Calcium has a direct effect on thick filament activation in porcine myocardium

Mohran, Saffie; McMillen, Timothy S.; Mandrycky, Christian; Tu, An-Yue; Kooiker, Kristina B.; Qian, Wenjing; Neys, Stephanie; Osegueda, Brayan; Moussavi-Harami, Farid; Irving, Thomas C.; Regnier, Michael; Ma, Weikang

doi:10.1085/jgp.202413545

Title: Calcium has a direct effect on thick filament activation in porcine myocardium

Journal Article · 20 September 2024 · Journal of General Physiology

DOI: https://doi.org/10.1085/jgp.202413545 · OSTI ID:2447492

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Sarcomere activation in striated muscle requires both thin filament–based and thick filament–based activation mechanisms. Recent studies have shown that myosin heads on the thick filaments undergo OFF to ON structural transitions in response to calcium (Ca2+) in permeabilized porcine myocardium in the presence of a small molecule inhibitor that eliminated active force. The changes in X-ray diffraction signatures of OFF to ON transitions were interpreted as Ca2+ acting to activate the thick filaments. Alternatively, Ca2+ binding to troponin could initiate a Ca2+-dependent crosstalk from the thin filament to the thick filament via interfilament connections such as the myosin binding protein-C. Here, we exchanged native troponin in permeabilized porcine myocardium for troponin containing the cTnC D65A mutation, which disallows the activation of troponin through Ca2+ binding to determine if Ca2+-dependent thick filament activation persists in the absence of thin filament activation. After the exchange protocol, over 95% of the Ca2+-activated force was eliminated. Equatorial intensity ratio increased significantly in both WT and D65A exchanged myocardium with increasing Ca2+ concentration. The degree of helical ordering of the myosin heads decreased by the same amount in WT and D65A myocardium when Ca2+ concentration increased. These results are consistent with a direct effect of Ca2+ in activating the thick filament rather than an indirect effect due to Ca2+-mediated crosstalk between the thick and thin filaments.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 2447492

Journal Information:: Journal of General Physiology, Journal Name: Journal of General Physiology Journal Issue: 11 Vol. 156; ISSN 0022-1295

Publisher:: Rockefeller University PressCopyright Statement

Country of Publication:: United States

Language:: English

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