Nitric oxide-induced S-glutathionylation and inactivation of glyceraldehyde-3-phosphate dehydrogenase.
S-Nitrosylation of protein thiol groups by nitric oxide (NO) is a widely recognized protein modification. In this study we show that nitrosonium tetrafluoroborate (BF4NO), a NO+ donor, modified the thiol groups of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by S-nitrosylation and caused enzyme inhibition. The resultant protein-S-nitrosothiol was found to be unstable and to decompose spontaneously, thereby restoring enzyme activity. In contrast, the NO-releasing compound S-nitrosoglutathione (GSNO) promoted S-glutathionylation of a thiol group of GAPDH both in vitro and under cellular conditions. The GSH-mixed protein disulfide formed led to a permanent enzyme inhibition, but upon dithiothreitol addition a functional active GAPDH was recovered. This S-glutathionylation is specific for GSNO because GSH itself was unable to produce protein-mixed disulfides. During cellular nitrosative stress, the production of intracellular GSNO might channel signaling responses to form protein-mixed disulfide that can regulate intracellular function.
Mohr
S
S
Molecular Cardiovascular Research Center, Case Western Reserve University School of Medicine and the University Hospitals of Cleveland, Cleveland, Ohio 44106-4958, USA.
Hallak
H
H
de Boitte
A
A
Lapetina
E G
EG
Brüne
B
B
D000818:Animals; D001881:Borates / Q000494:pharmacology; D002417:Cattle; D004730:Endothelium, Vascular / Q000378:metabolism; D005033:Ethylmaleimide / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D009569:Nitric Oxide / Q000494:pharmacology*; D009603:Nitroso Compounds / Q000494:pharmacology; D026422:S-Nitrosoglutathione; D013438:Sulfhydryl Compounds / Q000378:metabolism
D001881:Borates; D009603:Nitroso Compounds; D013438:Sulfhydryl Compounds; C088437:nitrosonium tetrafluoroborate; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005978:Glutathione; D005033:Ethylmaleimide