sulfhydryl reagents


Summary: Chemical agents that react with SH groups. This is a chemically diverse group that is used for a variety of purposes. Among these are enzyme inhibition, enzyme reactivation or protection, and labelling.

Top Publications

  1. Cui Y, Wang W, Fan Z. Cytoplasmic vestibule of the weak inward rectifier Kir6.2 potassium channel. J Biol Chem. 2002;277:10523-30 pubmed
    ..Data are consistent with the hypothesis that residues Ile-210-Ser-212 line a funnel-shaped vestibule of 20-25 A in diameter, which remains unchanged during channel gating. ..
  2. Zhou Y, Kanner B. Transporter-associated currents in the gamma-aminobutyric acid transporter GAT-1 are conditionally impaired by mutations of a conserved glycine residue. J Biol Chem. 2005;280:20316-24 pubmed
    ..Our results can be explained by a model invoking two outward-facing states of the empty transporter and a defective transition between these states in the glycine 80 mutants. ..
  3. Nakunst D, Larisch C, Hüser A, Tauch A, Pühler A, Kalinowski J. The extracytoplasmic function-type sigma factor SigM of Corynebacterium glutamicum ATCC 13032 is involved in transcription of disulfide stress-related genes. J Bacteriol. 2007;189:4696-707 pubmed
    ..A typical SigH-responsive promoter was mapped upstream of the sigM gene. The ECF sigma factor SigM is apparently part of a regulatory cascade, and its transcription is controlled by SigH under conditions of thiol-oxidative stress...
  4. Zhen X, Xie C, Fitzmaurice A, Schoonover C, Orenstein E, Yang J. Functional architecture of the inner pore of a voltage-gated Ca2+ channel. J Gen Physiol. 2005;126:193-204 pubmed
    ..This work provides an impetus for future studies on ion permeation, gating, and drug binding of VGCCs. ..
  5. Bruening Wright A, Schumacher M, Adelman J, Maylie J. Localization of the activation gate for small conductance Ca2+-activated K+ channels. J Neurosci. 2002;22:6499-506 pubmed
    ..The results suggest that the SK channel gate is not formed by the cytoplasmic end of S6 but resides deep in the channel pore in or near the selectivity filter. ..
  6. Bell D, Yao H, Saenger R, Riley J, Siegelbaum S. Changes in local S4 environment provide a voltage-sensing mechanism for mammalian hyperpolarization-activated HCN channels. J Gen Physiol. 2004;123:5-19 pubmed
  7. Rowland M, Tsigelny I, Wolfe M, Pezzementi L. Inactivation of an invertebrate acetylcholinesterase by sulfhydryl reagents: a reconsideration of the implications for insecticide design. Chem Biol Interact. 2008;175:73-5 pubmed publisher
    ..the inactivation of an invertebrate acetylcholinesterase, cholinesterase 2 from amphioxus, by the sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM)...
  8. Nakajo K, Kubo Y. KCNE1 and KCNE3 stabilize and/or slow voltage sensing S4 segment of KCNQ1 channel. J Gen Physiol. 2007;130:269-81 pubmed
    ..These results offer new insight into the mechanism of KCNQ1 channel modulation by KCNE1 and KCNE3. ..
  9. Pezzementi L, Rowland M, Wolfe M, Tsigelny I. Inactivation of an invertebrate acetylcholinesterase by sulfhydryl reagents: the roles of two cysteines in the catalytic gorge of the enzyme. Invert Neurosci. 2006;6:47-55 pubmed investigate the inactivation of an invertebrate acetylcholinesterase (AChE), ChE2 from amphioxus, by the sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM), creating various mutants, including ..

More Information


  1. Xie C, Zhen X, Yang J. Localization of the activation gate of a voltage-gated Ca2+ channel. J Gen Physiol. 2005;126:205-12 pubmed
    ..Our results suggest that the S6 helices of the alpha1 subunit of VGCCs undergo conformation changes during gating and the activation gate is located at the intracellular end of the pore. ..
  2. Nabemoto M, Ohsawa K, Nakamura H, Hirabayashi T, Saito T, Okuma Y, et al. Reversible activation of secretory phospholipase A2 by sulfhydryl reagents. Arch Biochem Biophys. 2005;436:145-53 pubmed
    ..Both sulfhydryl reagents stimulated AA release from the HEK293T cells expressing human sPLA(2)X, and stimulated the sPLA(2) ..
  3. Capobianco L, Impagnatiello T, Ferramosca A, Zara V. The mitochondrial tricarboxylate carrier of silver eel: chemical modification by sulfhydryl reagents. J Biochem Mol Biol. 2004;37:515-21 pubmed
    ..Incubation of the proteoliposomes with various sulfhydryl reagents led to inhibition of the reconstituted citrate transport activity...
  4. Zomot E, Kanner B. The interaction of the gamma-aminobutyric acid transporter GAT-1 with the neurotransmitter is selectively impaired by sulfhydryl modification of a conformationally sensitive cysteine residue engineered into extracellular loop IV. J Biol Chem. 2003;278:42950-8 pubmed
    ..accessible cysteine residue of the wild-type has been replaced by alanine, is also highly sensitive to the sulfhydryl reagents. Maximal sensitivity is observed in the presence of the cosubstrates sodium and chloride...
  5. Canfield L. Vitamin K-dependent oxygenase/carboxylase; differential inactivation by sulfhydryl reagents. Biochem Biophys Res Commun. 1987;148:184-91 pubmed
    Inhibition of vitamin K-dependent carboxylase and oxygenase by sulfhydryl reagents was compared...
  6. Larsson H, Baker O, Dhillon D, Isacoff E. Transmembrane movement of the shaker K+ channel S4. Neuron. 1996;16:387-97 pubmed
    ..These results demonstrate that the putative voltage-sensing charges of S4 actually reside in the membrane and that they move outward when channels open. We consider constraints placed on channel structure by these results. ..
  7. Yamaoka K, Yakehiro M, Yuki T, Fujii H, Seyama I. Effect of sulfhydryl reagents on the regulatory system of the L-type Ca channel in frog ventricular myocytes. Pflugers Arch. 2000;440:207-15 pubmed
    ..It is concluded that there are sites on the Ca channels that are subject to direct modification by SH reagents...
  8. Wu X, Edwards H, Sather W. Side chain orientation in the selectivity filter of a voltage-gated Ca2+ channel. J Biol Chem. 2000;275:31778-85 pubmed
    ..Thus the structure of the Ca(2+) channel selectivity filter differs substantially from that of ancestral K(+) channels...
  9. Roux M, Martínez Maza R, Le Goff A, Lopez Corcuera B, Aragon C, Supplisson S. The glial and the neuronal glycine transporters differ in their reactivity to sulfhydryl reagents. J Biol Chem. 2001;276:17699-705 pubmed
    ..Together, these results indicate that EL1 conformation differs between GlyT1b and GlyT2a and is modified by substrate binding and translocation. ..
  10. Yanamandra S, Sarrafee S, Anaya Bergman C, Jones K, Lewis J. Role of the Porphyromonas gingivalis extracytoplasmic function sigma factor, SigH. Mol Oral Microbiol. 2012;27:202-19 pubmed publisher
    ..Collectively, our studies demonstrate that SigH is a positive regulator of gene expression required for survival of the bacterium in the presence of oxygen and oxidative stress, hemin uptake and virulence. ..
  11. Cederholm J, Absalom N, Sugiharto S, Griffith R, Schofield P, Lewis T. Conformational changes in extracellular loop 2 associated with signal transduction in the glycine receptor. J Neurochem. 2010;115:1245-55 pubmed publisher
    ..These results demonstrate that loop 2 does undergo a conformational change as part of the mechanism that couples ligand binding to channel opening. ..
  12. Vöpel T, Kunzelmann S, Herrmann C. Nucleotide dependent cysteine reactivity of hGBP1 uncovers a domain movement during GTP hydrolysis. FEBS Lett. 2009;583:1923-7 pubmed publisher
    ..Our results indicate that nucleotide hydrolysis induces a domain movement in hGBP1, which we suggest enables further assembly of the protein. ..
  13. Hepp S, Muller M. Sulfhydryl oxidation: a potential strategy to achieve neuroprotection during severe hypoxia?. Neuroscience. 2008;152:903-12 pubmed publisher
    ..Attenuation of the severe changes during HSD or an improved outcome was not observed. Nevertheless, combination of SH-oxidants with established neuroprotectants might be a potential therapeutic approach. ..
  14. Sridevi K, Udgaonkar J. Unfolding rates of barstar determined in native and low denaturant conditions indicate the presence of intermediates. Biochemistry. 2002;41:1568-78 pubmed
  15. Vervoort E, Bultema J, Schuurman Wolters G, Geertsma E, Broos J, Poolman B. The first cytoplasmic loop of the mannitol permease from Escherichia coli is accessible for sulfhydryl reagents from the periplasmic side of the membrane. J Mol Biol. 2005;346:733-43 pubmed
    ..These and other results suggest that the predicted loop between TM2 and TM3 may fold back into the membrane and form part of the translocation path. ..
  16. Gupta K, Panda D. Perturbation of microtubule polymerization by quercetin through tubulin binding: a novel mechanism of its antiproliferative activity. Biochemistry. 2002;41:13029-38 pubmed
    ..The data suggest that quercetin inhibits cancer cells proliferation at least in part by perturbing microtubule functions through tubulin binding. ..
  17. Pierre S, Guillot A, Benjdia A, Sandstrom C, Langella P, Berteau O. Thiostrepton tryptophan methyltransferase expands the chemistry of radical SAM enzymes. Nat Chem Biol. 2012;8:957-9 pubmed publisher
    ..We show that it uses unprecedented chemistry to methylate inactivated sp(2)-hybridized carbon atoms, despite being predicted to be a radical SAM enzyme. ..
  18. Hart E, Powers Lee S. Role of Cys-1327 and Cys-1337 in redox sensitivity and allosteric monitoring in human carbamoyl phosphate synthetase. J Biol Chem. 2009;284:5977-85 pubmed publisher
    ..These findings strongly suggested an alternative eCPS conformation relative to the single crystal conformation thus far identified. ..
  19. Camerini S, Polci M, Bachi A. Proteomics approaches to study the redox state of cysteine-containing proteins. Ann Ist Super Sanita. 2005;41:451-7 pubmed
    ..This methodology can be used in proteomic studies to recognize redox-sensitive Cysteine containing proteins and nitric oxide targets. ..
  20. Chraibi A, Horisberger J. Na self inhibition of human epithelial Na channel: temperature dependence and effect of extracellular proteases. J Gen Physiol. 2002;120:133-45 pubmed
    ..The extracellular Na(+)-dependent inactivation has a large energy of activation and can be abolished by treatment with extracellular proteases. ..
  21. Polyakova O, Roitel O, Asryants R, Poliakov A, Branlant G, Muronetz V. Misfolded forms of glyceraldehyde-3-phosphate dehydrogenase interact with GroEL and inhibit chaperonin-assisted folding of the wild-type enzyme. Protein Sci. 2005;14:921-8 pubmed
    ..Thus, we suggest that interaction of GroEL with certain types of misfolded proteins can result in the formation of stable complexes and the impairment of chaperonin activity. ..
  22. Labriola J, daCosta C, Wang S, Figeys D, Smith J, Sturgeon R, et al. Phospholipase C activity affinity purifies with the Torpedo nicotinic acetylcholine receptor. J Biol Chem. 2010;285:10337-43 pubmed publisher
    ..The acetylcholine receptor may directly or indirectly influence lipid metabolism in a manner that enhances its own function. ..
  23. Landino L, Hasan R, McGaw A, Cooley S, Smith A, Masselam K, et al. Peroxynitrite oxidation of tubulin sulfhydryls inhibits microtubule polymerization. Arch Biochem Biophys. 2002;398:213-20 pubmed
    ..5 to 6.3 mol cysteine/mol tubulin. These results indicate that cysteine oxidation of tubulin by peroxynitrite, rather than tyrosine nitration, is the primary mechanism of inhibition of microtubule polymerization. ..
  24. Kot M, Zaborska W. Irreversible inhibition of jack bean urease by pyrocatechol. J Enzyme Inhib Med Chem. 2003;18:413-7 pubmed
    ..These results proved that the urease active site was involved in the pyrocatechol inactivation. ..
  25. Goyal S. Effect of sample flow rate and sampling duration on the absorption of NO2 in the sodium arsenite monitoring method. J Environ Monit. 2002;4:579-82 pubmed
    ..Correction factors for the absorption efficiency were evolved that can be used to update the existing database, as well as future NO2 data generated using this manual monitoring method. ..
  26. Priora R, Coppo L, Margaritis A, Di Giuseppe D, Frosali S, Summa D, et al. The control of S-thiolation by cysteine via gamma-glutamyltranspeptidase and thiol exchanges in erythrocytes and plasma of diamide-treated rats. Toxicol Appl Pharmacol. 2010;242:333-43 pubmed publisher
    ..Moreover, reduced cysteine was found to be involved in deglutathionylation processes. Mechanisms of protein glutathionylation by diamide and deglutathionylation by cysteine were proposed. ..
  27. Lee K, Shimoji M, Hossain Q, Sunakawa H, Aniya Y. Novel function of glutathione transferase in rat liver mitochondrial membrane: role for cytochrome c release from mitochondria. Toxicol Appl Pharmacol. 2008;232:109-18 pubmed publisher
    ..Furthermore, mtMGST1 activity was also inhibited by CsA and BKA. These results indicate that mtMGST1 is activated through mixed disulfide bond formation that contributes to cytochrome c release from mitochondria through the MPT pore. ..
  28. El Hiani Y, Linsdell P. Changes in accessibility of cytoplasmic substances to the pore associated with activation of the cystic fibrosis transmembrane conductance regulator chloride channel. J Biol Chem. 2010;285:32126-40 pubmed publisher
    ..One logical interpretation of these findings is that cytoplasmic access to residues at the narrowest region of the pore changes concomitant with activation. ..
  29. Li S, Kimura M, Takashima T, Hayashi K, Inoue K, Ishiguro R, et al. Role of cysteine residues in 4-oxalomesaconate hydratase from Pseudomonas ochraceae NGJ1. Biosci Biotechnol Biochem. 2007;71:449-57 pubmed
  30. Gong L, Gao T, Huang H, Zhuang Z, Tong Z. Transient forebrain ischemia induces persistent hyperactivity of large conductance Ca2+-activated potassium channels via oxidation modulation in rat hippocampal CA1 pyramidal neurons. Eur J Neurosci. 2002;15:779-83 pubmed
    ..The ischemia-induced augmentation in BKCa channel activity may be also associated with the postischemic neuronal injury. ..
  31. Xiao H, Zhou D, Liu S, Qi R, Zheng Y, Huang Y, et al. Delivery of active DACH-Pt anticancer species by biodegradable amphiphilic polymers using thiol-ene radical addition. Macromol Biosci. 2012;12:367-73 pubmed publisher
    ..In vitro studies show that the DACH-Pt micelles display enhanced or comparable cytotoxicity against SKOV-3 and MCF-7 cancer cells, while they show reduced toxicity to HeLa cells compared with oxaliplatin. ..
  32. Sakuma S, Fujita J, Nakanishi M, Wada S, Fujimoto Y. Disulfide S-monoxides convert xanthine dehydrogenase into oxidase in rat liver cytosol more potently than their respective disulfides. Biol Pharm Bull. 2008;31:1013-6 pubmed
    ..These results suggest that disulfide S-monoxides have the potential to increase the generation of reactive oxygen species through the conversion of XD to XO in liver. ..
  33. Chang C, Simplaceanu V, Ho C. Effects of amino acid substitutions at beta 131 on the structure and properties of hemoglobin: evidence for communication between alpha 1 beta 1- and alpha 1 beta 2-subunit interfaces. Biochemistry. 2002;41:5644-55 pubmed
  34. Ishima Y, Akaike T, Kragh Hansen U, Hiroyama S, Sawa T, Suenaga A, et al. S-nitrosylated human serum albumin-mediated cytoprotective activity is enhanced by fatty acid binding. J Biol Chem. 2008;283:34966-75 pubmed publisher
    ..A small nitric oxide transfer takes place in a slow system, which is unaffected by fatty acid binding to SNO-HSA and not influenced by filipin III. Thus, fatty acids could be a novel type of mediator for S-transnitrosation. ..
  35. Kulkarni A, Haworth I, Lee V. Transmembrane segment 5 of the dipeptide transporter hPepT1 forms a part of the substrate translocation pathway. Biochem Biophys Res Commun. 2003;306:177-85 pubmed
  36. Reott M, Parker A, Rocha E, Smith C. Thioredoxins in redox maintenance and survival during oxidative stress of Bacteroides fragilis. J Bacteriol. 2009;191:3384-91 pubmed publisher
    ..These data provide insight into the role of individual Trxs in the B. fragilis oxidative stress response. ..
  37. Yu Y, Jia P, Huang Y, Cai X, Chen G. [Diamide and cyclosporin A enhanced arsenic trioxide-induced apoptosis in NB4 cells]. Zhonghua Xue Ye Xue Za Zhi. 2002;23:254-7 pubmed
    ..7% and 42.2%, respectively. As(2)O(3)-induced DeltaPsim disruption possibly involves with thiol oxidation or crosslink of important components especially ANT-related molecules. ..
  38. Mima J, Suzuki H, Takahashi M, Hayashi R. Overexpression and functional characterization of a serine carboxypeptidase inhibitor (I(C)) from Saccharomyces cerevisiae. J Biochem. 2002;132:967-73 pubmed
    ..0 x 10(-9) M. The chemical modification of I(C) with sulfhydryl reagents indicated that it lacks disulfide bonds and has two free SH groups, which are responsible, not for the ..
  39. May J, Li L, Qu Z, Cobb C. Mitochondrial recycling of ascorbic acid as a mechanism for regenerating cellular ascorbate. Biofactors. 2007;30:35-48 pubmed
  40. Baty J, Hampton M, Winterbourn C. Detection of oxidant sensitive thiol proteins by fluorescence labeling and two-dimensional electrophoresis. Proteomics. 2002;2:1261-6 pubmed
    ..Separation of proteins by 2-D electrophoresis revealed approximately 200 thiol proteins that were oxidized by diamide treatment. This method will be valuable in elucidating redox signaling pathways. ..
  41. Gallardo N, Arribas C, Villar M, Ros M, Carrascosa J, Martinez C, et al. ObRa and ObRe are differentially expressed in adipose tissue in aged food-restricted rats: effects on circulating soluble leptin receptor levels. Endocrinology. 2005;146:4934-42 pubmed
    ..These data suggest that shedding of ObRa membrane-anchored receptors, rather than ObRe expression, might preferentially contribute to the generation of the increased levels of SLR in serum under conditions of FR. ..
  42. Liapakis G, Simpson M, Javitch J. The substituted-cysteine accessibility method (SCAM) to elucidate membrane protein structure. Curr Protoc Neurosci. 2001;Chapter 4:Unit 4.15 pubmed publisher
    ..The protocol in this unit describes the use of SCAM to map the binding-site crevice of a G-protein coupled receptor (GPCR) which binds ligand within the transmembrane portion of the receptor. ..
  43. Zhang Z, Xu Y, Dong P, Sharma D, Chiamvimonvat N. A negatively charged residue in the outer mouth of rat sodium channel determines the gating kinetics of the channel. Am J Physiol Cell Physiol. 2003;284:C1247-54 pubmed
    ..Our data represent the first report indicating the role of negative charge in the slow inactivation of the voltage-gated Na(+) channel. ..
  44. Olschewski A, Hong Z, Peterson D, Nelson D, Porter V, Weir E. Opposite effects of redox status on membrane potential, cytosolic calcium, and tone in pulmonary arteries and ductus arteriosus. Am J Physiol Lung Cell Mol Physiol. 2004;286:L15-22 pubmed
    ..These observations support the concept that redox changes could signal the opposite effects of oxygen in the PA and DA. ..
  45. Lozinsky E, Febbraio F, Shames A, Likhtenshtein G, Bismuto E, Nucci R. Heterogeneity in the structural dynamics of Sulfolobus solfataricus beta-glycosidase revealed by electron paramagnetic resonance and frequency domain fluorometry. Protein Sci. 2002;11:2535-44 pubmed
    ..e., 328 K and 338 K, respectively). The Arrhenius analysis of data provided the activation energy relative to the conformational changes characterizing the local and global movements running through the protein matrix. ..
  46. Shubenok D, Tsybovsky Y, Stremovskiy O, Deyev S, Martsev S. Fusion of barnase to antiferritin antibody F11 VH domain results in a partially folded functionally active protein. Biochemistry (Mosc). 2009;74:672-80 pubmed
  47. Pallis M, Bradshaw T, Westwell A, Grundy M, Stevens M, Russell N. Induction of apoptosis without redox catastrophe by thioredoxin-inhibitory compounds. Biochem Pharmacol. 2003;66:1695-705 pubmed
    ..Phenylarsine oxide induced both free radicals and hydrogen peroxide, but this did not appear to interfere with apoptosis. We conclude that pharmacological targeting of thioredoxin can induce a well-orchestrated apoptotic programme. ..
  48. Abramić M, Simaga S, Osmak M, Cicin Sain L, Vukelić B, Vlahovicek K, et al. Highly reactive cysteine residues are part of the substrate binding site of mammalian dipeptidyl peptidases III. Int J Biochem Cell Biol. 2004;36:434-46 pubmed
    ..These results provide kinetic evidence that cysteine residues are involved in substrate binding of mammalian DPPs III. ..
  49. Steenkamp D, Vogt R. Preparation and utilization of a reagent for the isolation and purification of low-molecular-mass thiols. Anal Biochem. 2004;325:21-7 pubmed
    ..Following their isolation as the heterodisulfides the thiol species of interest are recovered by reduction and facile separation from the apolar 2-mercapto-6-hydroxynaphthalene partner. ..
  50. Seki M, Awata J, Shimada K, Kashiyama T, Ito K, Yamamoto K. Susceptibility of Chara myosin to SH reagents. Plant Cell Physiol. 2003;44:201-5 pubmed
    ..corallina decreased to a level accountable for the decrease in the velocity of cytoplasmic streaming but it was also found that Chara myosin was far less susceptible to NEM than skeletal muscle myosin. ..
  51. Holmgren A, Sengupta R. The use of thiols by ribonucleotide reductase. Free Radic Biol Med. 2010;49:1617-28 pubmed publisher
    ..The involvement of cysteine residues in catalysis is common to all three classes of RNR. Taking account of the recent progress in this field of research, this review focuses on the use of thiols in the redox mechanism of RNR enzymes. ..
  52. Cappello A, Miniero D, Curcio R, Ludovico A, Daddabbo L, Stipani I, et al. Functional and structural role of amino acid residues in the odd-numbered transmembrane alpha-helices of the bovine mitochondrial oxoglutarate carrier. J Mol Biol. 2007;369:400-12 pubmed
    ..The transport activity of the mutant carriers was measured in the presence and absence of sulfhydryl reagents. The observed effects were rationalized by using a comparative structural model of the OGC...
  53. Kim Choi E, Danilo C, Kelly J, Carroll R, Shonnard D, Rybina I. Kinetic characterization and in vitro toxicity evaluation of a luciferase less susceptible to HPV chemical inhibition. Toxicol In Vitro. 2006;20:1537-47 pubmed
    ..The performance of CNBLuc03-06 in cell-based in vitro toxicity assays was shown to be superior to the current commercial formulation. ..