Selective nitration of mitochondrial complex I by peroxynitrite: involvement in mitochondria dysfunction and cell death of dopaminergic SH-SY5Y cells.
3-Nitrotyrosine (3-NT) is a specific marker of protein nitration by peroxynitrite (ONOO-) produced from nitric oxide and superoxide. Increase in 3-NT containing protein (3-NT protein) was reported in brains from patients with some neurodegenerative disorders and aging. In this paper, intracellular localization of 3-NT protein was examined in dopaminergic SH-SY5Y cells using the selective antibody against protein-bound 3-NT. 3-NT protein was detected in plasma membrane/nucleus and mitochondria fractions, and interestingly in polypeptide composition of mitochondrial complex I. ONOO--generating SIN-1 induced apoptotic cell death with concomitant increase in 3-NT protein and reduction in mitochondrial ATP synthesis. In addition, an inhibitor of proteasomes, carbobenzoxy-L-isoleucyl-gamma-t-butyl-L-glutamyl-L-alanyl-L-leucinal, enhanced the effects of ONOO-. These results suggest that ONOO- may induce mitochondrial dysfunction and cell death in neurons through nitration of mitochondrial complex I subunits.
Yamamoto
T
T
Research Centre of Health, Physical Fitness and Sports, Nagoya University, Chikusa-ku, Nagoya, Japan.
Maruyama
W
W
Kato
Y
Y
Yi
H
H
Shamoto-Nagai
M
M
Tanaka
M
M
Sato
Y
Y
Naoi
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis; D016923:Cell Death; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism; D042967:Electron Transport Complex I; D006801:Humans; D008928:Mitochondria / Q000502:physiology*; D008981:Molsidomine / Q000031:analogs & derivatives* / Q000494:pharmacology; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D030421:Peroxynitrous Acid / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D015853:Cysteine Proteinase Inhibitors; D020030:Nitric Oxide Donors; D009842:Oligopeptides; C090535:benzyloxycarbonyl-isoleucyl-glutamyl(O-tert-butyl)-alanyl-leucinal; D030421:Peroxynitrous Acid; C002744:3-nitrotyrosine; D014443:Tyrosine; C002385:linsidomine; D000255:Adenosine Triphosphate; D008981:Molsidomine; D009247:NADH, NADPH Oxidoreductases; D042967:Electron Transport Complex I; D004298:Dopamine