title: Preconditioning results in S-nitrosylation of proteins involved in regulation of mitochondrial energetics and calcium transport.
abstract: Nitric oxide has been shown to be an important signaling messenger in ischemic preconditioning (IPC). Accordingly, we investigated whether protein S-nitrosylation occurs in IPC hearts and whether S-nitrosoglutathione (GSNO) elicits similar effects on S-nitrosylation and cardioprotection. Preceding 20 minutes of no-flow ischemia and reperfusion, hearts from C57BL/6J mice were perfused in the Langendorff mode and subjected to the following conditions: (1) control perfusion; (2) IPC; or (3) 0.1 mmol/L GSNO treatment. Compared with control, IPC and GSNO significantly improved postischemic recovery of left ventricular developed pressure and reduced infarct size. IPC and GSNO both significantly increased S-nitrosothiol contents and S-nitrosylation levels of the L-type Ca2+ channel alpha1 subunit in heart membrane fractions. We identified several candidate S-nitrosylated proteins by proteomic analysis following the biotin switch method, including the cardiac sarcoplasmic reticulum Ca2+-ATPase, alpha-ketoglutarate dehydrogenase, and the mitochondrial F1-ATPase alpha1 subunit. The activities of these enzymes were altered in a concentration-dependent manner by GSNO treatment. We further developed a 2D DyLight fluorescence difference gel electrophoresis proteomic method that used DyLight fluors and a modified biotin switch method to identify S-nitrosylated proteins. IPC and GSNO produced a similar pattern of S-nitrosylation modification and cardiac protection against ischemia/reperfusion injury, suggesting that protein S-nitrosylation may play an important cardioprotective role in heart.
authors.affiliation: NHLBI, NIH, Vascular Medicine Branch, Bethesda, MD 20892, USA.
authors.forename: Junhui
authors.initials: J
authors.lastname: Sun
authors.forename: Meghan
authors.initials: M
authors.lastname: Morgan
authors.forename: Rong-Fong
authors.initials: RF
authors.lastname: Shen
authors.forename: Charles
authors.initials: C
authors.lastname: Steenbergen
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Murphy
mesh_terms: D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D002316:Cardiotonic Agents; D004734:Energy Metabolism; D066298:In Vitro Techniques; D019157:Ischemic Preconditioning, Myocardial / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D009589:Nitrogen Oxides / Q000378:metabolism*; D010477:Perfusion; D011506:Proteins / Q000378:metabolism*; D018487:Ventricular Dysfunction, Left
chemical_list: D002316:Cardiotonic Agents; D009589:Nitrogen Oxides; D011506:Proteins; D002118:Calcium