Protein kinase C-ε coimmunoprecipitates with cytochrome oxidase subunit IV and is associated with improved cytochrome-c oxidase activity and cardioprotection

Dehuang Guo, Tiffany Nguyen, Mourad Ogbi, Huda Tawfik, Guochun Ma, Qilin Yu, Robert W. Caldwell, John A. Johnson

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

We have utilized an in situ rat coronary ligation model to establish a PKC-ε cytochrome oxidase subunit IV (COIV) coimmunoprecipitation in myocardium exposed to ischemic preconditioning (PC). Ischemia-reperfusion (I/R) damage and PC protection were confirmed using tetrazolium-based staining methods and serum levels of cardiac troponin I. Homogenates prepared from the regions at risk (RAR) and not at risk (RNAR) for I/R injury were fractionated into cell-soluble (S), 600 g low-speed centrifugation (L), Percoll/Optiprep density gradient-purified mitochondrial (M), and 100,000 g particulate (P) fractions. COIV immunoreactivity and cytochrome-c oxidase activity measurements estimated the percentages of cellular mitochondria in S, L, M, and P fractions to be 0, 55, 29, and 16%, respectively. We observed 18, 3, and 3% of PKC-δ, -ε, and -ζ isozymes in the M fraction under basal conditions. Following PC, we observed a 61% increase in PKC-ε levels in the RAR M fraction compared with the RNAR M fraction. In RAR mitochondria, we also observed a 2.8-fold increase in PKC-ε serine 729 phosphoimmunoreactivity (autophosphorylation), indicating the presence of activated PKC-ε in mitochondria following PC. PC administered before prolonged I/R induced a 1.9-fold increase in the coimmunoprecipitation of COIV, with anti-PKC-ε antisera and a twofold enhancement of cytochrome-c oxidase activity. Our results suggest that PKC-ε may interact with COIV as a component of the cardioprotection in PC. Induction of this interaction may provide a novel therapeutic target for protecting the heart from I/R damage.

Original languageEnglish (US)
Pages (from-to)H2219-H2230
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume293
Issue number4
DOIs
StatePublished - Oct 1 2007

Keywords

  • Cardiac
  • Coronary ligation
  • Ischemia-reperfusion
  • Mitochondria
  • Oxidative phosphorylation
  • Protein kinase C

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

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