The Journal of Immunology, Vol 156, Issue 5 1973-1980, Copyright © 1996 by American Association of Immunologists

ARTICLES

Protein S-thiolation and dethiolation during the respiratory burst in human monocytes. A reversible post-translational modification with potential for buffering the effects of oxidant stress

T Seres, V Ravichandran, T Moriguchi, K Rokutan, JA Thomas and RB Johnston Jr
Department of Pediatrics (Immunology), Yale University School of Medicine, New Haven, CT 06520, USA.

Stimulation of the respiratory burst in mouse macrophages or human neutrophils results in the formation of disulfide bonds between low m.w. thiols and sulfhydryl groups on specific cytosolic proteins (S- thiolation). S-thiolation is reversible in certain chemical systems. The aim of the present study was to analyze the dynamic nature of this process in human monocytes under physiologic conditions. We report here that the extent of S-thiolation and the rate of respiratory burst stimulated by opsonized zymosan or phorbol diester increased for 10 to 20 min and then declined (dethiolation) in close association. Individual proteins underwent S-thiolation and dethiolation at different rates. H2O--appeared particularly effective in mediating S- thiolation, based on inhibition of S-thiolation by added catalase and accentuation by azide, which inhibits cellular catalase. S-thiolation did not occur in stimulated monocytes from patients with chronic granulomatous disease. The addition of H2O2 to monocytes or lymphocytes induced rapid S-thiolation (1 to 3 min); a subsequent dethiolation returned most of the proteins to baseline by 15 to 30 min. At 0 degrees C and after addition of 1,3-bis-(2-chloroethyl)-1-nitrosourea, there was effective S-thiolation on exposure to H2O2, but dethiolation was inhibited, suggesting a possible role for glutathione (GSH)/thioredoxin reductase systems in this process. GSH was determined to be the most abundant low m.w. thiol bound to S-thiolated proteins, but gamma- glutamylcysteine and cysteine were also bound. The time of maximal reduction in cytosolic GSH during the respiratory burst (10 min) coincided with the time at which protein-bound GSH was highest. S- thiolation-dethiolation represents a reversible post-translational modification that could protect cellular proteins from irreversible oxidative damage.

This article has been cited by other articles:

				X.-H. Gao, M. Bedhomme, D. Veyel, M. Zaffagnini, and S. D. Lemaire Methods for Analysis of Protein Glutathionylation and their Application to Photosynthetic Organisms Mol Plant, March 1, 2009; 2(2): 218 - 235. [Abstract] [Full Text] [PDF]

				R. E. Hansen, D. Roth, and J. R. Winther Quantifying the global cellular thiol-disulfide status PNAS, January 13, 2009; 106(2): 422 - 427. [Abstract] [Full Text] [PDF]

				M. B. West, B. G. Hill, Y.-T. Xuan, and A. Bhatnagar Protein glutathiolation by nitric oxide: an intracellular mechanism regulating redox protein modification FASEB J, August 1, 2006; 20(10): 1715 - 1717. [Abstract] [Full Text] [PDF]

				S. Qanungo, M. Wang, and A.-L. Nieminen N-Acetyl-L-cysteine Enhances Apoptosis through Inhibition of Nuclear Factor-{kappa}B in Hypoxic Murine Embryonic Fibroblasts J. Biol. Chem., November 26, 2004; 279(48): 50455 - 50464. [Abstract] [Full Text] [PDF]

				J. F. Caplan, N. R. Filipenko, S. L. Fitzpatrick, and D. M. Waisman Regulation of Annexin A2 by Reversible Glutathionylation J. Biol. Chem., February 27, 2004; 279(9): 7740 - 7750. [Abstract] [Full Text] [PDF]

				R. E. Laliberte, D. G. Perregaux, L. R. Hoth, P. J. Rosner, C. K. Jordan, K. M. Peese, James. F. Eggler, M. A. Dombroski, K. F. Geoghegan, and C. A. Gabel Glutathione S-Transferase Omega 1-1 Is a Target of Cytokine Release Inhibitory Drugs and May Be Responsible for Their Effect on Interleukin-1beta Posttranslational Processing J. Biol. Chem., May 2, 2003; 278(19): 16567 - 16578. [Abstract] [Full Text] [PDF]

				K. C. Das and C. W. White Redox systems of the cell: Possible links and implications PNAS, July 23, 2002; 99(15): 9617 - 9618. [Full Text] [PDF]

				D. Shenton, G. Perrone, K. A. Quinn, I. W. Dawes, and C. M. Grant Regulation of Protein S-Thiolation by Glutaredoxin 5 in the Yeast Saccharomyces cerevisiae J. Biol. Chem., May 3, 2002; 277(19): 16853 - 16859. [Abstract] [Full Text] [PDF]

				S. P. Hehner, R. Breitkreutz, G. Shubinsky, H. Unsoeld, K. Schulze-Osthoff, M. L. Schmitz, and W. Droge Enhancement of T Cell Receptor Signaling by a Mild Oxidative Shift in the Intracellular Thiol Pool J. Immunol., October 15, 2000; 165(8): 4319 - 4328. [Abstract] [Full Text] [PDF]

				T. Seres, R. G. Knickelbein, J. B. Warshaw, and R. B. Johnston Jr. The Phagocytosis-Associated Respiratory Burst in Human Monocytes Is Associated with Increased Uptake of Glutathione J. Immunol., September 15, 2000; 165(6): 3333 - 3340. [Abstract] [Full Text] [PDF]

				E Maellaro, S Dominici, B Del Bello, M. Valentini, L Pieri, P Perego, R Supino, F Zunino, E Lorenzini, A Paolicchi, et al. Membrane gamma-glutamyl transpeptidase activity of melanoma cells: effects on cellular H(2)O(2) production, cell surface protein thiol oxidation and NF-kappa B activation status J. Cell Sci., January 8, 2000; 113(15): 2671 - 2678. [Abstract] [PDF]

				C. Elbim, S. Pillet, M. H. Prevost, A. Preira, P. M. Girard, N. Rogine, H. Matusani, J. Hakim, N. Israel, and M. A. Gougerot-Pocidalo Redox and Activation Status of Monocytes from Human Immunodeficiency Virus-Infected Patients: Relationship with Viral Load J. Virol., June 1, 1999; 73(6): 4561 - 4566. [Abstract] [Full Text]

				C. M. Grant, K. A. Quinn, and I. W. Dawes Differential Protein S-Thiolation of Glyceraldehyde-3-Phosphate Dehydrogenase Isoenzymes Influences Sensitivity to Oxidative Stress Mol. Cell. Biol., April 1, 1999; 19(4): 2650 - 2656. [Abstract] [Full Text] [PDF]

				E. Schmid, J. E. Benna, D. Galter, G. Klein, and W. Dröge Redox priming of the insulin receptor ß-chain associated with altered tyrosine kinase activity and insulin responsiveness in the absence of tyrosine autophosphorylation FASEB J, July 1, 1998; 12(10): 863 - 870. [Abstract] [Full Text]

				P. G. Board, M. Coggan, G. Chelvanayagam, S. Easteal, L. S. Jermiin, G. K. Schulte, D. E. Danley, L. R. Hoth, M. C. Griffor, A. V. Kamath, et al. Identification, Characterization, and Crystal Structure of the Omega Class Glutathione Transferases J. Biol. Chem., August 4, 2000; 275(32): 24798 - 24806. [Abstract] [Full Text] [PDF]

				C. A. Chrestensen, D. W. Starke, and J. J. Mieyal Acute Cadmium Exposure Inactivates Thioltransferase (Glutaredoxin), Inhibits Intracellular Reduction of Protein-glutathionyl-mixed Disulfides, and Initiates Apoptosis J. Biol. Chem., August 18, 2000; 275(34): 26556 - 26565. [Abstract] [Full Text] [PDF]