Preconditioning results in S-nitrosylation of proteins involved in regulation of mitochondrial energetics and calcium transport

Junhui Sun, Meghan Morgan, Rong Fong Shen, Charles Steenbergen, Elizabeth Murphy

Research output: Contribution to journalArticlepeer-review

300 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)1155-1163
Number of pages9
JournalCirculation research
Issue number11
StatePublished - Nov 2007


  • Cardioprotection
  • Preconditioning
  • S-nitrosylation

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine


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