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Myocardial infarction in mice alters sarcomeric function via post-translational protein modification

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Abstract

Myocardial physiology in the aftermath of myocardial infarction (MI) before remodeling is an under-explored area of investigation. Here, we describe the effects of MI on the cardiac sarcomere with focus on the possible contributions of reactive oxygen species. We surgically induced MI in 6–7-month-old female CD1 mice by ligation of the left anterior descending coronary artery. Data were collected 3–4 days after MI or sham (SH) surgery. MI hearts demonstrated ventricular dilatation and systolic dysfunction upon echo cardiographic analysis. Sub-maximum Ca-activated tension in detergent-extracted fiber bundles from papillary muscles increased significantly in the preparations from MI hearts. Ca2+ sensitivity increased after MI, whereas cooperativity of activation decreased. To assess myosin enzymatic integrity we measured splitting of Ca-ATP in myofibrillar preparations, which demonstrated a decline in Ca-ATPase activity of myofilament myosin. Biochemical analysis demonstrated post-translational modification of sarcomeric proteins. Phosphorylation of cardiac troponin I and myosin light chain 2 was reduced after MI in papillary samples, as measured using a phospho-specific stain. Tropomyosin was oxidized after MI, forming disulfide products detectable by diagonal non-reducing–reducing SDS-PAGE. Our analysis of myocardial protein oxidation post-MI also demonstrated increased S-glutathionylation. We functionally linked protein oxidation with sarcomere function by treating skinned fibers with the sulfhydryl reducing agent dithiothreitol, which reduced Ca2+ sensitivity in MI, but not SH, samples. Our data indicate important structural and functional alterations to the cardiac sarcomere after MI, and the contribution of protein oxidation to this process.

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Acknowledgments

We thank Dr. Marcelo Bonini for his helpful advice on assessing glutathionylation. This study was supported by American Heart Association-Midwest Pre-Doctoral Fellowship 2008-06682-00-00 (BSA) and by NIH Grants T32 007692 (BSA, KMS, SBS) and PO1 HL062426 (RJS, DLG, PHG).

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Authors and Affiliations

  1. Department of Physiology and Biophysics, (M/C 901), College of Medicine, University of Illinois at Chicago, 835 S. Wolcott Ave., Chicago, IL, 60612–7342, USA

    Benjamin S. Avner, Sarah B. Scruggs, Mariam Farjah & R. John Solaro

  2. Section of Cardiology, Department of Medicine, University of Illinois at Chicago, Chicago, IL, USA

    Krystyna M. Shioura, Milana Grachoff, David L. Geenen & Paul H. Goldspink

  3. Chicago Biomedical Consortium/Research Resources Center Proteomics and Informatics Services Facility, University of Illinois at Chicago, Chicago, IL, USA

    Donald L. Helseth Jr.

  4. Section of Cardiology, Departments of Physiology and Medicine, University of California at Los Angeles, Los Angeles, CA, USA

    Sarah B. Scruggs

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  1. Benjamin S. Avner
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Correspondence to R. John Solaro.

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Avner, B.S., Shioura, K.M., Scruggs, S.B. et al. Myocardial infarction in mice alters sarcomeric function via post-translational protein modification. Mol Cell Biochem 363, 203–215 (2012). https://doi.org/10.1007/s11010-011-1172-z

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  • DOI: https://doi.org/10.1007/s11010-011-1172-z

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