Troponin I degradation and covalent complex formation accompanies myocardial ischemia/reperfusion injury

Jason L. McDonough, D. Kent Arrell, Jennifer E. Van Eyk

Research output: Contribution to journalArticle

Abstract

Selective troponin I (TnI) modification has been demonstrated to be in part responsible for the contractile dysfunction observed with myocardial ischemia/reperfusion injury. We have isolated and characterized modified TnI products in isolated rat hearts after 0, 15, or 60 minutes of ischemia followed by 45 minutes of reperfusion using affinity chromatography with cardiac troponin C (TnC) and an anti-TnI antibody, immunological mapping, reversed-phase high-performance liquid chromatography, and mass spectrometry. Rat cardiac TnI becomes progressively degraded from 210 amino acid residues to residues 1-193, 63-193, and 73-193 with increased severity of injury. Degradation is accompanied by formation of covalent complexes between TnI 1- 193 and, respectively, TnC residues 1-94 and troponin T (TnT) residues 191- 298. The covalent complexes are likely a result of isopeptide bond formation between lysine 193 of TnI and glutamine 191 of TnT by the cross-linking enzyme transglutaminase. With severe ischemia, cellular necrosis results in specific release of TnI 1-193 into the reperfusion effluent and TnT degradation in the myocardium (25-, 27-, and 33-kDa products). Two- dimensional electrophoresis demonstrated that phosphorylation of TnI prevents ischemia-induced degradation. This study characterized the modified TnI products in isolated rat hearts reperfused after a brief or severe period of ischemia, revealing the progressive nature of TnI degradation, changes in phosphorylation, and covalent complexes with ischemia/reperfusion injury. Finally, we propose a model for ischemia/reperfusion injury in which the extent of proteolytic and transglutaminase activities ultimately determines whether apoptosis or necrosis is achieved.

Original languageEnglish (US)
Pages (from-to)9-20
Number of pages12
JournalCirculation Research
Volume84
Issue number1
Publication statusPublished - Jan 8 1999
Externally publishedYes

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Keywords

  • Myocardial ischemia
  • Myofilament
  • Protein degradation
  • Transglutaminase
  • Troponin I

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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