Preconditioning Results in S-Nitrosylation of Proteins Involved in Regulation of Mitochondrial Energetics and Calcium Transport

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 2...

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Published in:	Circulation research 2007-11, Vol.101 (11), p.1155-1163
Main Authors:	Sun, Junhui, Morgan, Meghan, Shen, Rong-Fong, Steenbergen, Charles, Murphy, Elizabeth
Format:	Article
Language:	eng
Subjects:	Animals Biological and medical sciences Biological Transport Calcium - metabolism Cardiotonic Agents Energy Metabolism Fundamental and applied biological sciences. Psychology In Vitro Techniques Ischemic Preconditioning, Myocardial - methods Mice Mice, Inbred C57BL Mitochondria - metabolism Myocardial Reperfusion Injury - prevention & control Nitrogen Oxides - metabolism Perfusion Proteins - metabolism Ventricular Dysfunction, Left Vertebrates: cardiovascular system
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Summary:	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 Ca channel α1 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 Ca-ATPase, α-ketoglutarate dehydrogenase, and the mitochondrial F1-ATPase α1 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.
ISSN:	0009-7330 1524-4571