Mechanism of superoxide dismutase/H(2)O(2)-mediated nitric oxide release from S-nitrosoglutathione--role of glutamate.
S-Nitrosoglutathione (GSNO), a physiologically relevant nitric oxide ((*)NO) donor, exhibits antioxidant, anti-ischemic, and antiplatelet properties. The exact mechanism of (*)NO release from GSNO in biological systems has not been determined. Both copper ions and copper-containing enzymes have been shown to catalyze (*)NO release from GSNO. In this study we observed that copper-zinc superoxide dismutase (Cu,ZnSOD) in the presence of H(2)O(2) caused a rapid decomposition of GSNO, forming oxidized glutathione (GSSG) and (*)NO. The cupric ions (Cu(2+)) released from Cu,ZnSOD were bound to the glutamate moiety of GSNO, yielding a 2:1 (GSNO)(2)Cu(2+) complex. Strong chelators of cupric ions, such as histidine and diethylenetriaminepentaacetic acid, inhibited the formation of (GSNO)(2)Cu(2+) complex, GSSG, and (*)NO. GSSG alone inhibited Cu(2+)-induced decomposition of GSNO. This effect is attributed to complexation of copper by GSSG. We conclude that binding of copper to GSNO is obligatory for (*)NO release from GSNO; however, the rate of this reaction was considerably slowed due to binding of Cu(2+) by GSSG. The glutamate moiety in GSNO and GSSG controls copper-catalyzed (*)NO release from GSNO. Cu,ZnSOD and H(2)O(2) enhanced peroxidation of unsaturated lipid that was inhibited by GSNO. The antioxidant function of GSNO is related to the sequestering of copper by GSNO and its ability to slowly release (*)NO. Implications of these findings are discussed in relation to GSNO-induced cardioprotection and to neuropathological processes.
Singh
R J
RJ
Biophysics Research Institute, Medical College of Wisconsin, Milwaukee, Wisconsin, 53226, USA.
Hogg
N
N
Goss
S P
SP
Antholine
W E
WE
Kalyanaraman
B
B
D000818:Animals; D000975:Antioxidants / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D002417:Cattle; D003300:Copper / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D018698:Glutamic Acid / Q000378:metabolism*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000378:metabolism*; D066298:In Vitro Techniques; D015227:Lipid Peroxidation / Q000187:drug effects; D008081:Liposomes; D009569:Nitric Oxide / Q000378:metabolism*; D009603:Nitroso Compounds / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D026422:S-Nitrosoglutathione; D013482:Superoxide Dismutase / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism
D000975:Antioxidants; D008081:Liposomes; D009603:Nitroso Compounds; D009569:Nitric Oxide; D018698:Glutamic Acid; D026422:S-Nitrosoglutathione; D003300:Copper; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D005978:Glutathione; D015032:Zinc