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Calcium regulation in the human myocardium affected by dilated cardiomyopathy: A structural basis for impaired Ca2+-sensitivity

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Abstract

Calcium regulation in the human heart is impaired during idiopathic dilated cardiomyopathy (IDC). Here, we analyze the structural basis for impairment in the regulatory mechanism. Regulation of contractility was monitored by MgATPase and Ca2+-binding assays as a function of calcium. Myofibrillar proteolysis and expression of troponin T isoforms were established by gel electrophoresis and by Western blots. Myofibrillar ATPase assays in low salt however, revealed a drastic lowering of calcium sensitivity in IDC myofibrils as indicated by reductions in both activation by high calcium and in EGTA-mediated inhibition of MgATPase. Structural changes in myofilament proteins were found in most IDC hearts, specifically proteolysis of myosin light chain 2 (LC2), troponin T and I (TnT and TnI), and sometimes large isoform shift in TnT. IDC did not induce mutations in LC2 and troponin C (TnC), as established by cDNA sequence data from IDC cases, thus, calcium binding to IDC myofibrils was unaffected. Reassociation of IDC myofibrils with native LC2 raised MgATPase activation at high Ca2+ to control levels, while repletion with intact, canine TnI/TnT restored inhibition at low Ca2+. A model, identifying possible steps in the steric blocking mechanism of regulation, is proposed to explain IDC-induced changes in Ca2+-regulation. Moreover, shifts in TnT isoforms may imply either a genetic or a compensatory factor in the development and pathogenesis of some forms of IDC.

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Authors and Affiliations

  1. Department of Medicine and Biochemistry and Molecular Biology, Albany Medical College, Albany, USA E-mail: SMargossian@worldnet.att.net

    Sarkis S. Margossian & Mark Deziel

  2. Division of Cardiology, Duke University Medical Center, Durham, USA

    Page A.W. Anderson

  3. Department of Basic Veterinary Sciences, Royal Veterinary College, London, UK

    Peter D. Chantler

  4. Department of Pathology, University of Alabama in Birmingham, Birmingham, USA

    Patrick K. Umeda & Ashwani Malhotra

  5. Boston Biomedical Research Institute, Boston, USA

    Paul Norton

  6. Department of Medicine, Albert Einstein College of Medicine, Bronx, USA

    Fan Yang

  7. Department of Pathology, Albany Medical College, Albany, USA

    James B. Caulfield & Henry S. Slayter

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  1. Sarkis S. Margossian
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  2. Page A.W. Anderson
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Margossian, S.S., Anderson, P.A., Chantler, P.D. et al. Calcium regulation in the human myocardium affected by dilated cardiomyopathy: A structural basis for impaired Ca2+-sensitivity. Mol Cell Biochem 194, 301–313 (1999). https://doi.org/10.1023/A:1006980405359

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