TY - JOUR
T1 - Protein glutathionylation
T2 - Coupling and uncoupling of glutathione to protein thiol groups in lymphocytes under oxidative stress and HIV infection
AU - Ghezzi, Pietro
AU - Romines, Brie
AU - Fratelli, Maddalena
AU - Eberini, Ivano
AU - Gianazza, Elisabetta
AU - Casagrande, Simona
AU - Laragione, Teresa
AU - Mengozzi, Manuela
AU - Herzenberg, Leonore A.
AU - Herzenberg, Leonard A.
PY - 2002
Y1 - 2002
N2 - We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein cysteinyl residues (glutathionylation) in T cell blasts. Treating the cells with the oxidant diamide induces thiolation of a series of proteins that can be detected by 2D electrophoresis when 35S-cysteine is used to label the intracellular GSH pool. This thiolation is reversible, proteins are rapidly dethiolated and GSH is released from proteins once the oxidants are washed and the cells are allowed to recover. Dethiolation is dependent on the availability of GSH and thiols, since it is inhibited by GSH-depleting agents and improved by N-acetyl-L-cysteine (NAC). The capacity of these agents to reverse glutathionylation is diminished in T cell blasts infected in vitro with HIV, which is known to cause oxidative stress. Consistent with these findings, the activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an enzyme known to be inhibited by glutathionylation, is inhibited in diamide-treated cells and recovers rapidly when cells are allowed to dethiolate. Further, GAPDH activity is diminished by GSH-depleting agents and augmented by NAC. Thus, reversible glutathionylation of proteins can rapidly shift the activity of a key metabolic enzyme and thereby result in dramatic, reversible changes in cellular metabolism.
AB - We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein cysteinyl residues (glutathionylation) in T cell blasts. Treating the cells with the oxidant diamide induces thiolation of a series of proteins that can be detected by 2D electrophoresis when 35S-cysteine is used to label the intracellular GSH pool. This thiolation is reversible, proteins are rapidly dethiolated and GSH is released from proteins once the oxidants are washed and the cells are allowed to recover. Dethiolation is dependent on the availability of GSH and thiols, since it is inhibited by GSH-depleting agents and improved by N-acetyl-L-cysteine (NAC). The capacity of these agents to reverse glutathionylation is diminished in T cell blasts infected in vitro with HIV, which is known to cause oxidative stress. Consistent with these findings, the activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an enzyme known to be inhibited by glutathionylation, is inhibited in diamide-treated cells and recovers rapidly when cells are allowed to dethiolate. Further, GAPDH activity is diminished by GSH-depleting agents and augmented by NAC. Thus, reversible glutathionylation of proteins can rapidly shift the activity of a key metabolic enzyme and thereby result in dramatic, reversible changes in cellular metabolism.
KW - Glutathione
KW - Lymphocytes
KW - Oxidative stress
KW - Protein glutathionylation
KW - Thiols
UR - http://www.scopus.com/inward/record.url?scp=18244385536&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=18244385536&partnerID=8YFLogxK
U2 - 10.1016/S0161-5890(01)00114-6
DO - 10.1016/S0161-5890(01)00114-6
M3 - Article
C2 - 11841837
AN - SCOPUS:18244385536
VL - 38
SP - 773
EP - 780
JO - Molecular Immunology
JF - Molecular Immunology
SN - 0161-5890
IS - 10
ER -