Peroxynitrite-induced inhibition and nitration of cardiac myofibrillar creatine kinase

Michael J. Mihm, John Anthony Bauer

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

Although cardiac peroxynitrite formation and attendant protein nitration is an established event in both acute and chronic settings of cardiac failure, the putative intracellular targets involved remain incompletely defined. We have recently shown that the myofibrillar isoform of creatine kinase (a critical energetic controller of cardiomyocyte contractility) may be a particularly sensitive target of peroxynitrite-induced nitration and inactivation in vivo. However, the kinetic and mechanistic aspects of this interaction remain undefined. Here we tested the hypothesis that myofibrillar creatine kinase is sensitive to inhibition by peroxynitrite, and investigated the mechanistic role for tyrosine nitration in this process. Peroxynitrite potently and irreversibly inhibited myofibrillar creatine kinase capacity (Vmax), at concentrations as low as 100 nM, while substrate affinity (Km) was unaffected. Concentration-dependent nitration of myofibrillar creatine kinase was observed. The extent of nitration was linearly related to peroxynitrite concentration and highly correlated to the extent of myofibrillar creatine kinase inhibition. This inhibition was not reversible by treatment with free cysteine (250 μM), but pre-incubation with substrate (phosphocreatine and/or ATP) provided significant protection of MM-CK from both nitration and inhibition. These results suggest that myofibrillar creatine kinase is a highly sensitive target of peroxynitrite-mediated inhibition, and that nitration may mediate this inhibition.

Original languageEnglish
Pages (from-to)1013-1019
Number of pages7
JournalBiochimie
Volume84
Issue number10
DOIs
StatePublished - Oct 1 2002

Bibliographical note

Funding Information:
This work was supported in part by grants from the National Institutes of Health (HL59791, DK55053, HL63067) and an award from the American Heart Association, Ohio Valley Affiliate.

Keywords

  • 3-nitro-1-tyrosine
  • Heart failure
  • Myofibrillar creatine kinase
  • Peroxynitrite
  • Protein nitration

ASJC Scopus subject areas

  • Biochemistry

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