrum, is increasingly used. However, effectiveness of DMDS has been mainly investigated to study targe...

  7. HEALTH AND ENVIRONMENTAL EFFECTS PROFILE FOR CARBON DISULFIDE

    EPA Science Inventory

    The Health and Environmental Effects Profile for Carbon Disulfide was prepared to support listings of hazardous constituents of a wide range of waste streams under Section 3001 of the Resource Conservation and Recovery Act (RCRA) and to provide health-related limits for emergency...

  8. Application of the solid lubricant molybdenum disulfide by sputtering

    NASA Technical Reports Server (NTRS)

    Przybyszewski, J.; Spalvins, T.

    1968-01-01

    Molybdenum disulfide lubricant film is deposited on two substrates, niobium and nickel-chromium alloys, by means of physical direct-current sputtering. The sputtering system uses a three-electrode /triode/ geometry - a thermionic cathode, an anode, and the target, all enclosed in a vacuum chamber.

  9. A kinetic study on pantetheinase inhibition by disulfides.

    PubMed

    Pitari, G; Maurizi, G; Ascenzi, P; Ricci, G; Duprè, S

    1994-11-15

    The mammalian enzyme pantetheinase, which hydrolyzes pantetheine to pantothenic acid and cysteamine, is inhibited by many thiol reagents and activated by thiols. Two thiol groups of different reactivity and accessibility are involved in the catalytic process [Ricci, G., Nardini, M., Chiaraluce, R., Duprè, S. & Cavallini, D. (1986) Biochim. Biophys. Acta 870, 82-91]. The inhibition kinetics by some natural and synthetic disulfides [pantethine, cystamine, 5,5'-dithiobis(2-nitrobenzoic acid), 4,4'-dithiodipyridine and oxidized mercaptoethanol] has been studied by two experimental approaches, either by monitoring activity after incubation of the enzyme with the inhibitor or by determining the progress curves in the presence of substrate and inhibitor. Data reported here indicate that pantetheinase reacts irreversibly with various disulfides in a time-dependent manner with the formation of a mixed disulfide apparently preceeded by a conformational change, giving a modified E* form with new kinetic parameters. This modified form may be further competitively inhibited by disulfides interacting with the enzyme at the active site. PMID:7957261

  10. Disulfide-based multifunctional conjugates for targeted theranostic drug delivery.

    PubMed

    Lee, Min Hee; Sessler, Jonathan L; Kim, Jong Seung

    2015-11-17

    Theranostics, chemical entities designed to combine therapeutic effects and imaging capability within one molecular system, have received considerable attention in recent years. Much of this interest reflects the promise inherent in personalized medicine, including disease-targeted treatments for cancer patients. One important approach to realizing this latter promise involves the development of so-called theranostic conjugates, multicomponent constructs that selectively target cancer cells and deliver cytotoxic agents while producing a readily detectable signal that can be monitored both in vitro and in vivo. This requires the synthesis of relatively complex systems comprising imaging reporters, masked chemotherapeutic drugs, cleavable linkers, and cancer targeting ligands. Ideally, the cleavage process should take place within or near cancer cells and be activated by cellular components that are associated with cancer states or specifically expressed at a higher level in cancer cells. Among the cleavable linkers currently being explored for the construction of such localizing conjugates, disulfide bonds are particularly attractive. This is because disulfide bonds are stable in most blood pools but are efficiently cleaved by cellular thiols, including glutathione (GSH) and thioredoxin (Trx), which are generally found at elevated levels in tumors. When disulfide bonds are linked to fluorophores, changes in emission intensity or shifts in the emission maxima are typically seen upon cleavage as the result of perturbations to internal charge transfer (ICT) processes. In well-designed systems, this allows for facile imaging. In this Account, we summarize our recent studies involving disulfide-based fluorescent drug delivery conjugates, including preliminary tests of their biological utility in vitro and in vivo. To date, a variety of chemotherapeutic agents, such as doxorubicin, gemcitabine, and camptothecin, have been used to create disulfide-based conjugates, as have

  11. Coenzyme A disulfide reductase, the primary low molecular weight disulfide reductase from Staphylococcus aureus. Purification and characterization of the native enzyme.

    PubMed

    delCardayre, S B; Stock, K P; Newton, G L; Fahey, R C; Davies, J E

    1998-03-01

    The human pathogen Staphylococcus aureus does not utilize the glutathione thiol/disulfide redox system employed by eukaryotes and many bacteria. Instead, this organism produces CoA as its major low molecular weight thiol. We report the identification and purification of the disulfide reductase component of this thiol/disulfide redox system. Coenzyme A disulfide reductase (CoADR) catalyzes the specific reduction of CoA disulfide by NADPH. CoADR has a pH optimum of 7.5-8.0 and is a dimer of identical subunits of Mr 49,000 each. The visible absorbance spectrum is indicative of a flavoprotein with a lambdamax = 452 nm. The liberated flavin from thermally denatured enzyme was identified as flavin adenine dinucleotide. Steady-state kinetic analysis revealed that CoADR catalyzes the reduction of CoA disulfide by NADPH at pH 7.8 with a Km for NADPH of 2 muM and for CoA disulfide of 11 muM. In addition to CoA disulfide CoADR reduces 4,4'-diphosphopantethine but has no measurable ability to reduce oxidized glutathione, cystine, pantethine, or H2O2. CoADR demonstrates a sequential kinetic mechanism and employs a single active site cysteine residue that forms a stable mixed disulfide with CoA during catalysis. These data suggest that S. aureus employs a thiol/disulfide redox system based on CoA/CoA-disulfide and CoADR, an unorthodox new member of the pyridine nucleotide-disulfide reductase superfamily. PMID:9488707

  12. Mitochondrial Ccs1 contains a structural disulfide bond crucial for the import of this unconventional substrate by the disulfide relay system

    PubMed Central

    Groß, Dominik P.; Burgard, Caroline A.; Reddehase, Silvia; Leitch, Jeffry M.; Culotta, Valeria C.; Hell, Kai

    2011-01-01

    The copper chaperone for superoxide dismutase 1 (Ccs1) provides an important cellular function against oxidative stress. Ccs1 is present in the cytosol and in the intermembrane space (IMS) of mitochondria. Its import into the IMS depends on the Mia40/Erv1 disulfide relay system, although Ccs1 is, in contrast to typical substrates, a multidomain protein and lacks twin CxnC motifs. We report on the molecular mechanism of the mitochondrial import of Saccharomyces cerevisiae Ccs1 as the first member of a novel class of unconventional substrates of the disulfide relay system. We show that the mitochondrial form of Ccs1 contains a stable disulfide bond between cysteine residues C27 and C64. In the absence of these cysteines, the levels of Ccs1 and Sod1 in mitochondria are strongly reduced. Furthermore, C64 of Ccs1 is required for formation of a Ccs1 disulfide intermediate with Mia40. We conclude that the Mia40/Erv1 disulfide relay system introduces a structural disulfide bond in Ccs1 between the cysteine residues C27 and C64, thereby promoting mitochondrial import of this unconventional substrate. Thus the disulfide relay system is able to form, in addition to double disulfide bonds in twin CxnC motifs, single structural disulfide bonds in complex protein domains. PMID:21865601

  13. Nonnative Disulfide Bond Formation Activates the σ32-Dependent Heat Shock Response in Escherichia coli

    PubMed Central

    Müller, Alexandra; Hoffmann, Jörg H.; Meyer, Helmut E.; Narberhaus, Franz; Jakob, Ursula

    2013-01-01

    Formation of nonnative disulfide bonds in the cytoplasm, so-called disulfide stress, is an integral component of oxidative stress. Quantification of the extent of disulfide bond formation in the cytoplasm of Escherichia coli revealed that disulfide stress is associated with oxidative stress caused by hydrogen peroxide, paraquat, and cadmium. To separate the impact of disulfide bond formation from unrelated effects of these oxidative stressors in subsequent experiments, we worked with two complementary approaches. We triggered disulfide stress either chemically by diamide treatment of cells or genetically in a mutant strain lacking the major disulfide-reducing systems TrxB and Gor. Studying the proteomic response of E. coli exposed to disulfide stress, we found that intracellular disulfide bond formation is a particularly strong inducer of the heat shock response. Real-time quantitative PCR experiments showed that disulfide stress induces the heat shock response in E. coli σ32 dependently. However, unlike heat shock treatment, which induces these genes transiently, transcripts of σ32-dependent genes accumulated over time in disulfide stress-treated cells. Analyzing the stability of σ32, we found that this constant induction can be attributed to an increase of the half-life of σ32 upon disulfide stress. This is concomitant with aggregation of E. coli proteins treated with diamide. We conclude that oxidative stress triggers the heat shock response in E. coli σ32 dependently. The component of oxidative stress responsible for the induction of heat shock genes is disulfide stress. Nonnative disulfide bond formation in the cytoplasm causes protein unfolding. This stabilizes σ32 by preventing its DnaK- and FtsH-dependent degradation. PMID:23585533

  14. Sensory exploitation and sexual conflict

    PubMed Central

    Arnqvist, Göran

    2006-01-01

    Much of the literature on male–female coevolution concerns the processes by which male traits and female preferences for these can coevolve and be maintained by selection. There has been less explicit focus on the origin of male traits and female preferences. Here, I argue that it is important to distinguish origin from subsequent coevolution and that insights into the origin can help us appreciate the relative roles of various coevolutionary processes for the evolution of diversity in sexual dimorphism. I delineate four distinct scenarios for the origin of male traits and female preferences that build on past contributions, two of which are based on pre-existing variation in quality indicators among males and two on exploitation of pre-existing sensory biases among females. Recent empirical research, and theoretical models, suggest that origin by sensory exploitation has been widespread. I argue that this points to a key, but perhaps transient, role for sexually antagonistic coevolution (SAC) in the subsequent evolutionary elaboration of sexual traits, because (i) sensory exploitation is often likely to be initially costly for individuals of the exploited sex and (ii) the subsequent evolution of resistance to sensory exploitation should often be associated with costs due to selective constraints. A review of a few case studies is used to illustrate these points. Empirical data directly relevant to the costs of being sensory exploited and the costs of evolving resistance is largely lacking, and I stress that such data would help determining the general importance of sexual conflict and SAC for the evolution of sexual dimorphism. PMID:16612895

  15. A degradable polydopamine coating based on disulfide-exchange reaction.

    PubMed

    Hong, Daewha; Lee, Hojae; Kim, Beom Jin; Park, Taegyun; Choi, Ji Yu; Park, Matthew; Lee, Juno; Cho, Hyeoncheol; Hong, Seok-Pyo; Yang, Sung Ho; Jung, Sun Ho; Ko, Sung-Bo; Choi, Insung S

    2015-12-21

    Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies. PMID:26572596

  16. Evolution of interlayer coupling in twisted molybdenum disulfide bilayers

    NASA Astrophysics Data System (ADS)

    Liu, Kaihui; Zhang, Liming; Cao, Ting; Jin, Chenhao; Qiu, Diana; Zhou, Qin; Zettl, Alex; Yang, Peidong; Louie, Steve G.; Wang, Feng

    2014-09-01

    Van der Waals coupling is emerging as a powerful method to engineer physical properties of atomically thin two-dimensional materials. In coupled graphene-graphene and graphene-boron nitride layers, interesting physical phenomena ranging from Fermi velocity renormalization to Hofstadter’s butterfly pattern have been demonstrated. Atomically thin transition metal dichalcogenides, another family of two-dimensional-layered semiconductors, can show distinct coupling phenomena. Here we demonstrate the evolution of interlayer coupling with twist angles in as-grown molybdenum disulfide bilayers. We find that the indirect bandgap size varies appreciably with the stacking configuration: it shows the largest redshift for AA- and AB-stacked bilayers, and a significantly smaller but constant redshift for all other twist angles. Our observations, together with ab initio calculations, reveal that this evolution of interlayer coupling originates from the repulsive steric effects that leads to different interlayer separations between the two molybdenum disulfide layers in different stacking configurations.

  17. Pressure-time profile of multiply shocked carbon disulfide

    NASA Astrophysics Data System (ADS)

    Sutherland, G. T.; Gupta, Y. M.; Bellamy, P. M.

    1986-02-01

    An experimental method was developed to measure the pressure-time profile of a liquid in a reverberation of multiple-shock experiment. Profiles, with peak pressures to 30 kbars, were measured for carbon disulfide using shorted quartz gauges (25.4 mm diameter by 3.15 mm thick); these gauges formed the back surfaces of cells which contained the carbon disulfide. Sapphire plates were used both as impactors and as the front surfaces of the cell. Up to six pressure steps were clearly observed in the quartz-gauge output. Measured pressure-time profiles were compared to profiles calculated with available equations of state. The experiments agreed well with profiles predicted with an equation of state proposed by Sheffield (1983). Calibration experiments were performed to characterize both the initial current response and the subsequent current ramping of the shorted quartz gauges used in this study.

  18. Moderate temperature sodium cells. I - Transition metal disulfide cathodes

    NASA Technical Reports Server (NTRS)

    Abraham, K. M.; Pitts, L.; Schiff, R.

    1980-01-01

    TiS2, VS2, and Nb(1.1)S2 transition metal disulfides were evaluated as cathode materials for a moderate temperature rechargeable Na cell operating at 130 C. The 1st discharge of TiS2 results in a capacity of 0.85 eq/mole; approximately half of the Na in the 1st phase spanning the Na range from zero to 0.30 and almost all the Na in the 2nd phase spanning the 0.37 to 0.80 range are rechargeable. VS2 intercalates up to one mole of Na/mole of VS2 in the 1st discharge; the resulting Na(x)VS2 ternary consists of 3 phases in the 3 ranges of Na from zero to 1. Niobium disulfide undergoes a phase change in the 1st discharge; the average rechargeable capacity in extended cycling of this cathode is 0.50 eq/mole.

  19. Tibial dyschondroplasia in growing chickens experimentally intoxicated with tetramethylthiuram disulfide.

    PubMed

    Vargas, M I; Lamas, J M; Alvarenga, V

    1983-07-01

    Graded levels of tetramethylthiuram disulfide (0, 30, 60, 120, and 240 ppm) were incorporated into a broiler starter ration fed to chickens from one day old to 8 weeks of age. Clinical signs of leg abnormalities were observed as early as 5 days after the beginning of the trial. After the 3rd week, the joints were shown to present lesions, especially in the femorotibial articulation, comparable to the ones found in perosis. Tibiotarsus and other organs from the birds were examined for pathological changes at weekly intervals. Histologically, the tibiotarsus has shown an osteochondrodystrophy identical to that of tibial dyschondroplasia (TD), although it varied according to the level of tetramethylthiuram disulfide. The histopathology of the thyroid gland of the chickens involved in the present experiment will be reported in a separate manuscript. PMID:6622364

  20. High hemoglobin mixed disulfide content in hemolysates from stressed shark.

    PubMed

    Dafré, A L; Reischl, E

    1990-01-01

    1. Hemolysate from heavily stressed smooth hammerhead shark, Sphyrna zygaena, shows three electrophoretic components, SZ I, SZ II and SZ III, whose relative concentrations are 36.4 +/- 6.8, 36.4 +/- 5.0 and 20.8 +/- 5.7%, respectively. After reduction with DTE only SZ I remained. 2. SZ I reacted with glutathione disulfide reconstitute SZ II and SZ III. 3. Non-reduced, DTE-reduced, and denatured hemoglobin were found to have 2.0 +/- 0.4, 3.7 +/- 0.6, and 9.4 +/- 0.7-SH groups, respectively. 4. Erythrocyte non-protein--SH (NPSH), including glutathione present as mixed disulfide with SZ II and SZ III, is 1.7 NPSH/Hb. PMID:2361357

  1. A degradable polydopamine coating based on disulfide-exchange reaction

    NASA Astrophysics Data System (ADS)

    Hong, Daewha; Lee, Hojae; Kim, Beom Jin; Park, Taegyun; Choi, Ji Yu; Park, Matthew; Lee, Juno; Cho, Hyeoncheol; Hong, Seok-Pyo; Yang, Sung Ho; Jung, Sun Ho; Ko, Sung-Bo; Choi, Insung S.

    2015-11-01

    Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies.Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies. Electronic supplementary information (ESI) available: Synthesis, characterization, and other additional details. See DOI: 10

  2. Performance and Safety Characteristics of Lithium-molybdenum Disulfide Cells

    NASA Technical Reports Server (NTRS)

    Stiles, J. A.

    1984-01-01

    The lithium-molybdenum disulfide system offers attractive characteristics including high rate capability, successful operation up to 75 C, a very low self-discharge rate, a good cycle life and safety characteristics which compare favorably to those of other lithium cells. Moreover, the materials and manufacturing costs for the system is effectively controlled, so the cells should ultimately be competitive with currently marketed rechargeable cells.

  3. The Exploitation of Black Athletes.

    ERIC Educational Resources Information Center

    Edwards, Harry

    1983-01-01

    Colleges and universities have not up held their end of the bargain with athletes, exploiting a disproportionate number of talented Black athletes by not providing the kind of education the students sought or needed and by applying rigid academic standards for eligibility. (MSE)

  4. Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

    PubMed Central

    Ren, Guoping; Champion, Matthew M.; Huntley, Jason F.

    2014-01-01

    Summary Bacterial pathogens are exposed to toxic molecules inside the host and require efficient systems to form and maintain correct disulfide bonds for protein stability and function. The intracellular pathogen Francisella tularensis encodes a disulfide bond formation protein ortholog, DsbA, which previously was reported to be required for infection of macrophages and mice. However, the molecular mechanisms by which F. tularensis DsbA contributes to virulence are unknown. Here, we demonstrate that F. tularensis DsbA is a bifunctional protein that oxidizes and, more importantly, isomerizes complex disulfide connectivity in substrates. A single amino acid in the conserved cis-proline loop of the DsbA thioredoxin domain was shown to modulate both isomerase activity and F. tularensis virulence. Trapping experiments in F. tularensis identified over 50 F. tularensis DsbA substrates, including outer membrane proteins, virulence factors, and many hypothetical proteins. Six of these hypothetical proteins were randomly selected and deleted, revealing two novel proteins, FTL_1548 and FTL_1709, which are required for F. tularensis virulence. We propose that the extreme virulence of F. tularensis is partially due to the bifunctional nature of DsbA, that many of the newly-identified substrates are required for virulence, and that the development of future DsbA inhibitors could have broad anti-bacterial implications. PMID:25257164

  5. Polymerizable disulfide paclitaxel prodrug for controlled drug delivery.

    PubMed

    Ding, Yi; Chen, Wulian; Hu, Jianhua; Du, Ming; Yang, Dong

    2014-11-01

    A polymerizable disulfide paclitaxel (PTX) prodrug was synthesized by the consequential esterification reactions of 3,3'-dithiodipropionic acid (DTPA), a disulfide compound containing two active carboxyl groups, with 2-hydroxyethyl methacrylate (HEMA) and PTX. The structure of the prodrug was confirmed by (1)H NMR characterization. Then, the polymerizable prodrug was copolymerized with poly(ethylene glycol) methyl ether methacrylate (PEGMEA) to obtain a copolymer with hydrophilic PEG side chains and PTX covalently linked onto the backbone via disulfide bonds. The loading content of PTX was 23%. In aqueous solution, this copolymer prodrug could self-assemble into micelles, with hydrophobic PTX as the cores and hydrophilic PEG-segment as the shells. In vitro cell assay demonstrated that this copolymer prodrug showed more apparent cytotoxicity to cancer cells than to human normal cells. After incubation for 48 h, the cell viability of HEK-293 cells (human embryo kidney cells) at 0.1 μg/mL PTX still remained more than 90%, however, that of HeLa cells (human cervical cancer cells) decreased to 52%. PMID:25280719

  6. Dissecting the Machinery That Introduces Disulfide Bonds in Pseudomonas aeruginosa

    PubMed Central

    Arts, Isabelle S.; Ball, Geneviève; Leverrier, Pauline; Garvis, Steven; Nicolaes, Valérie; Vertommen, Didier; Ize, Bérengère; Tamu Dufe, Veronica; Messens, Joris; Voulhoux, Romé; Collet, Jean-François

    2013-01-01

    ABSTRACT Disulfide bond formation is required for the folding of many bacterial virulence factors. However, whereas the Escherichia coli disulfide bond-forming system is well characterized, not much is known on the pathways that oxidatively fold proteins in pathogenic bacteria. Here, we report the detailed unraveling of the pathway that introduces disulfide bonds in the periplasm of the human pathogen Pseudomonas aeruginosa. The genome of P. aeruginosa uniquely encodes two DsbA proteins (P. aeruginosa DsbA1 [PaDsbA1] and PaDsbA2) and two DsbB proteins (PaDsbB1 and PaDsbB2). We found that PaDsbA1, the primary donor of disulfide bonds to secreted proteins, is maintained oxidized in vivo by both PaDsbB1 and PaDsbB2. In vitro reconstitution of the pathway confirms that both PaDsbB1 and PaDsbB2 shuttle electrons from PaDsbA1 to membrane-bound quinones. Accordingly, deletion of both P. aeruginosa dsbB1 (PadsbB1) and PadsbB2 is required to prevent the folding of several P. aeruginosa virulence factors and to lead to a significant decrease in pathogenicity. Using a high-throughput proteomic approach, we also analyzed the impact of PadsbA1 deletion on the global periplasmic proteome of P. aeruginosa, which allowed us to identify more than 20 new potential substrates of this major oxidoreductase. Finally, we report the biochemical and structural characterization of PaDsbA2, a highly oxidizing oxidoreductase, which seems to be expressed under specific conditions. By fully dissecting the machinery that introduces disulfide bonds in P. aeruginosa, our work opens the way to the design of novel antibacterial molecules able to disarm this pathogen by preventing the proper assembly of its arsenal of virulence factors. PMID:24327342

  7. Teotihuacan, tepeapulco, and obsidian exploitation.

    PubMed

    Charlton, T H

    1978-06-16

    Current cultural ecological models of the development of civilization in central Mexico emphasize the role of subsistence production techniques and organization. The recent use of established and productive archeological surface survey techniques along natural corridors of communication between favorable niches for cultural development within the Central Mexican symbiotic region resulted in the location of sites that indicate an early development of a decentralized resource exploitation, manufacturing, and exchange network. The association of the development of this system with Teotihuacán indicates the importance such nonsubsistence production and exchange had in the evolution of this first central Mexican civilization. The later expansion of Teotihuacán into more distant areas of Mesoamerica was based on this resource exploitation model. Later civilizations centered at Tula and Tenochtitlán also used such a model in their expansion. PMID:17738704

  8. Exploiting perceptual redundancy in images

    NASA Astrophysics Data System (ADS)

    Liu, Hongyi; Chen, Zhenzhong

    2015-03-01

    Exploiting perceptual redundancy plays an important role in image processing. Conventional JND models describe the visibility of the minimally perceptible difference by assuming that the visual acuity is consistent over the whole image. Some earlier work considers the space-variant properties of HVS-based on the non-uniform density of photoreceptor cells. In this paper, we aim to exploit the relationship between the masking effects and the foveation properties of HVS. We design the psychophysical experiments which are conducted to model the foveation properties in response to the masking effects. The experiment examines the reduction of visual sensitivity in HVS due to the increased retinal eccentricity. Based on these experiments, the developed Foveated JND model measures the perceptible difference of images according to masking effects therefore provides the information to quantify the perceptual redundancy in the images. Subjective evaluations validate the proposed FJND model.

  9. Dark matters: exploitation as cooperation.

    PubMed

    Dasgupta, Partha

    2012-04-21

    The empirical literature on human cooperation contains studies of communitarian institutions that govern the provision of public goods and management of common property resources in poor countries. Scholars studying those institutions have frequently used the Prisoners' Dilemma game as their theoretical tool-kit. But neither the provision of local public goods nor the management of local common property resources involves the Prisoners' Dilemma. That has implications for our reading of communitarian institutions. By applying a fundamental result in the theory of repeated games to a model of local common property resources, it is shown that communitarian institutions can harbour exploitation of fellow members, something that would not be possible in societies where cooperation amounts to overcoming the Prisoners' Dilemma. The conclusion we should draw is that exploitation can masquerade as cooperation. PMID:21549130

  10. 25 CFR 20.516 - How are child abuse, neglect or exploitation cases to be handled?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... by the state will be handled according to the Indian Child Welfare Act, Public Law 95-608, and 25 CFR... 25 Indians 1 2013-04-01 2013-04-01 false How are child abuse, neglect or exploitation cases to be... FINANCIAL ASSISTANCE AND SOCIAL SERVICES PROGRAMS Child Assistance Foster Care § 20.516 How are child...

  11. 25 CFR 20.516 - How are child abuse, neglect or exploitation cases to be handled?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... by the state will be handled according to the Indian Child Welfare Act, Public Law 95-608, and 25 CFR... 25 Indians 1 2014-04-01 2014-04-01 false How are child abuse, neglect or exploitation cases to be... FINANCIAL ASSISTANCE AND SOCIAL SERVICES PROGRAMS Child Assistance Foster Care § 20.516 How are child...

  12. 25 CFR 20.516 - How are child abuse, neglect or exploitation cases to be handled?

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... by the state will be handled according to the Indian Child Welfare Act, Public Law 95-608, and 25 CFR... 25 Indians 1 2010-04-01 2010-04-01 false How are child abuse, neglect or exploitation cases to be... FINANCIAL ASSISTANCE AND SOCIAL SERVICES PROGRAMS Child Assistance Foster Care § 20.516 How are child...

  13. 25 CFR 20.516 - How are child abuse, neglect or exploitation cases to be handled?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... by the state will be handled according to the Indian Child Welfare Act, Public Law 95-608, and 25 CFR... 25 Indians 1 2012-04-01 2011-04-01 true How are child abuse, neglect or exploitation cases to be... FINANCIAL ASSISTANCE AND SOCIAL SERVICES PROGRAMS Child Assistance Foster Care § 20.516 How are child...

  14. 25 CFR 20.516 - How are child abuse, neglect or exploitation cases to be handled?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... by the state will be handled according to the Indian Child Welfare Act, Public Law 95-608, and 25 CFR... 25 Indians 1 2011-04-01 2011-04-01 false How are child abuse, neglect or exploitation cases to be... FINANCIAL ASSISTANCE AND SOCIAL SERVICES PROGRAMS Child Assistance Foster Care § 20.516 How are child...

  15. Cathode Based on Molybdenum Disulfide Nanoflakes for Lithium-Oxygen Batteries.

    PubMed

    Asadi, Mohammad; Kumar, Bijandra; Liu, Cong; Phillips, Patrick; Yasaei, Poya; Behranginia, Amirhossein; Zapol, Peter; Klie, Robert F; Curtiss, Larry A; Salehi-Khojin, Amin

    2016-02-23

    Lithium-oxygen (Li-O2) batteries have been recognized as an emerging technology for energy storage systems owing to their high theoretical specific energy. One challenge is to find an electrolyte/cathode system that is efficient, stable, and cost-effective. We present such a system based on molybdenum disulfide (MoS2) nanoflakes combined with an ionic liquid (IL) that work together as an effective cocatalyst for discharge and charge in a Li-O2 battery. Cyclic voltammetry results show superior catalytic performance for this cocatalyst for both oxygen reduction and evolution reactions compared to Au and Pt catalysts. It also performs remarkably well in the Li-O2 battery system with 85% round-trip efficiency and reversibility up to 50 cycles. Density functional calculations provide a mechanistic understanding of the MoS2 nanoflakes/IL system. The cocatalyst reported in this work could open the way for exploiting the unique properties of ionic liquids in Li-air batteries in combination with nanostructured MoS2 as a cathode material. PMID:26789516

  16. Formation of disulfide bonds in insect prophenoloxidase enhances immunity through improving enzyme activity and stability.

    PubMed

    Lu, Anrui; Peng, Qin; Ling, Erjun

    2014-06-01

    Type 3 copper proteins, including insect prophenoloxidase (PPO), contain two copper atoms in the active site pocket and can oxidize phenols. Insect PPO plays an important role in immunity. Insects and other invertebrates show limited recovery from pathogen invasion and wounds if phenoloxidase (PO) activity is low. In most insect PPOs, two disulfide bonds are present near the C-terminus. However, in Pimpla hypochondriaca (a parasitoid wasp), each PPO contains one disulfide bond. We thus questioned whether the formation of two sulfide bonds in insect PPOs improved protein stability and/or increased insect innate immunity over time. Using Drosophila melanogaster PPO1 as a model, one or two disulfide bonds were deleted to evaluate the importance of disulfide bonds in insect immunity. rPPO1 and mutants lacking disulfide bonds could be expressed and showed PO activity. However, the PO activities of mutants lacking one or two disulfide bonds significantly decreased. Deletion of disulfide bonds also reduced PPO thermostability. Furthermore, antibacterial activities against Escherichia coli and Bacillus subtilis significantly decreased when disulfide bonds were deleted. Therefore, the formation of two disulfide bond(s) in insect PPO enhances antibacterial activity by increasing PO activity and stability. PMID:24480295

  17. Method for removal of asphaltene depositions with amine-activated disulfide oil

    SciTech Connect

    Sharp, S.P.

    1983-04-12

    A method for treating and removing unwanted asphaltene deposits from oil and gas wells, surface equipment, flow lines, and pore spaces of oil-baring formations comprises treatment with an amine -activated aliphatic disulfide oil as an asphaltene solvent. In a preferred aspect, the aliphatic disulfide oil is a dialkyl disulfide oil and is activated by the addition of 10 weight percent of diethylamine. In a specific use, the activated disulfide oil is used to remove asphaltene deposits from an oilbearing formation and a producing well penetrating the formation.

  18. The influence of gold(i) on the mechanism of thiolate, disulfide exchange.

    PubMed

    Garusinghe, Gamage S P; Bessey, S Max; Bruce, Alice E; Bruce, Mitchell R M

    2016-07-28

    The mechanism of gold(i)-thiolate, disulfide exchange was investigated by using initial-rate kinetic studies, 2D ((1)H-(1)H) ROESY NMR spectroscopy, and electrochemical/chemical techniques. The rate law for exchange is overall second order, first order in gold(i)-thiolate and disulfide. 2D NMR experiments show evidence of association between gold(i)-thiolate and disulfide. Electrochemical/chemical investigations do not show evidence of free thiolate and are consistent with a mechanism involving formation of a [Au-S, S-S], four-centered metallacycle intermediate during gold(i)-thiolate, disulfide exchange. PMID:27353236

  19. Thiol-disulfide exchange in peptides derived from human growth hormone.

    PubMed

    Chandrasekhar, Saradha; Epling, Daniel E; Sophocleous, Andreas M; Topp, Elizabeth M

    2014-04-01

    Disulfide bonds stabilize proteins by cross-linking distant regions into a compact three-dimensional structure. They can also participate in hydrolytic and oxidative pathways to form nonnative disulfide bonds and other reactive species. Such covalent modifications can contribute to protein aggregation. Here, we present experimental data for the mechanism of thiol-disulfide exchange in tryptic peptides derived from human growth hormone in aqueous solution. Reaction kinetics was monitored to investigate the effect of pH (6.0-10.0), temperature (4-50°C), oxidation suppressants [ethylenediaminetetraacetic acid (EDTA) and N2 sparging], and peptide secondary structure (amide cyclized vs. open form). The concentrations of free thiol containing peptides, scrambled disulfides, and native disulfide-linked peptides generated via thiol-disulfide exchange and oxidation reactions were determined using reverse-phase HPLC and liquid chromatography-mass spectrometry. Concentration versus time data were fitted to a mathematical model using nonlinear least squares regression analysis. At all pH values, the model was able to fit the data with R(2) ≥ 0.95. Excluding oxidation suppressants (EDTA and N2 sparging) resulted in an increase in the formation of scrambled disulfides via oxidative pathways but did not influence the intrinsic rate of thiol-disulfide exchange. In addition, peptide secondary structure was found to influence the rate of thiol-disulfide exchange. PMID:24549831

  20. Imaging disulfide dinitroxides at 250 MHz to monitor thiol redox status

    NASA Astrophysics Data System (ADS)

    Elajaili, Hanan; Biller, Joshua R.; Rosen, Gerald M.; Kao, Joseph P. Y.; Tseytlin, Mark; Buchanan, Laura A.; Rinard, George A.; Quine, Richard W.; McPeak, Joseph; Shi, Yilin; Eaton, Sandra S.; Eaton, Gareth R.

    2015-11-01

    Measurement of thiol-disulfide redox status is crucial for characterization of tumor physiology. The electron paramagnetic resonance (EPR) spectra of disulfide-linked dinitroxides are readily distinguished from those of the corresponding monoradicals that are formed by cleavage of the disulfide linkage by free thiols. EPR spectra can thus be used to monitor the rate of cleavage and the thiol redox status. EPR spectra of 1H,14N- and 2H,15N-disulfide dinitroxides and the corresponding monoradicals resulting from cleavage by glutathione have been characterized at 250 MHz, 1.04 GHz, and 9 GHz and imaged by rapid-scan EPR at 250 MHz.

  1. Biologically oriented organic sulfur chemistry. 15. Organic disulfides and related substances. 41. Inhibition of the fungal pathogen Histoplasma capsulatum by some organic disulfides.

    PubMed

    Field, L; Grimaldi, J A; Hanley, W S; Holladay, M W; Ravichandran, R; Schaad, L J; Tate, C E

    1977-08-01

    In an extension of promising inhibitory results in vitro against Histoplasma capsulatum, correlated earlier using substituent constants developed by regression analysis with 77 disulfides, one symmetrical and 14 unsymmetrical disulfides were prepared (3--17). About half were active in vitro against H. capsulatum (and one against Candida albicans). Groups that seemed most to lead to promising inhibition among the unsymmetrical disulfides were o-HO2CC6H4, (CH2)4SO2Na, Me2NC(S), p-ClC6H4, and perhaps p-CH3C6H4; the first two also might be used to increase solubility. Earlier inhibitory promise of the morpholino group did not materialize. None of the group 3--17 was significantly active in vivo. The unsymmetrical disulfides were prepared by reaction of thiols with sulfenyl chlorides or with acyclic or cyclic thiosulfonates. Two six-membered heterocyclic disulfides (5 and 6) were prepared by a novel cyclization, in which carbon disulfide reacted with an (N-alkylamino)ethyl Bunte salt, followed by ring closure; an explanation is suggested for formation of a thiazoline when the N-alkyl group is absent. One of the disulfides disproportionated with astonishing ease (31; 0.3--1 h at 25 degrees C). PMID:330858

  2. Assisting the Assistant Principal

    ERIC Educational Resources Information Center

    Davis, James

    2008-01-01

    Retaining quality staff members is a hot topic in the public school arena. Although teachers are often the focus of concern, hiring and retaining quality assistant principals must be addressed as well. Interviewing and hiring the right assistant principal--and then ensuring that he or she remains on in a campus for several years--can do a great…

  3. Diaryl Disulfides as Novel Stabilizers of Tumor Suppressor Pdcd4

    PubMed Central

    Schmid, Tobias; Blees, Johanna S.; Bajer, Magdalena M.; Wild, Janine; Pescatori, Luca; Cuzzucoli Crucitti, Giuliana; Scipione, Luigi; Costi, Roberta; Henrich, Curtis J.; Brüne, Bernhard; Colburn, Nancy H.; Di Santo, Roberto

    2016-01-01

    The translation inhibitor and tumor suppressor Pdcd4 was reported to be lost in various tumors and put forward as prognostic marker in tumorigenesis. Decreased Pdcd4 protein stability due to PI3K-mTOR-p70S6K1 dependent phosphorylation of Pdcd4 followed by β-TrCP1-mediated ubiquitination, and proteasomal destruction of the protein was characterized as a major mechanism contributing to the loss of Pdcd4 expression in tumors. In an attempt to identify stabilizers of Pdcd4, we used a luciferase-based high-throughput compatible cellular assay to monitor phosphorylation-dependent proteasomal degradation of Pdcd4 in response to mitogen stimulation. Following a screen of approximately 2000 compounds, we identified 1,2-bis(4-chlorophenyl)disulfide as a novel Pdcd4 stabilizer. To determine an initial structure-activity relationship, we used 3 additional compounds, synthesized according to previous reports, and 2 commercially available compounds for further testing, in which either the linker between the aryls was modified (compounds 2–4) or the chlorine residues were replaced by groups with different electronic properties (compounds 5 and 6). We observed that those compounds with alterations in the sulfide linker completely lost the Pdcd4 stabilizing potential. In contrast, modifications in the chlorine residues showed only minor effects on the Pdcd4 stabilizing activity. A reporter with a mutated phospho-degron verified the specificity of the compounds for stabilizing the Pdcd4 reporter. Interestingly, the active diaryl disulfides inhibited proliferation and viability at concentrations where they stabilized Pdcd4, suggesting that Pdcd4 stabilization might contribute to the anti-proliferative properties. Finally, computational modelling indicated that the flexibility of the disulfide linker might be necessary to exert the biological functions of the compounds, as the inactive compound appeared to be energetically more restricted. PMID:26982744

  4. The ESA Geohazard Exploitation Platform

    NASA Astrophysics Data System (ADS)

    Bally, Philippe; Laur, Henri; Mathieu, Pierre-Philippe; Pinto, Salvatore

    2015-04-01

    Earthquakes represent one of the world's most significant hazards in terms both of loss of life and damages. In the first decade of the 21st century, earthquakes accounted for 60 percent of fatalities from natural disasters, according to the United Nations International Strategy for Disaster Reduction (UNISDR). To support mitigation activities designed to assess and reduce risks and improve response in emergency situations, satellite EO can be used to provide a broad range of geo-information services. This includes for instance crustal block boundary mapping to better characterize active faults, strain rate mapping to assess how rapidly faults are deforming, soil vulnerability mapping to help estimate how the soil is behaving in reaction to seismic phenomena, geo-information to assess the extent and intensity of the earthquake impact on man-made structures and formulate assumptions on the evolution of the seismic sequence, i.e. where local aftershocks or future main shocks (on nearby faults) are most likely to occur. In May 2012, the European Space Agency and the GEO Secretariat convened the International Forum on Satellite EO for Geohazards now known as the Santorini Conference. The event was the continuation of a series of international workshops such as those organized by the Geohazards Theme of the Integrated Global Observing Strategy Partnership. In Santorini the seismic community has set out a vision of the EO contribution to an operational global seismic risk program, which lead to the Geohazard Supersites and Natural Laboratories (GSNL) initiative. The initial contribution of ESA to suuport the GSNL was the first Supersites Exploitation Platform (SSEP) system in the framework of Grid Processing On Demand (GPOD), now followed by the Geohazard Exploitation Platform (GEP). In this presentation, we will describe the contribution of the GEP for exploiting satellite EO for geohazard risk assessment. It is supporting the GEO Supersites and has been further

  5. A 65 Ah rechargeable lithium molybdenum disulfide battery

    NASA Technical Reports Server (NTRS)

    Brandt, K.

    1986-01-01

    A rechargeable lithium molybdenum disulfide battery which has a number of superior performance characteristics which includes a high energy density, a high power density, and a long charge retention time was developed. The first cell sizes developed included a C size cell and an AA size cell. Over the last two years, a project to demonstrate the feasibility of the scale up to this technology to a BC size cell with 65 Ah capacity was undertaken. The objective was to develop, build, and test a .6 kWh storage battery consisting of 6 BC cells in series.

  6. Geosynchronous Performance of a Lithium-titanium Disulfide Battery

    NASA Technical Reports Server (NTRS)

    Otzinger, B.

    1985-01-01

    An ambient temperature rechargeable Lithium-Titanium disulfide (Li-TiS2) five cell battery has completed the first orbital year of accelerated synchronous orbit testing. A novel charge/discharge, state of charge (SOC) control scheme is utilized, together with taper current charge backup to overcome deleterious effects associated with high end of charge and low end of discharge voltages. It is indicated that 10 orbital years of simulated synchronous operation may be achieved. Preliminary findings associated with cell matching and battery performance are identified.

  7. Ultrafast Optical Microscopy of Single Monolayer Molybdenum Disulfide Flakes

    PubMed Central

    Seo, Minah; Yamaguchi, Hisato; Mohite, Aditya D.; Boubanga-Tombet, Stephane; Blancon, Jean-Christophe; Najmaei, Sina; Ajayan, Pulickel M.; Lou, Jun; Taylor, Antoinette J.; Prasankumar, Rohit P.

    2016-01-01

    We have performed ultrafast optical microscopy on single flakes of atomically thin CVD-grown molybdenum disulfide, using non-degenerate femtosecond pump-probe spectroscopy to excite and probe carriers above and below the indirect and direct band gaps. These measurements reveal the influence of layer thickness on carrier dynamics when probing near the band gap. Furthermore, fluence-dependent measurements indicate that carrier relaxation is primarily influenced by surface-related defect and trap states after above-bandgap photoexcitation. The ability to probe femtosecond carrier dynamics in individual flakes can thus give much insight into light-matter interactions in these two-dimensional nanosystems. PMID:26876194

  8. Ultrafast Optical Microscopy of Single Monolayer Molybdenum Disulfide Flakes

    DOE PAGESBeta

    Seo, Minah; Yamaguchi, Hisato; Mohite, Aditya D.; Boubanga-Tombet, Stephane; Blancon, Jean-Christophe; Najmaei, Sina; Ajayan, Pulickel M.; Lou, Jun; Taylor, Antoinette J.; Prasankumar, Rohit P.

    2016-02-15

    We performed ultrafast optical microscopy on single flakes of atomically thin CVD-grown molybdenum disulfide, using non-degenerate femtosecond pump-probe spectroscopy to excite and probe carriers above and below the indirect and direct band gaps. These measurements reveal the influence of layer thickness on carrier dynamics when probing near the band gap. Furthermore, fluence-dependent measurements indicate that carrier relaxation is primarily influenced by surface-related defect and trap states after above-bandgap photoexcitation. Furthermore, the ability to probe femtosecond carrier dynamics in individual flakes can thus give much insight into light-matter interactions in these two-dimensional nanosystems.

  9. First total synthesis of mycothiol and mycothiol disulfide.

    PubMed

    Lee, Sungwon; Rosazza, John P N

    2004-02-01

    [reaction: see text] The first total synthesis of mycothiol and mycothiol disulfide was achieved by linking D-2,3,4,5,6-penta-O-acetyl-myo-inositol, O-(3,4,6-tri-O-acetyl)-2-azido-2-deoxy-alpha,beta-D-glucopyranosyl) trichloroacetimidate, and N,S-diacetyl-L-cysteine and deprotecting peracetylated mycothiol. The first full spectral characterization is reported for underivatized mycothiol. The structure of mycothiol was confirmed by spectral analysis of the known bimane derivative. PMID:14748594

  10. Ultrafast Optical Microscopy of Single Monolayer Molybdenum Disulfide Flakes.

    PubMed

    Seo, Minah; Yamaguchi, Hisato; Mohite, Aditya D; Boubanga-Tombet, Stephane; Blancon, Jean-Christophe; Najmaei, Sina; Ajayan, Pulickel M; Lou, Jun; Taylor, Antoinette J; Prasankumar, Rohit P

    2016-01-01

    We have performed ultrafast optical microscopy on single flakes of atomically thin CVD-grown molybdenum disulfide, using non-degenerate femtosecond pump-probe spectroscopy to excite and probe carriers above and below the indirect and direct band gaps. These measurements reveal the influence of layer thickness on carrier dynamics when probing near the band gap. Furthermore, fluence-dependent measurements indicate that carrier relaxation is primarily influenced by surface-related defect and trap states after above-bandgap photoexcitation. The ability to probe femtosecond carrier dynamics in individual flakes can thus give much insight into light-matter interactions in these two-dimensional nanosystems. PMID:26876194

  11. Knowledge based SAR images exploitations

    NASA Astrophysics Data System (ADS)

    Wang, David L.

    1987-01-01

    One of the basic functions of SAR images exploitation system is the detection of man-made objects. The performance of object detection is strongly limited by performance of segmentation modules. This paper presents a detection paradigm composed of an adaptive segmentation algorithm based on a priori knowledge of objects followed by a top-down hierarchical detection process that generates and evaluates object hypotheses. Shadow information and inter-object relationships can be added to the knowledge base to improve performance over that of a statistical detector based only on the attributes of individual objects.

  12. Routing Algorithm Exploits Spatial Relations

    NASA Technical Reports Server (NTRS)

    Okino, Clayton; Jennings, Esther

    2004-01-01

    A recently developed routing algorithm for broadcasting in an ad hoc wireless communication network takes account of, and exploits, the spatial relationships among the locations of nodes, in addition to transmission power levels and distances between the nodes. In contrast, most prior algorithms for discovering routes through ad hoc networks rely heavily on transmission power levels and utilize limited graph-topology techniques that do not involve consideration of the aforesaid spatial relationships. The present algorithm extracts the relevant spatial-relationship information by use of a construct denoted the relative-neighborhood graph (RNG).

  13. The effect of tensile stress on the conformational free energy landscape of disulfide bonds.

    PubMed

    Anjukandi, Padmesh; Dopieralski, Przemyslaw; Ribas-Arino, Jordi; Marx, Dominik

    2014-01-01

    Disulfide bridges are no longer considered to merely stabilize protein structure, but are increasingly recognized to play a functional role in many regulatory biomolecular processes. Recent studies have uncovered that the redox activity of native disulfides depends on their C-C-S-S dihedrals, χ2 and χ'2. Moreover, the interplay of chemical reactivity and mechanical stress of disulfide switches has been recently elucidated using force-clamp spectroscopy and computer simulation. The χ2 and χ'2 angles have been found to change from conformations that are open to nucleophilic attack to sterically hindered, so-called closed states upon exerting tensile stress. In view of the growing evidence of the importance of C-C-S-S dihedrals in tuning the reactivity of disulfides, here we present a systematic study of the conformational diversity of disulfides as a function of tensile stress. With the help of force-clamp metadynamics simulations, we show that tensile stress brings about a large stabilization of the closed conformers, thereby giving rise to drastic changes in the conformational free energy landscape of disulfides. Statistical analysis shows that native TDi, DO and interchain Ig protein disulfides prefer open conformations, whereas the intrachain disulfide bridges in Ig proteins favor closed conformations. Correlating mechanical stress with the distance between the two a-carbons of the disulfide moiety reveals that the strain of intrachain Ig protein disulfides corresponds to a mechanical activation of about 100 pN. Such mechanical activation leads to a severalfold increase of the rate of the elementary redox S(N)2 reaction step. All these findings constitute a step forward towards achieving a full understanding of functional disulfides. PMID:25286308

  14. The Effect of Tensile Stress on the Conformational Free Energy Landscape of Disulfide Bonds

    PubMed Central

    Anjukandi, Padmesh; Dopieralski, Przemyslaw; Ribas–Arino, Jordi; Marx, Dominik

    2014-01-01

    Disulfide bridges are no longer considered to merely stabilize protein structure, but are increasingly recognized to play a functional role in many regulatory biomolecular processes. Recent studies have uncovered that the redox activity of native disulfides depends on their C–C–S–S dihedrals, and . Moreover, the interplay of chemical reactivity and mechanical stress of disulfide switches has been recently elucidated using force–clamp spectroscopy and computer simulation. The and angles have been found to change from conformations that are open to nucleophilic attack to sterically hindered, so–called closed states upon exerting tensile stress. In view of the growing evidence of the importance of C–C–S–S dihedrals in tuning the reactivity of disulfides, here we present a systematic study of the conformational diversity of disulfides as a function of tensile stress. With the help of force-clamp metadynamics simulations, we show that tensile stress brings about a large stabilization of the closed conformers, thereby giving rise to drastic changes in the conformational free energy landscape of disulfides. Statistical analysis shows that native TDi, DO and interchain Ig protein disulfides prefer open conformations, whereas the intrachain disulfide bridges in Ig proteins favor closed conformations. Correlating mechanical stress with the distance between the two –carbons of the disulfide moiety reveals that the strain of intrachain Ig protein disulfides corresponds to a mechanical activation of about 100 pN. Such mechanical activation leads to a severalfold increase of the rate of the elementary redox reaction step. All these findings constitute a step forward towards achieving a full understanding of functional disulfides. PMID:25286308

  15. Restoration of structural stability and ligand binding after removal of the conserved disulfide bond in tear lipocalin

    PubMed Central

    Gasymov, Oktay K.; Abduragimov, Adil R.; Glasgow, Ben J.

    2014-01-01

    Disulfide bonds play diverse structural and functional roles in proteins. In tear lipocalin (TL), the conserved sole disulfide bond regulates stability and ligand binding. Probing protein structure often involves thiol selective labeling for which removal of the disulfide bonds may be necessary. Loss of the disulfide bond may destabilize the protein so strategies to retain the native state are needed. Several approaches were tested to regain the native conformational state in the disulfide-less protein. These included the addition of thrimethylamine N-oxide (TMAO) and the substitution of the Cys residues of disulfide bond with residues that can either form a potential salt bridge or others that can create a hydrophobic interaction. TMAO stabilized the protein relaxed by removal of the disulfide bond. In the disulfide-less mutants of TL, 1.0 M TMAO increased the free energy change (ΔG0) significantly from 2.1 to 3.8 kcal/mol. Moderate recovery was observed for the ligand binding tested with NBD-cholesterol. Because the disulfide bond of TL is solvent exposed, the substitution of the disulfide bond with a potential salt bridge or hydrophobic interaction did not stabilize the protein. This approach should work for buried disulfide bonds. However, for proteins with solvent exposed disulfide bonds, the use of TMAO may be an excellent strategy to restore the native conformational states in disulfide-less analogs of the proteins. PMID:25223802

  16. Restoration of structural stability and ligand binding after removal of the conserved disulfide bond in tear lipocalin.

    PubMed

    Gasymov, Oktay K; Abduragimov, Adil R; Glasgow, Ben J

    2014-10-01

    Disulfide bonds play diverse structural and functional roles in proteins. In tear lipocalin (TL), the conserved sole disulfide bond regulates stability and ligand binding. Probing protein structure often involves thiol selective labeling for which removal of the disulfide bonds may be necessary. Loss of the disulfide bond may destabilize the protein so strategies to retain the native state are needed. Several approaches were tested to regain the native conformational state in the disulfide-less protein. These included the addition of trimethylamine N-oxide (TMAO) and the substitution of the Cys residues of disulfide bond with residues that can either form a potential salt bridge or others that can create a hydrophobic interaction. TMAO stabilized the protein relaxed by removal of the disulfide bond. In the disulfide-less mutants of TL, 1.0M TMAO increased the free energy change (ΔG(0)) significantly from 2.1 to 3.8kcal/mol. Moderate recovery was observed for the ligand binding tested with NBD-cholesterol. Because the disulfide bond of TL is solvent exposed, the substitution of the disulfide bond with a potential salt bridge or hydrophobic interaction did not stabilize the protein. This approach should work for buried disulfide bonds. However, for proteins with solvent exposed disulfide bonds, the use of TMAO may be an excellent strategy to restore the native conformational states in disulfide-less analogs of the proteins. PMID:25223802

  17. Legionella pneumophila utilizes a Single Player Disulfide-Bond Oxidoreductase System to Manage Disulfide Bond Formation and Isomerization

    PubMed Central

    Kpadeh, Zegbeh Z.; Day, Shandra R.; Mills, Brandy W.; Hoffman, Paul S.

    2015-01-01

    Legionella pneumophila uses a single homodimeric disulfide bond (DSB) oxidoreductase DsbA2 to catalyze extracytoplasmic protein folding and to correct DSB errors through protein-disulfide isomerase (PDI) activity. In Escherichia coli, these functions are separated to avoid futile cycling. In L. pneumophila, DsbA2 is maintained as a mixture of disulfides (S-S) and free thiols (SH), but when expressed in E. coli, only the SH form is observed. We provide evidence to suggest that structural differences in DsbB oxidases (LpDsbB1 and LpDsbB2) and DsbD reductases (LpDsbD1 and LpDsbD2) (compared to E. coli) permit bifunctional activities without creating a futile cycle. LpdsbB1 and LpdsbB2 partially complemented an EcdsbB mutant while neither LpdsbD1 nor LpdsbD2 complemented an EcdsbD mutant unless DsbA2 was also expressed. When the dsb genes of E. coli were replaced with those of L. pneumophila, motility was restored and DsbA2 was present as a mixture of redox forms. A dominant-negative approach to interfere with DsbA2 function in L. pneumophila determined that DSB oxidase activity was necessary for intracellular multiplication and assembly/function of the Dot/Icm Type IVb secretion system. Our studies show that a single-player system may escape the futile cycle trap by limiting transfer of reducing equivalents from LpDsbDs to DsbA2. PMID:25534767

  18. National Center for Missing and Exploited Children

    MedlinePlus

    ... child Call 1-800-843-5678 Report sexual exploitation of a child The CyberTipline ® receives leads and tips regarding suspected crimes of sexual exploitation committed against children. More than 3.3 million ...

  19. A laboratory and theoretical study of protonated carbon disulfide, HSCS+.

    PubMed

    McCarthy, M C; Thaddeus, P; Wilke, Jeremiah J; Schaefer, Henry F

    2009-06-21

    The rotational spectrum of protonated carbon disulfide, HSCS(+), has been detected in the centimeter-wave band in a molecular beam by Fourier transform microwave spectroscopy. Rotational and centrifugal distortion constants have been determined from ten transitions in the K(a)=0 ladder of the normal isotopic species, HS(13)CS(+), and DSCS(+). The present assignment agrees well with high-level coupled cluster calculations of the HSCS(+) structure, which, like earlier work, predict this isomer to be the ground state on the HCS(2) (+) potential energy surface; HCSS(+), an isomer with C(2v) symmetry, is predicted to lie more than 20 kcal/mol higher in energy. Other properties of HSCS(+) including its dipole moment, anharmonic vibrational frequencies, and infrared intensities have also been computed at the coupled cluster level of theory with large basis sets. Because carbon disulfide possesses a fairly large proton affinity, and because this nonpolar molecule may plausibly exist in astronomical sources, HSCS(+) is a good candidate for detection with radio telescopes in the submillimeter band where the stronger b-type transitions of this protonated cation are predicted to lie. PMID:19548724

  20. Diversity of the Epsilonproteobacteria Dsb (disulfide bond) systems

    PubMed Central

    Bocian-Ostrzycka, Katarzyna M.; Grzeszczuk, Magdalena J.; Dziewit, Lukasz; Jagusztyn-Krynicka, Elżbieta K.

    2015-01-01

    The bacterial proteins of the Dsb family—important components of the post-translational protein modification system—catalyze the formation of disulfide bridges, a process that is crucial for protein structure stabilization and activity. Dsb systems play an essential role in the assembly of many virulence factors. Recent rapid advances in global analysis of bacteria have thrown light on the enormous diversity among bacterial Dsb systems. While the Escherichia coli disulfide bond-forming system is quite well understood, the mechanisms of action of Dsb systems in other bacteria, including members of class Epsilonproteobacteria that contain pathogenic and non-pathogenic bacteria colonizing extremely diverse ecological niches, are poorly characterized. Here we present a review of current knowledge on Epsilonproteobacteria Dsb systems. We have focused on the Dsb systems of Campylobacter spp. and Helicobacter spp. because our knowledge about Dsb proteins of Wolinella and Arcobacter spp. is still scarce and comes mainly from bioinformatic studies. Helicobacter pylori is a common human pathogen that colonizes the gastric epithelium of humans with severe consequences. Campylobacter spp. is a leading cause of zoonotic enteric bacterial infections in most developed and developing nations. We focus on various aspects of the diversity of the Dsb systems and their influence on pathogenicity, particularly because Dsb proteins are considered as potential targets for a new class of anti-virulence drugs to treat human infections by Campylobacter or Helicobacter spp. PMID:26106374

  1. Lithium/disulfide cells capable of long cycle life

    SciTech Connect

    Kaun, T.D.; Holifield, T.F.; DeLuca, W.H.

    1988-01-01

    The lithium-alloy/disulfide cell has undergone improvements to provide a very stable, high performance upper-plateau (UP) FeS/sub 2/ electrode. Prismatic UP FeS/sub 2/ cell tests (12--24 Ah capacity) with a LiCl-LiBr-KBr eutectic electrolyte have demonstrated 1000 deep discharge cycles at 400/degree/C with less than a 20% drop in capacity and without reduced power capability. Previous lithium-alloy/disulfide cells, which were based on a two voltage-plateau FeS/sub 2/ electrode and LiCl-KCl eutectic electrolyte had a life expectancy of only 100 cycles. Both time- and cycle-related capacity loss mechanisms have been eliminated with the improved cell design. In addition, new cell design features of overcharge tolerance and overdischarge safeguarding enhance battery durability. The performance prospects of a Li-alloy/UP FeS/sub 2/ battery for an IDSEP van application are discussed. A specific energy of 150 Wh/kg for this battery after 1000 cycles of operation is projected. 8 refs., 5 figs., 1 tab.

  2. Methods of measuring Protein Disulfide Isomerase activity: a critical overview

    NASA Astrophysics Data System (ADS)

    Watanabe, Monica; Laurindo, Francisco; Fernandes, Denise

    2014-09-01

    Protein disulfide isomerase is an essential redox chaperone from the endoplasmic reticulum (ER) and is responsible for correct disulfide bond formation in nascent proteins. PDI is also found in other cellular locations in the cell, particularly the cell surface. Overall, PDI contributes to ER and global cell redox homeostasis and signaling. The knowledge about PDI structure and function progressed substantially based on in vitro studies using recombinant PDI and chimeric proteins. In these experimental scenarios, PDI reductase and chaperone activities are readily approachable. In contrast, assays to measure PDI isomerase activity, the hallmark of PDI family, are more complex. Assessment of PDI roles in cells and tissues mainly relies on gain- or loss-of-function studies. However, there is limited information regarding correlation of experimental readouts with the distinct types of PDI activities. In this mini-review, we evaluate the main methods described for measuring the different kinds of PDI activity: thiol reductase, thiol oxidase, thiol isomerase and chaperone. We emphasize the need to use appropriate controls and the role of critical interferents (e.g., detergent, presence of reducing agents). We also discuss the translation of results from in vitro studies with purified recombinant PDI to cellular and tissue samples, with critical comments on the interpretation of results.

  3. Protected peptide disulfides by oxidative detachment from a support.

    PubMed

    Rietman, B H; Smulders, R H; Eggen, I F; van Vliet, A; van de Werken, G; Tesser, G I

    1994-09-01

    A new and efficient procedure for the preparation of protected cyclized and protected symmetrical dimeric peptide disulfides is described. A thiol is immobilized onto a solid phase through coupling of the thiol function with a resin-linked trityl group. Following conventional peptide assembly using the Fmoc-strategy, detachment is performed by oxidation with iodine in a suitable organic solvent. When N,N-dimethylformamide is used as the solvent, and the peptide chain contains an acetamidomethylthio function, located N-terminally in a N alpha-(9-fluorenylmethyloxycarbonyl), or N alpha-tert-butyloxycarbonyl cysteinyl residue, or occurring in the chain, then the corresponding fully protected cyclic peptide disulfide will be obtained in high yield and purity. In other solvents (e.g. dioxane or chloroform-methanol 1:1, v/v), the iodine-mediated oxidation gave not only the cyclic product, but also substantial amounts of the parallel symmetrical dimeric peptide retaining Cys(Acm) at the two identical N-termini. PMID:7822095

  4. Biotechnology Based Process for Production of a Disulfide-Bridged Peptide.

    PubMed

    Goswami, Animesh; Goldberg, Steven L; Hanson, Ronald L; Johnston, Robert M; Lyngberg, Olav K; Chan, Yeung; Lo, Ehrlic; Chan, Steven H; de Mas, Nuria; Ramirez, Antonio; Doyle, Richard; Ding, Wei; Gao, Mian; Krystek, Stanley R; Wan, Changhong; Kim, Yeoun Jin; Calambur, Deepa; Witmer, Mark; Bryson, James W

    2016-05-18

    A disulfide-bridged peptide drug development candidate contained two oligopeptide chains with 11 and 12 natural amino acids joined by a disulfide bond at the N-terminal end. An efficient biotechnology based process for the production of the disulfide-bridged peptide was developed. Initially, the two individual oligopeptide chains were prepared separately by designing different fusion proteins and expressing them in recombinant E. coli. Enzymatic or chemical cleavage of the two fusion proteins provided the two individual oligopeptide chains which could be conjugated via disulfide bond by conventional chemical reaction to the disulfide-bridged peptide. A novel heterodimeric system to bring the two oligopeptide chains closer and induce disulfide bond formation was designed by taking advantage of the self-assembly of a leucine zipper system. The heterodimeric approach involved designing fusion proteins with the acidic and basic components of the leucine zipper, additional amino acids to optimize interaction between the individual chains, specific cleavage sites, specific tag to ensure separation, and two individual oligopeptide chains. Computer modeling was used to identify the nature and number of amino acid residue to be inserted between the leucine zipper and oligopeptides for optimum interaction. Cloning and expression in rec E. coli, fermentation, followed by cell disruption resulted in the formation of heterodimeric protein with the interchain disulfide bond. Separation of the desired heterodimeric protein, followed by specific cleavage at methionine by cyanogen bromide provided the disulfide-bridged peptide. PMID:27098672

  5. Regulation of interleukin-4 signaling by extracellular reduction of intramolecular disulfides

    SciTech Connect

    Curbo, Sophie; Gaudin, Raphael; Carlsten, Mattias; Malmberg, Karl-Johan; Troye-Blomberg, Marita; Ahlborg, Niklas; Karlsson, Anna; Johansson, Magnus; Lundberg, Mathias

    2009-12-25

    Interleukin-4 (IL-4) contains three structurally important intramolecular disulfides that are required for the bioactivity of the cytokine. We show that the cell surface of HeLa cells and endotoxin-activated monocytes can reduce IL-4 intramolecular disulfides in the extracellular space and inhibit binding of IL-4 to the IL-4R{alpha} receptor. IL-4 disulfides were in vitro reduced by thioredoxin 1 (Trx1) and protein disulfide isomerase (PDI). Reduction of IL-4 disulfides by the cell surface of HeLa cells was inhibited by auranofin, an inhibitor of thioredoxin reductase that is an electron donor to both Trx1 and PDI. Both Trx1 and PDI have been shown to be located at the cell surface and our data suggests that these enzymes are involved in catalyzing reduction of IL-4 disulfides. The pro-drug N-acetylcysteine (NAC) that promotes T-helper type 1 responses was also shown to mediate the reduction of IL-4 disulfides. Our data provides evidence for a novel redox dependent pathway for regulation of cytokine activity by extracellular reduction of intramolecular disulfides at the cell surface by members of the thioredoxin enzyme family.

  6. 46 CFR 151.50-40 - Additional requirements for carbon disulfide (carbon bisulfide) and ethyl ether.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 5 2014-10-01 2014-10-01 false Additional requirements for carbon disulfide (carbon... Special Requirements § 151.50-40 Additional requirements for carbon disulfide (carbon bisulfide) and ethyl... waterways at the loading and unloading points. (f) The special requirements of § 151.50-41 for...

  7. 21 CFR 520.1802 - Piperazine-carbon disulfide complex oral dosage forms.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Piperazine-carbon disulfide complex oral dosage forms. 520.1802 Section 520.1802 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... § 520.1802 Piperazine-carbon disulfide complex oral dosage forms....

  8. 21 CFR 520.1802 - Piperazine-carbon disulfide complex oral dosage forms.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Piperazine-carbon disulfide complex oral dosage forms. 520.1802 Section 520.1802 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... § 520.1802 Piperazine-carbon disulfide complex oral dosage forms....

  9. 21 CFR 520.1802 - Piperazine-carbon disulfide complex oral dosage forms.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Piperazine-carbon disulfide complex oral dosage forms. 520.1802 Section 520.1802 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... § 520.1802 Piperazine-carbon disulfide complex oral dosage forms....

  10. 46 CFR 151.50-40 - Additional requirements for carbon disulfide (carbon bisulfide) and ethyl ether.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 5 2012-10-01 2012-10-01 false Additional requirements for carbon disulfide (carbon... Special Requirements § 151.50-40 Additional requirements for carbon disulfide (carbon bisulfide) and ethyl... waterways at the loading and unloading points. (f) The special requirements of § 151.50-41 for...

  11. 21 CFR 520.1802 - Piperazine-carbon disulfide complex oral dosage forms.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Piperazine-carbon disulfide complex oral dosage forms. 520.1802 Section 520.1802 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... § 520.1802 Piperazine-carbon disulfide complex oral dosage forms....

  12. 46 CFR 151.50-40 - Additional requirements for carbon disulfide (carbon bisulfide) and ethyl ether.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Additional requirements for carbon disulfide (carbon... Special Requirements § 151.50-40 Additional requirements for carbon disulfide (carbon bisulfide) and ethyl... waterways at the loading and unloading points. (f) The special requirements of § 151.50-41 for...

  13. 21 CFR 520.1802 - Piperazine-carbon disulfide complex oral dosage forms.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Piperazine-carbon disulfide complex oral dosage forms. 520.1802 Section 520.1802 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... § 520.1802 Piperazine-carbon disulfide complex oral dosage forms....

  14. 46 CFR 151.50-40 - Additional requirements for carbon disulfide (carbon bisulfide) and ethyl ether.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 5 2013-10-01 2013-10-01 false Additional requirements for carbon disulfide (carbon... Special Requirements § 151.50-40 Additional requirements for carbon disulfide (carbon bisulfide) and ethyl... waterways at the loading and unloading points. (f) The special requirements of § 151.50-41 for...

  15. 46 CFR 151.50-40 - Additional requirements for carbon disulfide (carbon bisulfide) and ethyl ether.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 5 2011-10-01 2011-10-01 false Additional requirements for carbon disulfide (carbon... Special Requirements § 151.50-40 Additional requirements for carbon disulfide (carbon bisulfide) and ethyl... waterways at the loading and unloading points. (f) The special requirements of § 151.50-41 for...

  16. Selective inhibition of nicotinamide adenine dinucleotide kinases by dinucleoside disulfide mimics of nicotinamide adenine dinucleotide analogues.

    PubMed

    Petrelli, Riccardo; Sham, Yuk Yin; Chen, Liqiang; Felczak, Krzysztof; Bennett, Eric; Wilson, Daniel; Aldrich, Courtney; Yu, Jose S; Cappellacci, Loredana; Franchetti, Palmarisa; Grifantini, Mario; Mazzola, Francesca; Di Stefano, Michele; Magni, Giulio; Pankiewicz, Krzysztof W

    2009-08-01

    Diadenosine disulfide (5) was reported to inhibit NAD kinase from Listeria monocytogenes and the crystal structure of the enzyme-inhibitor complex has been solved. We have synthesized tiazofurin adenosine disulfide (4) and the disulfide 5, and found that these compounds were moderate inhibitors of human NAD kinase (IC(50)=110 microM and IC(50)=87 microM, respectively) and Mycobacterium tuberculosis NAD kinase (IC(50)=80 microM and IC(50)=45 microM, respectively). We also found that NAD mimics with a short disulfide (-S-S-) moiety were able to bind in the folded (compact) conformation but not in the common extended conformation, which requires the presence of a longer pyrophosphate (-O-P-O-P-O-) linkage. Since majority of NAD-dependent enzymes bind NAD in the extended conformation, selective inhibition of NAD kinases by disulfide analogues has been observed. Introduction of bromine at the C8 of the adenine ring restricted the adenosine moiety of diadenosine disulfides to the syn conformation making it even more compact. The 8-bromoadenosine adenosine disulfide (14) and its di(8-bromoadenosine) analogue (15) were found to be the most potent inhibitors of human (IC(50)=6 microM) and mycobacterium NAD kinase (IC(50)=14-19 microM reported so far. None of the disulfide analogues showed inhibition of lactate-, and inosine monophosphate-dehydrogenase (IMPDH), enzymes that bind NAD in the extended conformation. PMID:19596199

  17. The exploitation argument against commercial surrogacy.

    PubMed

    Wilkinson, Stephen

    2003-04-01

    This paper discusses the exploitation argument against commercial surrogacy: the claim that commercial surrogacy is morally objectionable because it is exploitative. The following questions are addressed. First, what exactly does the exploitation argument amount to? Second, is commercial surrogacy in fact exploitative? Third, if it were exploitative, would this provide a sufficient reason to prohibit (or otherwise legislatively discourage) it? The focus throughout is on the exploitation of paid surrogates, although it is noted that other parties (e.g. 'commissioning parents') may also be the victims of exploitation. It is argued that there are good reasons for believing that commercial surrogacy is often exploitative. However, even if we accept this, the exploitation argument for prohibiting (or otherwise legislatively discouraging) commercial surrogacy remains quite weak. One reason for this is that prohibition may well 'backfire' and lead to potential surrogates having to do other things that are more exploitative and/or more harmful than paid surrogacy. It is concluded therefore that those who oppose exploitation should (rather than attempting to stop particular practices like commercial surrogacy) concentrate on: (a) improving the conditions under which paid surrogates 'work'; and (b) changing the background conditions (in particular, the unequal distribution of power and wealth) which generate exploitative relationships. PMID:12812183

  18. Selective Dynamic Assembly of Disulfide Macrocyclic Helical Foldamers with Remote Communication of Handedness.

    PubMed

    Tsiamantas, Christos; de Hatten, Xavier; Douat, Céline; Kauffmann, Brice; Maurizot, Victor; Ihara, Hirotaka; Takafuji, Makoto; Metzler-Nolte, Nils; Huc, Ivan

    2016-06-01

    Disulfide bridge formation was investigated in helical aromatic oligoamide foldamers. Depending on the position of thiol-bearing side chains, exclusive intramolecular or intermolecular disulfide bridging may occur. The two processes are capable of self-sorting, presumably by dynamic exchange. Quantitative assessment of helix handedness inversion rates showed that bridging stabilizes the folded structures. Intermolecular disulfide bridging serendipitously yielded a well-defined, C2 -symmetrical, two-helix bundle-like macrocyclic structure in which complete control over relative handedness, that is, helix-helix handedness communication, is mediated remotely by the disulfide bridged side chains in the absence of contacts between helices. MM calculations suggest that this phenomenon is specific to a given side chain length and requires disulfide functions. PMID:27101565

  19. Energy for lunar resource exploitation

    NASA Technical Reports Server (NTRS)

    Glaser, Peter E.

    1992-01-01

    Humanity stands at the threshold of exploiting the known lunar resources that have opened up with the access to space. America's role in the future exploitation of space, and specifically of lunar resources, may well determine the level of achievement in technology development and global economic competition. Space activities during the coming decades will significantly influence the events on Earth. The 'shifting of history's tectonic plates' is a process that will be hastened by the increasingly insistent demands for higher living standards of the exponentially growing global population. Key to the achievement of a peaceful world in the 21st century, will be the development of a mix of energy resources at a societally acceptable and affordable cost within a realistic planning horizon. This must be the theme for the globally applicable energy sources that are compatible with the Earth's ecology. It is in this context that lunar resources development should be a primary goal for science missions to the Moon, and for establishing an expanding human presence. The economic viability and commercial business potential of mining, extracting, manufacturing, and transporting lunar resource based materials to Earth, Earth orbits, and to undertake macroengineering projects on the Moon remains to be demonstrated. These extensive activities will be supportive of the realization of the potential of space energy sources for use on Earth. These may include generating electricity for use on Earth based on beaming power from Earth orbits and from the Moon to the Earth, and for the production of helium 3 as a fuel for advanced fusion reactors.

  20. Conformational analysis and design of cross-strand disulfides in antiparallel β-sheets.

    PubMed

    Indu, S; Kochat, V; Thakurela, S; Ramakrishnan, C; Varadarajan, Raghavan

    2011-01-01

    Cross-strand disulfides bridge two cysteines in a registered pair of antiparallel β-strands. A nonredundant data set comprising 5025 polypeptides containing 2311 disulfides was used to study cross-strand disulfides. Seventy-six cross-strand disulfides were found of which 75 and 1 occurred at non-hydrogen-bonded (NHB) and hydrogen-bonded (HB) registered pairs, respectively. Conformational analysis and modeling studies demonstrated that disulfide formation at HB pairs necessarily requires an extremely rare and positive χ¹ value for at least one of the cysteine residues. Disulfides at HB positions also have more unfavorable steric repulsion with the main chain. Thirteen pairs of disulfides were introduced in NHB and HB pairs in four model proteins: leucine binding protein (LBP), leucine, isoleucine, valine binding protein (LIVBP), maltose binding protein (MBP), and Top7. All mutants LIVBP T247C V331C showed disulfide formation either on purification, or on treatment with oxidants. Protein stability in both oxidized and reduced states of all mutants was measured. Relative to wild type, LBP and MBP mutants were destabilized with respect to chemical denaturation, although the sole exposed NHB LBP mutant showed an increase of 3.1°C in T(m). All Top7 mutants were characterized for stability through guanidinium thiocyanate chemical denaturation. Both exposed and two of the three buried NHB mutants were appreciably stabilized. All four HB Top7 mutants were destabilized (ΔΔG⁰ = -3.3 to -6.7 kcal/mol). The data demonstrate that introduction of cross-strand disulfides at exposed NHB pairs is a robust method of improving protein stability. All four exposed Top7 disulfide mutants showed mild redox activity. PMID:21058397

  1. The Disulfide Bonds within BST-2 Enhance Tensile Strength during Viral Tethering.

    PubMed

    Du Pont, Kelly E; McKenzie, Aidan M; Kokhan, Oleksandr; Sumner, Isaiah; Berndsen, Christopher E

    2016-02-16

    Human BST-2/tetherin is a host factor that inhibits the release of enveloped viruses, including HIV-1, HIV-2, and SIV, from the cell surface by tethering viruses to the host cell membrane. BST-2 has an α-helical ectodomain that forms disulfide-linked dimers between two monomers forming a coiled coil. The ectodomain contains three cysteine residues that can participate in disulfide bond formation and are critical for viral tethering. The role of the disulfides in viral tethering is unknown but proposed to be for maintaining the dimer. We explored the role of the disulfides in the structure of BST-2 using experimental, biophysical methods. To understand the role of the disulfides in viral tethering, we used a new approach in viral tethering, steered molecular dynamics. We find that the disulfides coordinate the unfolding of the BST-2 monomers, which adds tensile strength to the coiled coil. Structural differences between oxidized and reduced BST-2 are apparent during unfolding, showing the monomers slide past each other in the absence of the disulfides. We found no evidence to support dissociation of the dimer upon reduction of the disulfide bonds. Moreover, the structure of BST-2 in the absence of the disulfides is similar to that of the oxidized form of BST-2, supporting previous X-ray crystallography and cellular work that showed the disulfides are not required for expression of BST-2. These data provide new insights into viral tethering by using novel techniques in the analysis of BST-2 to give amino acid level insight into functions of BST-2. PMID:26789136

  2. Cell-free synthesis system suitable for disulfide-containing proteins

    SciTech Connect

    Matsuda, Takayoshi; Watanabe, Satoru; Kigawa, Takanori

    2013-02-08

    Highlights: ► Cell-free synthesis system suitable for disulfide-containing proteins is proposed. ► Disulfide bond formation was facilitated by the use of glutathione buffer. ► DsbC catalyzed the efficient shuffling of incorrectly formed disulfide bonds. ► Milligram quantities of functional {sup 15}N-labeled BPTI and lysozyme C were obtained. ► Synthesized proteins were both catalytically functional and properly folded. -- Abstract: Many important therapeutic targets are secreted proteins with multiple disulfide bonds, such as antibodies, cytokines, hormones, and proteases. The preparation of these proteins for structural and functional analyses using cell-based expression systems still suffers from several issues, such as inefficiency, low yield, and difficulty in stable-isotope labeling. The cell-free (or in vitro) protein synthesis system has become a useful protein production method. The openness of the cell-free system allows direct control of the reaction environment to promote protein folding, making it well suited for the synthesis of disulfide-containing proteins. In this study, we developed the Escherichia coli (E. coli) cell lysate-based cell-free synthesis system for disulfide-containing proteins, which can produce sufficient amounts of functional proteins for NMR analyses. Disulfide bond formation was facilitated by the use of glutathione buffer. In addition, disulfide isomerase, DsbC, catalyzed the efficient shuffling of incorrectly formed disulfide bonds during the protein synthesis reaction. We successfully synthesized milligram quantities of functional {sup 15}N-labeled higher eukaryotic proteins, bovine pancreatic trypsin inhibitor (BPTI) and human lysozyme C (LYZ). The NMR spectra and functional analyses indicated that the synthesized proteins are both catalytically functional and properly folded. Thus, the cell-free system is useful for the synthesis of disulfide-containing proteins for structural and functional analyses.

  3. Disulfide structures of highly bridged peptides: a new strategy for analysis.

    PubMed Central

    Gray, W. R.

    1993-01-01

    A new approach is described for analyzing disulfide linkage patterns in peptides containing tightly clustered cystines. Such peptides are very difficult to analyze with traditional strategies, which require that the peptide chain be split between close or adjacent Cys residues. The water-soluble tris-(2-carboxyethyl)-phosphine (TCEP) reduced disulfides at pH 3, and partially reduced peptides were purified by high performance liquid chromatography with minimal thiol-disulfide exchange. Alkylation of free thiols, followed by sequencer analysis, provided explicit assignment of disulfides that had been reduced. Thiol-disulfide exchange occurred during alkylation of some peptides, but correct deductions were still possible. Alkylation competed best with exchange when peptide solution was added with rapid mixing to 2.2 M iodoacetamide. Variants were developed in which up to three alkylating agents were used to label different pairs of thiols, allowing a full assignment in one sequencer analysis. Model peptides used included insulin (three bridges, intra- and interchain disulfides; -Cys.Cys- pair), endothelin and apamin (two disulfides; -Cys.x.Cys- pair), conotoxin GI and isomers (two disulfides; -Cys.Cys- pair), and bacterial enterotoxin (three bridges within 13 residues; two -Cys.Cys- pairs). With insulin, all intermediates in the reduction pathway were identified; with conotoxin GI, analysis was carried out successfully for all three disulfide isomers. In addition to these known structures, the method has been applied successfully to the analysis of several previously unsolved structures of similar complexity. Rates of reduction of disulfide bonds varied widely, but most peptides did not show a strongly preferred route for reduction. PMID:8251945

  4. The role of thermal excitation in the tunneling-electron-induced reaction: Dissociation of dimethyl disulfide on Cu(111)

    NASA Astrophysics Data System (ADS)

    Motobayashi, Kenta; Kim, Yousoo; Ohara, Michiaki; Ueba, Hiromu; Kawai, Maki

    2016-01-01

    We found a thermally assisted increase in anharmonic coupling between the reaction coordinate and Csbnd H(D) stretch mode for the dissociation of a single dimethyl disulfide molecule on Cu(111) induced by inelastic tunneling electrons from a tip of scanning tunneling microscope (STM). The reaction order, i.e. the number of electrons required for a reaction, changes from two to one at elevated temperature while the Csbnd H(D) stretch mode is excited by tunneling electrons. The detailed reaction mechanism is studied through the quantitative analysis of the non-integer reaction order observed at intermediate temperature, where low energy vibrational mode originated from the hybridized state of molecule and substrate plays a key role.

  5. Characterization of IgG2 Disulfide Bonds with LC/MS/MS and Postcolumn Online Reduction.

    PubMed

    Liu, Hongji; Lei, Qing Paula; Washabaugh, Michael

    2016-05-17

    The complication of IgG2 disulfide connections demands advances in techniques for disulfide bond determination. We have developed a new LC/MS/MS method for improved disulfide analysis. With postcolumn introduction of dithiothreitol (DTT) and ammonium hydroxide, each disulfide-containing peptide eluted out of LC in an acidic mobile phase can be rapidly reduced prior to MS analysis. The reduction can be driven to near completion. The reagents are MS-friendly, and the reaction occurs at no cost of separation (little is added to the postcolumn dead volume of the LC system). Comparing LC/MS data with and without online reduction, a direct correlation can be established between a disulfide peptide and its composing peptides using retention time. With disulfide online removal, high-quality MS/MS fragmentation data can be acquired and allows for definitive determination of the disulfide peptide. This technique is especially valuable in determining the disulfide bond linkage of complicated molecules such as the hinge-containing disulfide peptides produced from IgG2 disulfide isoforms. Due to over/under enzymatic cleavages, multiple hinge-containing disulfide peptides are produced from each isoform. Twenty-two hinge-containing disulfide peptides in total have been confidently identified with this technique. Without the method, successful identification to many of these peptides would have become extremely difficult. PMID:27111505

  6. Evaluation of disulfide scrambling during the enzymatic digestion of bevacizumab at various pH values using mass spectrometry.

    PubMed

    Sung, Wang-Chou; Chang, Chiung-Wen; Huang, Sheng-Yu; Wei, Ting-Yu; Huang, Yi-Li; Lin, Yu-Hua; Chen, Han-Min; Chen, Sung-Fang

    2016-09-01

    Disulfide linkages play an important role in protein stability and activity. Thus, it is critical to characterize disulfide bonds to ensure the quality and function of protein pharmaceuticals. There are, however, problems associated with maintaining disulfide linkages in the conventional procedures that are used to digest a protein. In order to preserve enzyme activity during the digestion of a protein, it is commonly carried out at neutral to basic environment which increases the possibilities of disulfide bond scrambling. However, it is not easy to differentiate whether the scrambled disulfide linkages are initiated by the sample itself or whether they are induced during the protease digestion process. In this study, the optimum pH for minimizing disulfide bond rearrangements during the digestion process was determined. Three sets of proteases, trypsin plus Glu-C, Lys-C and thermolysin were used, followed by dimethyl labeling and mass spectrometry for a bevacizumab (Avastin) disulfide linkage analysis. No disulfide linkage scrambling was detected at pH6 when Lys-C or trypsin plus Glu-C were used as enzymes. When thermolysin was applied, some scrambled disulfide bonds were identified at pH5, 6 and 7. Nevertheless, there was less disulfide bond scrambling at a lower pH. All correct disulfide bonds on bevacizumab could be identified using this approach. The results demonstrated that by choosing the proper enzymes, using a lower pH environment for the digestion could reduce the degree of artifact disulfide scrambling. PMID:27238563

  7. Opportunistic exploitation: an overlooked pathway to extinction.

    PubMed

    Branch, Trevor A; Lobo, Aaron S; Purcell, Steven W

    2013-07-01

    How can species be exploited economically to extinction? Past single-species hypotheses examining the economic plausibility of exploiting rare species have argued that the escalating value of rarity allows extinction to be profitable. We describe an alternative pathway toward extinction in multispecies exploitation systems, termed 'opportunistic exploitation'. In this mode, highly valued species that are targeted first by fishing, hunting, and logging become rare, but their populations can decline further through opportunistic exploitation while more common but less desirable species are targeted. Effectively, expanding exploitation to more species subsidizes the eventual extinction of valuable species at low densities. Managers need to recognize conditions that permit opportunistic depletion and pass regulations to protect highly desirable species when exploitation can expand to other species. PMID:23562732

  8. A conserved disulfide motif in human tear lipocalins influences ligand binding.

    PubMed

    Glasgow, B J; Abduragimov, A R; Yusifov, T N; Gasymov, O K; Horwitz, J; Hubbell, W L; Faull, K F

    1998-02-24

    Structural and functional characteristics of the disulfide motif have been determined for tear lipocalins, members of a novel group of proteins that carry lipids. Amino acid sequences for two of the six isolated isoforms were assigned by a comparison of molecular mass measurements with masses calculated from the cDNA-predicted protein sequence and available N-terminal protein sequence data. A third isoform was tentatively sequence assigned using the same criteria. The most abundant isoform has a measured mass of 17 446.3 Da, consistent with residues 19-176 of the putative precursor (calculated mass 17 445.8 Da). Chemical derivatization of native and reduced/denatured protein confirmed the presence of a single intramolecular disulfide bond in the native protein. Reactivity of native, reduced, and denatured protein with 4-pyridine disulfide and dithiobis(2-nitrobenzoic acid) indicated that access to the free cysteine is markedly restricted by the intact disulfide bridge. Mass measurements of tryptic fragments identified C119 as the free cysteine and showed that the single intramolecular disulfide bond joined residues C79 and C171. Circular dichroism indicated that tear lipocalins have a predominant beta-pleated sheet structure (44%) that is essentially retained after reduction of the disulfide bond. Circular dichroism in the far-UV showed reduced molecular asymmetry and enhanced urea-induced unfolding with disulfide reduction indicative of relaxation of protein structure. Circular dichroism in the near-UV shows that the disulfide bond contributes to the asymmetry of aromatic sites. The effect of disulfide reduction on ligand binding was monitored using the intrinsic optical activity of bound retinol. The intact disulfide bond diminishes the affinity of tear lipocalins for retinol and restricts the displacement of native lipids by retinol. Disulfide reduction is accompanied by a dramatic alteration in ligand-induced conformational changes that involves aromatic

  9. Temperature effect on optical spectra of monolayer molybdenum disulfide

    SciTech Connect

    Soklaski, Ryan; Liang, Yufeng; Yang, Li

    2014-05-12

    Recently, measured optical absorption and photoluminescence spectra reveal substantial frequency shifts of both exciton and trion peaks as monolayer molybdenum disulfide, MoS{sub 2}, is cooled from 363 K to 4 K. First-principles simulations using the GW-Bethe-Salpeter equation approach satisfactorily reproduce these frequency shifts by incorporating the thermal expansion effect. Studying these temperature effects in monolayer MoS{sub 2} is crucial for rectifying the results of available experiments with the previous predictions of zero-temperature-limit simulations. Moreover, our estimated thermal expansion coefficient of monolayer MoS{sub 2} is less than that of bulk counterpart by tracking the frequency shifts of the exciton peaks in optical spectra. This may serve as a convenient way to estimate thermal expansion coefficients of general two-dimensional chalcogenides.

  10. High Performance Molybdenum Disulfide Amorphous Silicon Heterojunction Photodetector

    PubMed Central

    Esmaeili-Rad, Mohammad R.; Salahuddin, Sayeef

    2013-01-01

    One important use of layered semiconductors such as molybdenum disulfide (MoS2) could be in making novel heterojunction devices leading to functionalities unachievable using conventional semiconductors. Here we demonstrate a metal-semiconductor-metal heterojunction photodetector, made of MoS2 and amorphous silicon (a-Si), with rise and fall times of about 0.3 ms. The transient response does not show persistent (residual) photoconductivity, unlike conventional a-Si devices where it may last 3–5 ms, thus making this heterojunction roughly 10X faster. A photoresponsivity of 210 mA/W is measured at green light, the wavelength used in commercial imaging systems, which is 2−4X larger than that of a-Si and best reported MoS2 devices. The device could find applications in large area electronics, such as biomedical imaging, where a fast response is critical. PMID:23907598

  11. Peptide Bond Formation in Water Mediated by Carbon Disulfide.

    PubMed

    Leman, Luke J; Huang, Zheng-Zheng; Ghadiri, M Reza

    2015-09-01

    Demonstrating plausible nonenzymatic polymerization mechanisms for prebiotic monomers represents a fundamental goal in prebiotic chemistry. While a great deal is now known about the potentially prebiotic synthesis of amino acids, our understanding of abiogenic polymerization processes to form polypeptides is less well developed. Here, we show that carbon disulfide (CS2), a component of volcanic emission and sulfide mineral weathering, and a widely used synthetic reagent and solvent, promotes peptide bond formation in modest yields (up to ∼20%) from α-amino acids under mild aqueous conditions. Exposure of a variety of α-amino acids to CS2 initially yields aminoacyl dithiocarbamates, which in turn generate reactive 2-thiono-5-oxazolidone intermediates, the thio analogues of N-carboxyanhydrides. Along with peptides, thiourea and thiohydantoin species are produced. Amino acid stereochemistry was preserved in the formation of peptides. Our findings reveal that CS2 could contribute to peptide bond formation, and possibly other condensation reactions, in abiogenic settings. PMID:26308392

  12. Insulator-metal transition of highly compressed carbon disulfide

    SciTech Connect

    Dias, Ranga P.; Yoo, Choong-Shik; Kim, Minseob; Tse, John S.

    2012-04-24

    We present integrated spectral, structural, resistance, and theoretical evidences for simple molecular CS{sub 2} transformations to an insulating black polymer with threefold carbon atoms at 9 GPa, then to a semiconducting polymer above 30 GPa, and finally to a metallic solid above 50 GPa. The metallic phase is a highly disordered three-dimensional network structure with fourfold carbon atoms at the carbon-sulfur distance of {approx}1.70 {angstrom}. Based on first-principles calculations, we present two plausible structures for the metallic phase: {alpha}-chalcopyrite and tridymite, both of which exhibit metallic ground states and disordered diffraction features similar to that measured. We also present the phase and chemical transformation diagram for carbon disulfide, showing a large stability field of the metallic phase to 100 GPa and 800 K.

  13. On the Formation of Cometary Carbon Disulfide (CS2)

    NASA Technical Reports Server (NTRS)

    Hudson, Reggie; Moore, marla; Ferrante, Robert F.

    2009-01-01

    The formation of cometary CS molecules from carbon disulfide, CS2 , was proposed about 20 years before the latter's detection in comet 122P/de Vico by Jackson et al. (2002). However, the origin of CS2 has received little attention from either experimentalists or theorists. As part of our on-going laboratory program to investigate cometary molecules we have examined chemical reactions that lead to CS2 in the solid state. Icy mixtures of known cometary molecules were proton irradiated near 10 K to doses of several eV per molecule. Mid-IR spectroscopy was used as an in situ probe to record both CS2 formation in the ices and the destruction of precursors. We find that the most likely route to cometary CS2 is through OCS by way of the S + CO reaction.

  14. Wet chemical thinning of molybdenum disulfide down to its monolayer

    SciTech Connect

    Amara, Kiran Kumar; Chu, Leiqiang; Kumar, Rajeev; Toh, Minglin; Eda, Goki

    2014-09-01

    We report on the preparation of mono- and bi-layer molybdenum disulfide (MoS{sub 2}) from a bulk crystal by facile wet chemical etching. We show that concentrated nitric acid (HNO{sub 3}) effectively etches thin MoS{sub 2} crystals from their edges via formation of MoO{sub 3}. Interestingly, etching of thin crystals on a substrate leaves behind unreacted mono- and bilayer sheets. The flakes obtained by chemical etching exhibit electronic quality comparable to that of mechanically exfoliated counterparts. Our findings indicate that the self-limiting chemical etching is a promising top-down route to preparing atomically thin crystals from bulk layer compounds.

  15. Temperature effect on optical spectra of monolayer molybdenum disulfide

    NASA Astrophysics Data System (ADS)

    Soklaski, Ryan; Liang, Yufeng; Yang, Li

    2014-05-01

    Recently, measured optical absorption and photoluminescence spectra reveal substantial frequency shifts of both exciton and trion peaks as monolayer molybdenum disulfide, MoS2, is cooled from 363 K to 4 K. First-principles simulations using the GW-Bethe-Salpeter equation approach satisfactorily reproduce these frequency shifts by incorporating the thermal expansion effect. Studying these temperature effects in monolayer MoS2 is crucial for rectifying the results of available experiments with the previous predictions of zero-temperature-limit simulations. Moreover, our estimated thermal expansion coefficient of monolayer MoS2 is less than that of bulk counterpart by tracking the frequency shifts of the exciton peaks in optical spectra. This may serve as a convenient way to estimate thermal expansion coefficients of general two-dimensional chalcogenides.

  16. Disulfide bonds regulate binding of exogenous ligand to human cytoglobin.

    PubMed

    Tsujino, Hirofumi; Yamashita, Taku; Nose, Azusa; Kukino, Kaori; Sawai, Hitomi; Shiro, Yoshitsugu; Uno, Tadayuki

    2014-06-01

    Cytoglobin (Cgb) was discovered a decade ago and is a fourth member of the group of hexacoordinated globin-folded proteins. Although some crystal structures have been reported and several functions have been proposed for Cgb, its physiological role remains uncertain. In this study, we measured cyanide binding to the ferric state of the wild-type (WT) Cgb, and found that the binding consisted of multiple steps. These results indicated that Cgb may be comprised of several forms, and the presence of monomers, dimers, and tetramers was subsequently confirmed by SDS-PAGE. Remarkably, each species contained two distinguishable forms, and, in the monomer, analyses of alternative cysteine states suggested the presence of an intramolecular disulfide bond (monomer SS form) and a structure with unpaired thiol groups (monomer SH form). These confirmed that forms were separated by gel-exclusion chromatography, and that the cyanide binding of the separated fractions was again measured; they showed different affinities for cyanide, with the monomer fraction showing the highest affinity. In addition, the ferrous state in each fraction showed distinct carbon monoxide (CO)-binding properties, and the affinities for cyanide and CO suggested a linear correlation. Furthermore, we also prepared several variants involving the two cysteine residues. The C38S and C83S variants showed a binding affinity for cyanide similar to the value for the monomer SH form, and hence the fraction with the highest affinity for exogenous ligands was designated as a monomer SS form. We concluded that polymerization could be a mechanism that triggers the exertion of various physiological functions of this protein and that an appropriate disulfide bond between the two cysteine residues was critical for regulating the binding affinity of Cgb, which can act as a ROS scavenger, for exogenous ligands. PMID:24632414

  17. Exploiting scientific advances. Philip Russell.

    PubMed

    1992-04-01

    The Children's Vaccine Initiative (CVI) will hopefully accelerate the vaccine development process, make it more efficient, and produce new and better vaccines which will prevent most, if not all, of today's preventable diseases which lead to childhood mortality. The technology exists, but has simply not been exploited. Many exciting approaches to vaccine development never advance beyond the product development stage because, until now, there has been no mechanism for overseeing the entire process from the initial conception of a vaccine in the laboratory to its development by industry and its incorporation into vaccine programs. The CVI, however, has been established to provide such oversight and to coordinate the process. Recently developed technologies which could advance the attainment of CVI goals are the microencapsulation process and the use of live viral or attenuated bacterial vectors, genetically engineered to express desired vaccine antigen structures and induce immunity to specific infectious agents. The scientific obstacles are simply challenges which can be overcome. However, for the CVI to achieve its goals, it requires both adequate public sector resources and the collaboration of private industry. PMID:12321835

  18. Exploiting dual otoacoustic emission sources

    NASA Astrophysics Data System (ADS)

    Abdala, Carolina; Kalluri, Radha

    2015-12-01

    Two distinct processes generate otoacoustic emissions (OAEs). Reflection-source emissions, here recorded as stimulus frequency OAEs, are optimally informative at low sound levels and are more sensitive to slight hearing loss; they have been linked to cochlear amplifier gain and tuning. Distortion-source emissions are strongest at moderate-high sound levels and persist despite mild hearing loss; they likely originate in the nonlinear process of hair cell transduction. In this preliminary study, we exploit the unique features of each by generating a combined reflection-distortion OAE profile in normal hearing and hearing-impaired ears. Distortion-product (DP) and stimulus-frequency (SF) OAEs were recorded over a broad range of stimulus levels and frequencies. Individual I/O and transfer functions were generated for both emission types in each ear, and OAE peak strength, compression threshold, and rate of compression were calculated. These combined SFOAE and DPOAE features in normal and hearing-impaired ears may provide a potentially informative and novel index of hearing loss. This is an initial step toward utilizing OAE source in characterizing cochlear function and dysfunction.

  19. The Gaia scientific exploitation networks

    NASA Astrophysics Data System (ADS)

    Figueras, F.; Jordi, C.

    2015-05-01

    On July 2014 the Gaia satellite, placed at L2 since January 2014, finished their commissioning phase and started collecting high accurate scientific data. New and more realistic estimations of the astrometric, photometric and spectroscopic accuracy expected after five years mission operation (2014-2019) have been recently published in the Gaia Science Performance Web page. Here we present the coordination efforts and the activities being conducted through the two GREAT (Gaia Research for European Astronomy Training) European Networks, the GREAT-ESF, a programme supported by the European Science Foundation (2010-2015), and the GREAT-ITN network, from the European Union's Seventh Framework Programme (2011-2015). The main research theme of these networks is to unravel the origin and history of our home galaxy. Emphasis is placed on the research projects being conducted by the Spanish Researchers through these networks, well coordinated by the Red Española de Explotación Científica de Gaia (REG network, with more than 140 participants). Members of the REG play an important role on the collection of complementary spectroscopic data from ground based telescopes, on the development of new tools for an optimal scientific exploitation of Gaia data and on the preparation task to create the Gaia archive.

  20. Assistive Devices

    MedlinePlus

    ... center provides information on VA benefits for assistive technology. Medicare − Benefits may include assistive devices, such as ... a Web site that provides information about assistive technology products. Go to the “Products” section to find ...

  1. Two questions about surrogacy and exploitation.

    PubMed

    Wertheimer, Alan

    1992-01-01

    In this article I will consider two related questions about surrogacy and exploitation: (1) Is surrogacy exploitative? (2) If surrogacy is exploitative, what is the moral force of this exploitation? Briefly stated, I shall argue that whether surrogacy is exploitative depends on whether exploitation must be harmful to the exploited party or whether (as I think) there can be mutually advantageous exploitation. It also depends on some facts about surrogacy about which we have little reliable evidence and on our philosophical view on what counts as a harm to the surrogate. Our answer to the second question will turn in part on the account of exploitation we invoke in answering the first question and in part on the way in which we resolve some other questions about the justification of state interference. I shall suggest, however, that if surrogacy is a form of voluntary and mutually advantageous exploitation, then there is a strong presumption that surrogacy contracts should be permitted and even enforceable, although that presumption may be overridden on other grounds. PMID:11652070

  2. Direct measurement of the tryptophan-mediated photocleavage kinetics of a protein disulfide bond.

    PubMed

    Abaskharon, Rachel M; Gai, Feng

    2016-04-14

    Disulfide cleavage is one of the major causes underlying ultraviolet (UV) light-induced protein damage. While previous studies have provided strong evidence to support the notion that this process is mediated by photo-induced electron transfer from the excited state of an aromatic residue (e.g., tryptophan) to the disulfide bond, many mechanistic details are still lacking. For example, we do not know how quickly this process occurs in a protein environment. Herein, we design an experiment, which uses the unfolding kinetics of a protein as an observable, to directly assess the kinetics and mechanism of photo-induced disulfide cleavage. Our results show that this disulfide bond cleavage event takes place in ∼2 μs via a mechanism involving electron transfer from the triplet state of a tryptophan (Trp) residue to the disulfide bond. Furthermore, we find that one of the photoproducts of this reaction, a Trp-SR adduct, is formed locally, thus preventing the protein from re-cross-linking. Taken together, these findings suggest that a Trp-disulfide pair could be used as a photo-trigger to initiate protein folding dynamics and control the biological activities of disulfide-containing peptides. PMID:26997094

  3. Evaluation of protein disulfide conversion in vitro using a continuous flow dialysis system.

    PubMed

    Jiang, Xinzhao Grace; Wang, Tian; Kaltenbrunner, Oliver; Chen, Kenneth; Flynn, Gregory C; Huang, Gang

    2013-01-15

    Recombinant therapeutic proteins are heterogeneous due to chemical and physical modifications. Understanding the impact of these modifications on drug safety and efficacy is critical for optimal process development and for setting reasonable specification limits. In this study, we describe the development of an in vitro continuous flow dialysis system to evaluate potential in vivo behavior of thiol adducted species and incorrectly disulfide bonded species of therapeutic proteins. The system is capable of maintaining the low-level cysteine concentrations found in human blood. Liabilities of cysteamine adducted species, incorrectly disulfide bonded species, and the correctly disulfide bonded form of an Fc-fusion protein were studied using this system. Results showed that 90% of the cysteamine adduct converted into the correctly disulfide bonded form and incorrectly disulfide bonded species in approximately 4 h under physiological conditions. Approximately 50% of incorrectly disulfide bonded species converted into the correctly bonded form in 2 days. These results provide valuable information on potential in vivo stability of the cysteamine adduct, incorrectly disulfide bonded species, and the correctly bonded form of the Fc-fusion protein. These are important considerations when evaluating the criticality of product quality attributes. PMID:23022378

  4. Human β-Defensin 4 with Non-Native Disulfide Bridges Exhibit Antimicrobial Activity

    PubMed Central

    Sharma, Himanshu; Nagaraj, Ramakrishnan

    2015-01-01

    Human defensins play multiple roles in innate immunity including direct antimicrobial killing and immunomodulatory activity. They have three disulfide bridges which contribute to the stability of three anti-parallel β-strands. The exact role of disulfide bridges and canonical β-structure in the antimicrobial action is not yet fully understood. In this study, we have explored the antimicrobial activity of human β-defensin 4 (HBD4) analogs that differ in the number and connectivity of disulfide bridges. The cysteine framework was similar to the disulfide bridges present in μ-conotoxins, an unrelated class of peptide toxins. All the analogs possessed enhanced antimicrobial potency as compared to native HBD4. Among the analogs, the single disulfide bridged peptide showed maximum potency. However, there were no marked differences in the secondary structure of the analogs. Subtle variations were observed in the localization and membrane interaction of the analogs with bacteria and Candida albicans, suggesting a role for disulfide bridges in modulating their antimicrobial action. All analogs accumulated in the cytosol where they can bind to anionic molecules such as nucleic acids which would affect several cellular processes leading to cell death. Our study strongly suggests that native disulfide bridges or the canonical β-strands in defensins have not evolved for maximal activity but they play important roles in determining their antimicrobial potency. PMID:25785690

  5. Structural stability of human alpha-thrombin studied by disulfide reduction and scrambling.

    PubMed

    Rajesh Singh, R; Chang, Jui Yoa

    2003-09-23

    Human alpha-thrombin is a very important plasma serine protease, which is involved in physiologically vital processes like hemostasis, thrombosis, and activation of platelets. Knowledge regarding the structural stability of alpha-thrombin is essential for understanding its biological regulation. Here, we investigated the structural and conformational stability of alpha-thrombin using the techniques of disulfide reduction and disulfide scrambling. alpha-Thrombin is composed of a light A-chain (36 residues) and a heavy B-chain (259 residues) linked covalently by an inter-chain disulfide bond (Cys(1)-Cys(122)). The B-chain is stabilized by three intra-chain disulfide bonds (Cys(42)-Cys(58), Cys(168)-Cys(182), and Cys(191)-Cys(220)) (Chymotrypsinogen nomenclature). Upon reduction with dithiothreitol (DTT), alpha-thrombin unfolded in a 'sequential' manner with sequential reduction of Cys(168)-Cys(182) within the B-chain followed by the inter-chain disulfide, generating two distinct partially reduced intermediates, I-1 and I-2, respectively. Conformational stability of alpha-thrombin was investigated by the technique of disulfide scrambling. alpha-Thrombin denatures by scrambling its native disulfide bonds in the presence of denaturant [urea, guanidine hydrochloride (GdmCl) or guanidine thiocyanate (GdmSCN)] and a thiol initiator. During the process, cleavage of the inter-chain disulfide bond and release of the A-chain from B-chain was the foremost event. The three disulfides in the B-chain subsequently scrambled to form three major isomers (designated as X-Ba, X-Bb, and X-Bc). Complete denaturation of alpha-thrombin was observed at low concentrations of denaturants (0.5 M GdmSCN, 1.5 M GdmCl, or 3 M urea) indicating low conformational stability of the protease. PMID:14499592

  6. Disulfide oil hazard assessment using categorical analysis and a mode of action determination.

    PubMed

    Morgott, David; Lewis, Christopher; Bootman, James; Banton, Marcy

    2014-01-01

    Diethyl and diphenyl disulfides, naphtha sweetening (Chemical Abstracts Service [CAS] # 68955-96-4), are primarily composed of low-molecular-weight dialkyl disulfides extracted from C4 to C5 light hydrocarbon streams during the refining of crude oil. The substance, commonly known as disulfide oil (DSO), can be composed of up to 17 different disulfides and trisulfides with monoalkyl chain lengths no greater than C4. The disulfides in DSO constitute a homologous series of chemical constituents that are perfectly suited for a hazard evaluation using a read-across/worst-case approach. The DSO constituents exhibit a common mode of action that is operable at all trophic levels. The observed oxidative stress response is mediated by reactive oxygen species and free radical intermediates generated after disulfide bond cleavage and subsequent redox cycling of the resulting mercaptan. Evidence indicates that the lowest series member, dimethyl disulfide (DMDS), can operate as a worst-case surrogate for other members of the series, since it displays the highest toxicity. Increasing the alkyl chain length or degree of substitution has been shown to serially reduce disulfide toxicity through resonance stabilization of the radical intermediate or steric inhibition of the initial enzymatic step. The following case study examines the mode of action for dialkyl disulfide toxicity and documents the use of read-across information from DMDS to assess the hazards of DSO. The results indicate that DSO possesses high aquatic toxicity, moderate environmental persistence, low to moderate acute toxicity, high repeated dose toxicity, and a low potential for genotoxicity, carcinogenicity, and reproductive/developmental effects. PMID:24189075

  7. Characterization of the kringle fold and identification of a ubiquitous new class of disulfide rotamers.

    PubMed

    Ozhogina, Olga A; Bominaar, Emile L

    2009-11-01

    The disulfide-bridged chains in the kringle (K) and fibronectin type II (FN2) domains are characterized using a taxonomy that considers the regularities in both beta-secondary structure and cystine cluster. The structural core of the kringle fold comprises an assembly of two beta-hairpins (a "beta-meander") accommodating two overlapping disulfides; one cystine is incorporated in adjacent beta-strands, whereas the other is located just beyond the ends of non-adjacent beta-strands. The dispositions of the (N, C) termini of the two overlapping disulfides in the kringle fold are given as (m, j+1) and (i-1, k+1), in which m, i, j, and k (mdisulfide kringle-cross". The metrics of this motif are quantified, revealing structural differences between the two families of the kringle fold. The conformations of disulfides in the kringle fold are poorly accommodated by existing classification schemes. To elucidate the nature of these rotamers we have performed density functional theory (DFT) calculations for diethyl disulfide. A new classification for the disulfide conformations in proteins is proposed, consisting of six rotamer types: spiral, trans-spiral, corner, trans, hook, and staple. Its relation with previous classification schemes is specified. A survey of high-resolution X-ray structures reveals that the disulfide conformations are clustered around the averaged conformations for the six classes. Average conformation dihedral and distance values are in excellent agreement with the DFT values. The two overlapping disulfides in kringle domains adopt the trans-spiral conformation that appears to be ubiquitous (~17%) in proteins. One of the disulfides stretches across the beta

  8. MS2DB+: a software for determination of disulfide bonds using multi-ion analysis.

    PubMed

    Murad, William; Singh, Rahul

    2013-06-01

    MS2DB+ is an open-source platform-independent web application for determining, in polynomial time, the disulfide linkages in proteins using tandem mass spectrometry (MS/MS) data. It utilizes an efficient approximation algorithm which allows the consideration of multiple ion-types (a, a(o), a*, b, b(o), b*, c, x, y, y(o), y*, and z) in the analysis. Once putative disulfide bonds are identified, a graph optimization approach is used to obtain the most likely global disulfide connectivity pattern. PMID:23096131

  9. Kinetics and intracellular location of intramolecular disulfide bond formation mediated by the cytoplasmic redox system encoded by vaccinia virus

    SciTech Connect

    Bisht, Himani; Brown, Erica; Moss, Bernard

    2010-03-15

    Poxviruses encode a redox system for intramolecular disulfide bond formation in cytoplasmic domains of viral proteins. Our objectives were to determine the kinetics and intracellular location of disulfide bond formation. The vaccinia virus L1 myristoylated membrane protein, used as an example, has three intramolecular disulfide bonds. Reduced and disulfide-bonded forms of L1 were distinguished by electrophoretic mobility and reactivity with monoclonal and polyclonal antibodies. Because disulfide bonds formed during 5 min pulse labeling with radioactive amino acids, a protocol was devised in which dithiothreitol was present at this step. Disulfide bond formation was detected by 2 min after removal of reducing agent and was nearly complete in 10 min. When the penultimate glycine residue was mutated to prevent myristoylation, L1 was mistargeted to the endoplasmic reticulum and disulfide bond formation failed to occur. These data suggested that viral membrane association was required for oxidation of L1, providing specificity for the process.

  10. Combined use of ion mobility and collision-induced dissociation to investigate the opening of disulfide bridges by electron-transfer dissociation in peptides bearing two disulfide bonds.

    PubMed

    Massonnet, Philippe; Upert, Gregory; Smargiasso, Nicolas; Gilles, Nicolas; Quinton, Loïc; De Pauw, Edwin

    2015-01-01

    Disulfide bonds are post-translational modifications (PTMs) often found in peptides and proteins. They increase their stability toward enzymatic degradations and provide the structure and (consequently) the activity of such folded proteins. The characterization of disulfide patterns, i.e., the cysteine connectivity, is crucial to achieve a global picture of the active conformation of the protein of interest. Electron-transfer dissociation (ETD) constitutes a valuable tool to cleave the disulfide bonds in the gas phase, avoiding chemical reduction/alkylation in solution. To characterize the cysteine pairing, the present work proposes (i) to reduce by ETD one of the two disulfide bridges of model peptides, resulting in the opening of the cyclic structures, (ii) to separate the generated species by ion mobility, and (iii) to characterize the species using collision-induced dissociation (CID). Results of this strategy applied to several peptides show different behaviors depending on the connectivity. The loss of SH· radical species, observed for all the peptides, confirms the cleavage of the disulfides during the ETD process. PMID:25915795