Persistent S-nitrosation of complex I and other mitochondrial membrane proteins by S-nitrosothiols but not nitric oxide or peroxynitrite: implications for the interaction of nitric oxide with mitochondria.

TitlePersistent S-nitrosation of complex I and other mitochondrial membrane proteins by S-nitrosothiols but not nitric oxide or peroxynitrite: implications for the interaction of nitric oxide with mitochondria.
Publication TypeJournal Article
Year of Publication2006
AuthorsDahm, CC, Moore, K, Murphy, MP
JournalJ Biol Chem
Volume281
Issue15
Pagination10056-65
Date Published2006 Apr 14
ISSN0021-9258
KeywordsAnimals, Disulfides, Dose-Response Relationship, Drug, Electrophoresis, Polyacrylamide Gel, Glutathione, Immunoblotting, Intracellular Membranes, Liver, Mitochondria, Mitochondria, Liver, Models, Chemical, Nitric Oxide, Nitrosation, Nitroso Compounds, Oxygen Consumption, Parkinson Disease, Peroxynitrous Acid, Rats, S-Nitroso-N-Acetylpenicillamine, S-Nitrosothiols, Sepsis, Sulfhydryl Compounds, Superoxides, Time Factors
Abstract

S-nitrosation of mitochondrial proteins has been proposed to contribute to the pathophysiological interactions of nitric oxide (NO) and its derivatives with mitochondria but has not been shown directly. Furthermore, little is known about the mechanism of formation or the fate of these putative S-nitrosothiols. Here we have determined whether mitochondrial membrane protein thiols can be S-nitrosated on exposure to free NO from 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene (DETA-NONOate) by interaction with S-nitrosoglutathione or S-nitroso-N-acetylpenicillamine (SNAP) and by the NO derivative peroxynitrite. S-Nitrosation of protein thiols was measured directly by chemiluminescence detection. S-Nitrosoglutathione and S-nitroso-N-acetylpenicillamine led to extensive protein thiol oxidation, with about 30% of the modified protein thiols persistently S-nitrosated. In contrast, there was no protein thiol oxidation or S-nitrosation on exposure to 3,3-bis (aminoethyl)-1-hydroxy-2-oxo-1-triazene. Peroxynitrite extensively oxidized protein thiols but produced negligible amounts of S-nitrosothiols. Therefore, mitochondrial membrane protein thiols are S-nitrosated by preformed S-nitrosothiols but not by NO or by peroxynitrite. These S-nitrosated protein thiols were readily reduced by glutathione, so S-nitrosation will only persist when the mitochondrial glutathione pool is oxidized. Respiratory chain complex I was S-nitrosated by S-nitrosothiols, consistent with it being an important target for S-nitrosation during nitrosative stress. The S-nitrosation of complex I correlated with a significant loss of activity that was reversed by thiol reductants. S-Nitrosation was also associated with increased superoxide production from complex I. These findings point to a significant role for complex I S-nitrosation and consequent dysfunction during nitrosative stress in disorders such as Parkinson disease and sepsis.

DOI10.1074/jbc.M512203200
Alternate JournalJ. Biol. Chem.
Citation Key10.1074/jbc.M512203200
PubMed ID16481325
Grant ListMC_U105663142 / / Medical Research Council / United Kingdom