The effect of pH on the Ca2+ affinity of the Ca2+ regulatory sites of skeletal and cardiac troponin C in skinned muscle fibres

Bruce Parsons, Danuta Szczesna, Jiaju Zhao, Glen Van Slooten, W. Glenn L. Kerrick, John A. Putkey, James D. Potter

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

It is known that intracellular pH drops rapidly after the onset of ischemia in cardiac muscle and may play some role in the rapid drop in force that ensues. It is also known that α1-adrenoceptor agonists alkalinize intracellular pH by stimulating Na+/H+ exchange and may represent a mechanism which facilitates recovery of intracellular pH from acidosis. Lowering or raising pH shifts the Ca2+ dependence of force development in muscle fibres to higher or lower free Ca2+ concentrations, respectively, yet the precise mechanism is unknown. To investigate this phenomenon we have used skinned skeletal or cardiac muscle fibres whose endogenous troponin C (TnC) has been replaced with chicken skeletal TnC labelled with DANZ (STnC(DANZ)) or recombinant cardiac TnC labelled with IAANS (CTnC3(C84)(IAANS)), respectively. The fluorescence of the STnC(DANZ) or CTnC3(C84)(IAANS) was enhanced by Ca2+ binding to the Ca2+-specific (regulatory) site(s) of STnC or CTnC when incorporated into skinned fibres, and was measured simultaneously with force. When the pH was changed from 7.0 to 6.5 or 7.5 the shift in the Ca2+ dependence of force paralleled the shift in fluorescence. Since the force and fluorescence shift in parallel as the pH is lowered or raised, it can be concluded that these changes in Ca2+ sensitivity are caused by a decrease or increase, respectively, in the Ca2+ affinity of the Ca2+-specific site(s) of TnC. Since lowering or raising the pH also resulted in lower or higher, respectively, maximal Ca2+ activated force while maximal fluorescence remained unchanged, it is possible that H+ may act indirectly, as well, by reducing or increasing, respectively, the number or type of crossbridges attached to actin and thereby alter the crossbridge induced depression or elevation, respectively of the observed TnC Ca2+ affinity. Experiments with 2,3-butanedione monoxime, however, where force-generating crossbridges were greatly reduced, indicated that the pH effect may be primarily related to a direct change in the Ca2+ affinity to the regulatory sites of TnC.

Original languageEnglish (US)
Pages (from-to)599-609
Number of pages11
JournalJournal of Muscle Research and Cell Motility
Volume18
Issue number5
DOIs
StatePublished - Oct 18 1997

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Cell Biology

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