Proteomic analysis of protein tyrosine nitration after ischemia reperfusion injury: mitochondria as the major target.
Endothelial nitric oxide synthase-derived NO and its derivative, peroxynitrite (ONOO(-)), suppresses oxygen consumption by nitration of mitochondrial proteins after reperfusion. However, very few nitrated proteins are identified to date. In this paper, ischemia/reperfusion (I/R) injury was induced in mouse heart by ligation and release of the left anterior descending coronary artery. Western blotting showed that tyrosine nitration was higher in I/R hearts. Nitrated proteins were identified by capillary-liquid chromatography-nanospray tandem mass spectrometry. A total of 23 proteins were identified as being nitrated after I/R and 10 of them were from mitochondria. The nitrated mitochondrial proteins included 4 subunits from the oxidative phosphorylation system (the 24 and the 30 kDa subunits of complex I, the Rieske ISP of complex III, and the alpha subunit of ATP synthase), five enzymes in the matrix, and voltage-dependent anion channel. In purified complex I treated with ONOO(-), 3-NT was identified locating at the residue of Y247 of the 30 kDa subunit and the residues of Y47, Y53 of the 49 kDa subunit. In conclusion, I/R induced protein nitration and mitochondrial proteins were the major targets. Selective nitration of proteins from the oxidative phosphorylation system at the beginning of reperfusion may contribute to the suppression of oxygen consumption.
Liu
Bin
B
Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, and Division of Cardiovascular Medicine, Department of Internal Medicine, College of Medicine, the Ohio State University, Columbus, OH, 43210, USA.
Tewari
Arun K
AK
Zhang
Liwen
L
Green-Church
Kari B
KB
Zweier
Jay L
JL
Chen
Yeong-Renn
YR
He
Guanglong
G
D000818:Animals; D015153:Blotting, Western; D042967:Electron Transport Complex I / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D051379:Mice; D008929:Mitochondria, Heart / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism; D015428:Myocardial Reperfusion Injury / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism*
D024101:Mitochondrial Proteins; D009566:Nitrates; C002744:3-nitrotyrosine; D014443:Tyrosine; D042967:Electron Transport Complex I