Protein glutathionylation: coupling and uncoupling of glutathione to protein thiol groups in lymphocytes under oxidative stress and HIV infection

Ghezzi, Pietro; Romines, Brie; Fratelli, Maddalena; Eberini, Ivano; Gianazza, Elisabetta; Casagrande, Simona; Laragione, Teresa; Mengozzi, Manuela; Herzenberg, Leonore A.; Herzenberg, Leonard A.

doi:10.1016/s0161-5890(01)00114-6

Cited by 89 publications

(51 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Diamide treatment has been shown to also represent a valuable model of redox stress induction (56). Therefore, we tested next whether apoptosis in HtTAM cells was inducible by low concentrations of diamide and could be reversed by induction of MIF.…”

Section: Resultsmentioning

confidence: 99%

The Cytokine Macrophage Migration Inhibitory Factor Reduces Pro-Oxidative Stress-Induced Apoptosis

Nguyen

Lue

Kleemann

et al. 2003

The Journal of Immunology

117

104

View full text Add to dashboard Cite

The cytokine macrophage migration inhibitory factor (MIF) exhibits pro- and anti-inflammatory activities and regulates cell proliferation and survival. We investigated the effects of MIF on apoptosis. As MIF exhibits oxidoreductase activity and participates in regulating oxidative cell stress, we studied whether MIF could affect oxidative stress-induced apoptosis. We demonstrated that MIF exhibits antiapoptotic activity in various settings. MIF suppressed camptothecin-induced apoptosis in HeLa and Kym cells and HL-60 promyeloblasts. Both exogenous MIF and endogenous MIF, induced following overexpression through tetracycline (tet) gene induction, led to significant suppression of apoptosis. Apoptosis reduction by MIF was also observed in T cells. A role for MIF in redox stress-induced apoptosis was addressed by comparing the effects of rMIF with those of the oxidoreductase mutant C60SMIF. Endogenous overexpression of C60SMIF was similar to that of MIF, but C60SMIF did not suppress apoptosis. Exogenous rC60SMIF inhibited apoptosis. A role for MIF in oxidative stress-induced apoptosis was directly studied in HL-60 leukocytes and tet-regulated HeLa cells following thiol starvation or diamide treatment. MIF protected these cells from redox stress-induced apoptosis and enhanced cellular glutathione levels. As overexpressed C60SMIF did not protect tet-regulated HeLa cells from thiol starvation-induced apoptosis, it seems that the redox motif of MIF is important for this function. Finally, overexpression of MIF inhibited phosphorylation of endogenous c-Jun induced by thiol starvation, indicating that MIF-based suppression of apoptosis is mediated through modulation of c-Jun N-terminal kinase activity. Our findings show that MIF has potent antiapoptotic activities and suggest that MIF is a modulator of pro-oxidative stress-induced apoptosis.

show abstract

Section: Resultsmentioning

confidence: 99%

The Cytokine Macrophage Migration Inhibitory Factor Reduces Pro-Oxidative Stress-Induced Apoptosis

Nguyen

Lue

Kleemann

et al. 2003

The Journal of Immunology

117

104

View full text Add to dashboard Cite

show abstract

“…Data were compared with the theoretical molecular weight of the rhTrx tryptic peptides, taking into consideration the possible glutathionylated Cys. Previous studies have shown that under these experimental conditions, diamide induces marked 35 S incorporation into proteins (19).…”

Section: Titration Of Free -Sh Groups With 5-5-dithiobis-2-nitrobenzomentioning

confidence: 84%

Glutathionylation of human thioredoxin: A possible crosstalk between the glutathione and thioredoxin systems

Casagrande

Bonetto

Fratelli

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

323

203

View full text Add to dashboard Cite

To identify proteins undergoing glutathionylation (formation of protein-glutathione mixed disulfides) in human T cell blasts, we radiolabeled the glutathione pool with 35 S, exposed cells to the oxidant diamide, and analyzed cellular proteins by two-dimensional electrophoresis. One of the proteins undergoing glutathionylation was identified by molecular weight, isoelectric point, and immunoblotting as thioredoxin (Trx). Incubation of recombinant human Trx with glutathione disulfide or S-nitrosoglutathione led to the formation of glutathionylated Trx, identified by matrixassisted laser desorption ionization-time-of-flight mass spectrometry. The glutathionylation site was identified as Cys-72. Glutathionylation of rhTrx abolished its enzymatic activity as insulin disulfide reductase in the presence of NADPH and Trx reductase. Activity was, however, regained with sigmoidal kinetics, indicating a process of autoactivation due to the ability of Trx to deglutathionylate itself. These data suggest that the intracellular glutathione͞glutathione disulfide ratio, an indicator of the redox state of the cell, can regulate Trx functions reversibly through thiol-disulfide exchange reactions. T hioredoxin (Trx; ref. 1), a ubiquitous redox protein, is an essential cofactor electron donor for ribonucleotide reductase, but also has many other cellular functions, including regulation of transcription factors, apoptosis, and antioxidant activity and can act exogenously as a redox active growth factor (1, 2). The catalytic activity of Trx resides in its active site where the two redox active Cyss (Cys-31 and Cys-34 in human Trx) undergo reversible oxidation͞reduction. In addition to the conserved Cys residues in the active site, three additional structural Cys residues (Cys-61, Cys-68, and Cys-72) are present in the structure of human Trx.The present paper describes the experiments that led us to the conclusion that Trx can undergo glutathionylation in T cells exposed to oxidative stress. Protein glutathionylation, the formation of a disulfide between a Cys in a protein and the Cys in the tripeptide glutathione (GSH), is a modification that can be induced in cells by oxidative stress. Glutathionylation can occur by direct oxidation of a protein and GSH, by a thiol-disulfide exchange between a protein Cys and oxidized glutathione (GSSG), and also with the intermediacy of S-nitrosoglutathione (GSNO; refs. 3 and 4). Glutathionylation of proteins is reversible, as those proteins can be reduced by glutaredoxins (5, 6), and the process serves to regulate protein functions by the redox state of the cell (i.e., by the GSSG͞GSH ratio; refs. 7-9).To demonstrate the glutathionylation of Trx, we first labeled the intracellular GSH pool of the T-cell blasts with [ 35 S]Cys and analyzed the cell lysate by two dimensional electrophoresis under nonreducing conditions. A spot was found in the autoradiographic protein map with IP͞M r corresponding to those of Trx. In the second part of the study, we incubated recombinant human Trx in the presence or ...

show abstract

“…Glutathione (GSH) is an abundant anti-oxidant in cells and plays a critical role in protection from oxidative damage (6). Protein S-glutathionylation, the disulfide coupling of a GSH moiety to Cys residues, is the prevalent S-thiolation reaction in biological systems (7)(8), regulating numerous physiological processes (9). This modification may be driven by oxidative/ nitrosative stress in the presence of endogenous GSH but can also persist under basal conditions and in reducing environments (10 -11).…”

mentioning

confidence: 99%

S-Glutathionylation Regulates Inflammatory Activities of S100A9

Lim

Raftery

Goyette

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Reactive oxygen species generated by activated neutrophils can cause oxidative stress and tissue damage. S100A8 (A8) and S100A9 (A9), abundant in neutrophil cytoplasm, are exquisitely sensitive to oxidation, which may alter their functions. Murine A8 is a neutrophil chemoattractant, but it suppresses leukocyte transmigration in the microcirculation when S-nitrosylated. Glutathione (GSH) modulates intracellular redox, and S-glutathionylation can protect susceptible proteins from oxidative damage and regulate function. We characterized S-glutathionylation of A9; GSSG and GSNO generated S-glutathionylated A8 (A8-SSG) and A9 (A9-SSG) in vitro, whereas only A9-SSG was detected in cytosol of neutrophils activated with phorbol myristate acetate (PMA) but not with fMLP or opsonized zymosan. S-Glutathionylation exposed more hydrophobic regions in Zn 2؉ -bound A9 but did not alter Zn 2؉ binding affinity. A9-SSG had reduced capacity to form heterocomplexes with A8, but the arachidonic acid binding capacities of A8/A9 and A8/A9-SSG were similar. A9 and A8/A9 bind endothelial cells; S-glutathionylation reduced binding. We found little effect of A9 or A9-SSG on neutrophil CD11b/CD18 expression or neutrophil adhesion to endothelial cells. However, A9, A9-SSG and A8/A9 promoted neutrophil adhesion to fibronectin but, in the presence of A8, A9-mediated adhesion was abrogated by glutathionylation. S-Glutathionylation of A9 may protect its oxidation to higher oligomers and reduce neutrophil binding to the extracellular matrix. This may regulate the magnitude of neutrophil migration in the extravasculature, and together with the functional changes we reported for S-nitrosylated A8, particular oxidative modifications of these proteins may limit tissue damage in acute inflammation.

show abstract

Protein glutathionylation: coupling and uncoupling of glutathione to protein thiol groups in lymphocytes under oxidative stress and HIV infection

Cited by 89 publications

References 32 publications

The Cytokine Macrophage Migration Inhibitory Factor Reduces Pro-Oxidative Stress-Induced Apoptosis

The Cytokine Macrophage Migration Inhibitory Factor Reduces Pro-Oxidative Stress-Induced Apoptosis

Glutathionylation of human thioredoxin: A possible crosstalk between the glutathione and thioredoxin systems

S-Glutathionylation Regulates Inflammatory Activities of S100A9

Contact Info

Product

Resources

About