Stage-specific changes in myofilament protein phosphorylation following myocardial infarction in mice.

The response of cardiac muscle to an insult such as myocardial infarction includes changes in the expression of numerous signaling proteins and modulation of gene expression, as well as post-translational modifications of existing proteins. Most studies to date have defined these in end-stage cardiac muscle thus obviating consideration of the temporal progression that causes the heart to transition from a compensated to a decompensated phenotype. To explore these transitions, we examined contractile protein biochemistry in a mouse MI model at two early time points: 2 days and 2 weeks post-infarct and at two later time points: 2 and 4 months post-infarct. Phosphorylation of myofilament proteins was analyzed using phosphospecific staining of polyacrylamide gels, and whenever possible, phosphospecific antibodies. Phosphorylation of myosin binding protein c, the myosin regulatory light chain and troponin I were all decreased relative to sham operated animals at both early time points. However, by 2 months, total phosphorylation of all the major myofilament proteins normalized and at both 2 and 4 months, there was a significant increase in troponin I phosphorylation. One-dimensional IEF of troponin I coupled with phospho-specific antibody analysis demonstrated a redistribution of phosphorylation sites with a significant initial decline at the putative PKA sites, Serine 22,23, and a subsequent increase at the putative PKC site, serine 43,45. These data suggest that temporal changes in myofilament protein phosphorylation contribute both to the initial compensatory hyperdynamic response to myocardial infarction and subsequently to the gradual progression to myocardial failure. (c) 2009 Elsevier Ltd. All rights reserved.