Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

Science 320 (5879): 1050-1054

Copyright © 2008 by the American Association for the Advancement of Science

Regulated Protein Denitrosylation by Cytosolic and Mitochondrial Thioredoxins

Moran Benhar,1 Michael T. Forrester,2 Douglas T. Hess,1 Jonathan S. Stamler1,2*

Abstract: Nitric oxide acts substantially in cellular signal transduction through stimulus-coupled S-nitrosylation of cysteine residues. The mechanisms that might subserve protein denitrosylation in cellular signaling remain uncharacterized. Our search for denitrosylase activities focused on caspase-3, an exemplar of stimulus-dependent denitrosylation, and identified thioredoxin and thioredoxin reductase in a biochemical screen. In resting human lymphocytes, thioredoxin-1 actively denitrosylated cytosolic caspase-3 and thereby maintained a low steady-state amount of S-nitrosylation. Upon stimulation of Fas, thioredoxin-2 mediated denitrosylation of mitochondria-associated caspase-3, a process required for caspase-3 activation, and promoted apoptosis. Inhibition of thioredoxin-thioredoxin reductases enabled identification of additional substrates subject to endogenous S-nitrosylation. Thus, specific enzymatic mechanisms may regulate basal and stimulus-induced denitrosylation in mammalian cells.

1 Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
2 Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.

* To whom correspondence should be addressed. E-mail: staml001{at}

Protein S-nitrosylation in preconditioning and postconditioning.
C. Penna, C. Angotti, and P. Pagliaro (2014)
Experimental Biology and Medicine
   Abstract »    Full Text »    PDF »
Thioredoxin-mediated Denitrosylation Regulates Cytokine-induced Nuclear Factor {kappa}B (NF-{kappa}B) Activation.
Z. T. Kelleher, Y. Sha, M. W. Foster, W. M. Foster, M. T. Forrester, and H. E. Marshall (2014)
J. Biol. Chem. 289, 3066-3072
   Abstract »    Full Text »    PDF »
Cysteine 96 of Ntcp is responsible for NO-mediated inhibition of taurocholate uptake.
U. Ramasamy, M. S. Anwer, and C. M. Schonhoff (2013)
Am J Physiol Gastrointest Liver Physiol 305, G513-G519
   Abstract »    Full Text »    PDF »
Regulation of Protein Function and Signaling by Reversible Cysteine S-Nitrosylation.
N. Gould, P.-T. Doulias, M. Tenopoulou, K. Raju, and H. Ischiropoulos (2013)
J. Biol. Chem. 288, 26473-26479
   Abstract »    Full Text »    PDF »
Mechanisms of Nitrosylation and Denitrosylation of Cytoplasmic Glyceraldehyde-3-phosphate Dehydrogenase from Arabidopsis thaliana.
M. Zaffagnini, S. Morisse, M. Bedhomme, C. H. Marchand, M. Festa, N. Rouhier, S. D. Lemaire, and P. Trost (2013)
J. Biol. Chem. 288, 22777-22789
   Abstract »    Full Text »    PDF »
Multilevel Regulation of 2-Cys Peroxiredoxin Reaction Cycle by S-Nitrosylation.
R. Engelman, P. Weisman-Shomer, T. Ziv, J. Xu, E. S. J. Arner, and M. Benhar (2013)
J. Biol. Chem. 288, 11312-11324
   Abstract »    Full Text »    PDF »
Reduction of cardiomyocyte S-nitrosylation by S-nitrosoglutathione reductase protects against sepsis-induced myocardial depression.
P. Y. Sips, T. Irie, L. Zou, S. Shinozaki, M. Sakai, N. Shimizu, R. Nguyen, J. S. Stamler, W. Chao, M. Kaneki, et al. (2013)
Am J Physiol Heart Circ Physiol 304, H1134-H1146
   Abstract »    Full Text »    PDF »
Small changes huge impact: the role of thioredoxin 1 in the regulation of apoptosis by S-nitrosylation.
H. Li, A. Wan, G. Xu, and D. Ye (2013)
Acta Biochim Biophys Sin 45, 153-161
   Abstract »    Full Text »    PDF »
Nitric Oxide Regulates Mitochondrial Fatty Acid Metabolism Through Reversible Protein S-Nitrosylation.
P.-T. Doulias, M. Tenopoulou, J. L. Greene, K. Raju, and H. Ischiropoulos (2013)
Science Signaling 6, rs1
   Abstract »    Full Text »    PDF »
Endogenously produced nitric oxide mitigates sensitivity of melanoma cells to cisplatin.
L. C. Godoy, C. T. M. Anderson, R. Chowdhury, L. J. Trudel, and G. N. Wogan (2012)
PNAS 109, 20373-20378
   Abstract »    Full Text »    PDF »
cGMP-Dependent Activation of Protein Kinase G Precludes Disulfide Activation: Implications for Blood Pressure Control.
J. R. Burgoyne, O. Prysyazhna, O. Rudyk, and P. Eaton (2012)
Hypertension 60, 1301-1308
   Abstract »    Full Text »    PDF »
Nitroglycerin Fails to Lower Blood Pressure in Redox-Dead Cys42Ser PKG1{alpha} Knock-In Mouse.
O. Rudyk, O. Prysyazhna, J. R. Burgoyne, and P. Eaton (2012)
Circulation 126, 287-295
   Abstract »    Full Text »    PDF »
Nitrite is a positive modulator of the Frank-Starling response in the vertebrate heart.
T. Angelone, A. Gattuso, S. Imbrogno, R. Mazza, and B. Tota (2012)
Am J Physiol Regulatory Integrative Comp Physiol 302, R1271-R1281
   Abstract »    Full Text »    PDF »
Thioredoxin-1 Regulates Cellular Heme Insertion by Controlling S-Nitrosation of Glyceraldehyde-3-phosphate Dehydrogenase.
R. Chakravarti and D. J. Stuehr (2012)
J. Biol. Chem. 287, 16179-16186
   Abstract »    Full Text »    PDF »
Physiological Implications of Hydrogen Sulfide: A Whiff Exploration That Blossomed.
R. Wang (2012)
Physiol Rev 92, 791-896
   Abstract »    Full Text »    PDF »
Regulatory Control or Oxidative Damage? Proteomic Approaches to Interrogate the Role of Cysteine Oxidation Status in Biological Processes.
J. M. Held and B. W. Gibson (2012)
Mol. Cell. Proteomics 11, R111.013037
   Abstract »    Full Text »    PDF »
Dynamic denitrosylation via S-nitrosoglutathione reductase regulates cardiovascular function.
F. Beigi, D. R. Gonzalez, K. M. Minhas, Q.-A. Sun, M. W. Foster, S. A. Khan, A. V. Treuer, R. A. Dulce, R. W. Harrison, R. M. Saraiva, et al. (2012)
PNAS 109, 4314-4319
   Abstract »    Full Text »    PDF »
Nitrite-Mediated S-Nitrosylation of Caspase-3 Prevents Hypoxia-Induced Endothelial Barrier Dysfunction.
Y.-C. Lai, K.-T. Pan, G.-F. Chang, C.-H. Hsu, K.-H. Khoo, C.-H. Hung, Y.-J. Jiang, F.-M. Ho, and T.-C. Meng (2011)
Circ. Res. 109, 1375-1386
   Abstract »    Full Text »    PDF »
Site-specific and redox-controlled S-nitrosation of thioredoxin.
K. T. Barglow, C. G. Knutson, J. S. Wishnok, S. R. Tannenbaum, and M. A. Marletta (2011)
PNAS 108, E600-E606
   Abstract »    Full Text »    PDF »
SNO-ing at the Nociceptive Synapse?.
I. Tegeder, R. Scheving, I. Wittig, and G. Geisslinger (2011)
Pharmacol. Rev. 63, 366-389
   Abstract »    Full Text »    PDF »
Thioredoxin Increases Exocytosis by Denitrosylating N-Ethylmaleimide-sensitive Factor.
T. Ito, M. Yamakuchi, and C. J. Lowenstein (2011)
J. Biol. Chem. 286, 11179-11184
   Abstract »    Full Text »    PDF »
Activation of GluR6-containing Kainate Receptors Induces Ubiquitin-dependent Bcl-2 Degradation via Denitrosylation in the Rat Hippocampus after Kainate Treatment.
J. Zhang, H. Yan, Y.-P. Wu, C. Li, and G.-Y. Zhang (2011)
J. Biol. Chem. 286, 7669-7680
   Abstract »    Full Text »    PDF »
Pharmacologically Augmented S-Nitrosylated Hemoglobin Improves Recovery From Murine Subarachnoid Hemorrhage.
H. Sheng, J. D. Reynolds, R. L. Auten, I. T. Demchenko, C. A. Piantadosi, J. S. Stamler, and D. S. Warner (2011)
Stroke 42, 471-476
   Abstract »    Full Text »    PDF »
Protein S-Nitrosylation in Plants: Photorespiratory Metabolism and NO Signaling.
K. J. Gupta (2011)
Science Signaling 4, jc1
   Abstract »    Full Text »    PDF »
Nitrosative modifications of protein and lipid signaling molecules by reactive nitrogen species.
P. J. White, A. Charbonneau, G. J. Cooney, and A. Marette (2010)
Am J Physiol Endocrinol Metab 299, E868-E878
   Abstract »    Full Text »    PDF »
GAPDH regulates cellular heme insertion into inducible nitric oxide synthase.
R. Chakravarti, K. S. Aulak, P. L. Fox, and D. J. Stuehr (2010)
PNAS 107, 18004-18009
   Abstract »    Full Text »    PDF »
Redox Regulatory Mechanism of Transnitrosylation by Thioredoxin.
C. Wu, T. Liu, W. Chen, S.-i. Oka, C. Fu, M. R. Jain, A. M. Parrott, A. T. Baykal, J. Sadoshima, and H. Li (2010)
Mol. Cell. Proteomics 9, 2262-2275
   Abstract »    Full Text »    PDF »
Structural profiling of endogenous S-nitrosocysteine residues reveals unique features that accommodate diverse mechanisms for protein S-nitrosylation.
P.-T. Doulias, J. L. Greene, T. M. Greco, M. Tenopoulou, S. H. Seeholzer, R. L. Dunbrack, and H. Ischiropoulos (2010)
PNAS 107, 16958-16963
   Abstract »    Full Text »    PDF »
S-Nitrosylation Regulates Nuclear Translocation of Chloride Intracellular Channel Protein CLIC4.
M. Malik, A. Shukla, P. Amin, W. Niedelman, J. Lee, K. Jividen, J. M. Phang, J. Ding, K. S. Suh, P. M. G. Curmi, et al. (2010)
J. Biol. Chem. 285, 23818-23828
   Abstract »    Full Text »    PDF »
S-Nitrosylation in Cardiovascular Signaling.
B. Lima, M. T. Forrester, D. T. Hess, and J. S. Stamler (2010)
Circ. Res. 106, 633-646
   Abstract »    Full Text »    PDF »
Protein S-Nitrosylation and Cardioprotection.
J. Sun and E. Murphy (2010)
Circ. Res. 106, 285-296
   Abstract »    Full Text »    PDF »
Pharmacological Impairment of S-Nitrosoglutathione or Thioredoxin Reductases Augments Protein S-Nitrosation in Human Hepatocarcinoma Cells.
Anticancer Res 30, 415-421
   Abstract »    Full Text »    PDF »
Bacterial nitric oxide detoxification prevents host cell S-nitrosothiol formation: a novel mechanism of bacterial pathogenesis.
J. R. Laver, T. M. Stevanin, S. L. Messenger, A. D. Lunn, M. E. Lee, J. W. B. Moir, R. K. Poole, and R. C. Read (2010)
FASEB J 24, 286-295
   Abstract »    Full Text »    PDF »
Thioredoxin-interacting Protein (Txnip) Is a Feedback Regulator of S-Nitrosylation.
M. T. Forrester, D. Seth, A. Hausladen, C. E. Eyler, M. W. Foster, A. Matsumoto, M. Benhar, H. E. Marshall, and J. S. Stamler (2009)
J. Biol. Chem. 284, 36160-36166
   Abstract »    Full Text »    PDF »
Differential effects of reactive nitrogen species on DNA base excision repair initiated by the alkyladenine DNA glycosylase.
L. E. Jones Jr, L. Ying, A. B. Hofseth, E. Jelezcova, R. W. Sobol, S. Ambs, C. C. Harris, M. G. Espey, L. J. Hofseth, and M. D. Wyatt (2009)
Carcinogenesis 30, 2123-2129
   Abstract »    Full Text »    PDF »
H2S Signals Through Protein S-Sulfhydration.
A. K. Mustafa, M. M. Gadalla, N. Sen, S. Kim, W. Mu, S. K. Gazi, R. K. Barrow, G. Yang, R. Wang, and S. H. Snyder (2009)
Science Signaling 2, ra72
   Abstract »    Full Text »    PDF »
Transnitrosylating Nitric Oxide Species Directly Activate Type I Protein Kinase A, Providing a Novel Adenylate Cyclase-independent Cross-talk to {beta}-Adrenergic-like Signaling.
J. R. Burgoyne and P. Eaton (2009)
J. Biol. Chem. 284, 29260-29268
   Abstract »    Full Text »    PDF »
S-nitrosylation of stargazin regulates surface expression of AMPA-glutamate neurotransmitter receptors.
B. Selvakumar, R. L. Huganir, and S. H. Snyder (2009)
PNAS 106, 16440-16445
   Abstract »    Full Text »    PDF »
Kinetic and Cellular Characterization of Novel Inhibitors of S-Nitrosoglutathione Reductase.
P. C. Sanghani, W. I. Davis, S. L. Fears, S.-L. Green, L. Zhai, Y. Tang, E. Martin, N. S. Bryan, and S. P. Sanghani (2009)
J. Biol. Chem. 284, 24354-24362
   Abstract »    Full Text »    PDF »
Estrogen Receptor-{beta} Activation Results in S-Nitrosylation of Proteins Involved in Cardioprotection.
J. Lin, C. Steenbergen, E. Murphy, and J. Sun (2009)
Circulation 120, 245-254
   Abstract »    Full Text »    PDF »
Signaling by Gasotransmitters.
A. K. Mustafa, M. M. Gadalla, and S. H. Snyder (2009)
Science Signaling 2, re2
   Abstract »    Full Text »    PDF »
Redox amplification of apoptosis by caspase-dependent cleavage of glutaredoxin 1 and S-glutathionylation of Fas.
V. Anathy, S. W. Aesif, A. S. Guala, M. Havermans, N. L. Reynaert, Y.-S. Ho, R. C. Budd, and Y. M.W. Janssen-Heininger (2009)
J. Cell Biol. 184, 241-252
   Abstract »    Full Text »    PDF »
Depletion of the ATPase NSF from Golgi membranes with hypo-S-nitrosylation of vasorelevant proteins in endothelial cells exposed to monocrotaline pyrrole.
S. Mukhopadhyay, J. Lee, and P. B. Sehgal (2008)
Am J Physiol Heart Circ Physiol 295, H1943-H1955
   Abstract »    Full Text »    PDF »
Reactive Oxygen Species: Finding the Right Balance.
C. N. Morrell (2008)
Circ. Res. 103, 571-572
   Full Text »    PDF »
Regulation of the Catalytic Activity and Structure of Human Thioredoxin 1 via Oxidation and S-Nitrosylation of Cysteine Residues.
S. I. Hashemy and A. Holmgren (2008)
J. Biol. Chem. 283, 21890-21898
   Abstract »    Full Text »    PDF »
A. Holmgren (2008)
Science 320, 1019-1020
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882