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Open Access 01-03-2010 | Original Contribution

Protein kinase C α and ε phosphorylation of troponin and myosin binding protein C reduce Ca²⁺ sensitivity in human myocardium

Authors: Viola Kooij, Nicky Boontje, Ruud Zaremba, Kornelia Jaquet, Cris dos Remedios, Ger J. M. Stienen, Jolanda van der Velden

Published in: Basic Research in Cardiology | Issue 2/2010

Abstract

Previous studies indicated that the increase in protein kinase C (PKC)-mediated myofilament protein phosphorylation observed in failing myocardium might be detrimental for contractile function. This study was designed to reveal and compare the effects of PKCα- and PKCε-mediated phosphorylation on myofilament function in human myocardium. Isometric force was measured at different [Ca²⁺] in single permeabilized cardiomyocytes from failing human left ventricular tissue. Activated PKCα and PKCε equally reduced Ca²⁺ sensitivity in failing cardiomyocytes (ΔpCa₅₀ = 0.08 ± 0.01). Both PKC isoforms increased phosphorylation of troponin I- (cTnI) and myosin binding protein C (cMyBP-C) in failing cardiomyocytes. Subsequent incubation of failing cardiomyocytes with the catalytic subunit of protein kinase A (PKA) resulted in a further reduction in Ca²⁺ sensitivity, indicating that the effects of both PKC isoforms were not caused by cross-phosphorylation of PKA sites. Both isozymes showed no effects on maximal force and only PKCα resulted in a modest significant reduction in passive force. Effects of PKCα were only minor in donor cardiomyocytes, presumably because of already saturated cTnI and cMyBP-C phosphorylation levels. Donor tissue could therefore be used as a tool to reveal the functional effects of troponin T (cTnT) phosphorylation by PKCα. Massive dephosphorylation of cTnT with alkaline phosphatase increased Ca²⁺ sensitivity. Subsequently, PKCα treatment of donor cardiomyocytes reduced Ca²⁺ sensitivity (ΔpCa₅₀ = 0.08 ± 0.02) and solely increased phosphorylation of cTnT, but did not affect maximal and passive force. PKCα- and PKCε-mediated phosphorylation of cMyBP-C and cTnI as well as cTnT decrease myofilament Ca²⁺ sensitivity and may thereby reduce contractility and enhance relaxation of human myocardium.

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Title: Protein kinase C α and ε phosphorylation of troponin and myosin binding protein C reduce Ca2+ sensitivity in human myocardium
Authors: Viola Kooij
Nicky Boontje
Ruud Zaremba
Kornelia Jaquet
Cris dos Remedios
Ger J. M. Stienen
Jolanda van der Velden
Publication date: 01-03-2010
Publisher: D. Steinkopff-Verlag
Published in: Basic Research in Cardiology / Issue 2/2010
Print ISSN: 0300-8428
Electronic ISSN: 1435-1803
DOI: https://doi.org/10.1007/s00395-009-0053-z

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Protein kinase C α and ε phosphorylation of troponin and myosin binding protein C reduce Ca²⁺ sensitivity in human myocardium

Abstract

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