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(Circulation Research. 2001;88:291.)
© 2001 American Heart Association, Inc.

Cellular Biology

AKAP-Mediated Targeting of Protein Kinase A Regulates Contractility in Cardiac Myocytes

Mary A. Fink, Daniel R. Zakhary, Julie A. Mackey, Russell W. Desnoyer, Carolyn Apperson-Hansen, Derek S. Damron, Meredith Bond

From the Department of Molecular Cardiology (M.A.F., D.R.Z., R.W.D., M.B.), Department of Biostatistics and Epidemiology (C.A.-H.), and Center for Anesthesiology Research (D.S.D.), Cleveland Clinic Foundation, and Department of Physiology and Biophysics (D.R.Z., J.A.M., M.B.), School of Medicine, Case Western Reserve University, Cleveland, Ohio.

Correspondence to Meredith Bond, Department of Molecular Cardiology, The Cleveland Clinic Foundation, 9500 Euclid Ave, Cleveland, OH 44195. E-mail bondm{at}ccf.org

Abstract—Compartmentalization of cAMP-dependent protein kinase A (PKA) by A-kinase anchoring proteins (AKAPs) targets PKA to distinct subcellular locations in many cell types. However, the question of whether AKAP-mediated PKA anchoring in the heart regulates cardiac contractile function has not been addressed. We disrupted AKAP-mediated PKA anchoring in cardiac myocytes by introducing, via adenovirus-mediated gene transfer, Ht31, a peptide that binds the PKA regulatory subunit type II (RII) with high affinity. This peptide competes with endogenous AKAPs for RII binding. Ht31P (a proline-substituted derivative), which does not bind RII, was used as a negative control. We then investigated the effects of Ht31 expression on RII distribution, Ca²⁺ cycling, cell shortening, and PKA-dependent substrate phosphorylation. By confocal microscopy, we showed redistribution of RII from the perinuclear region and from periodic transverse striations in Ht31P-expressing cells to a diffuse cytosolic localization in Ht31-expressing cells. In the presence of 10 nmol/L isoproterenol, Ht31-expressing myocytes displayed an increased rate and amplitude of cell shortening and relaxation compared with control cells (uninfected and Ht31P-expressing myocytes); with isoproterenol stimulation we observed decreased time to 90% decline in Ca²⁺ but no significant difference between Ht31-expressing and control cells in the rate of Ca²⁺ cycling or amplitude of the Ca²⁺ transient. The increase in PKA-dependent phosphorylation of troponin I and myosin binding protein C on isoproterenol stimulation was significantly reduced in Ht31-expressing cells compared with controls. Our results demonstrate that, in response to ß-adrenergic stimulation, cardiomyocyte function and substrate phosphorylation by PKA is regulated by targeting of PKA by AKAPs.

Key Words: A-kinase anchoring proteins • protein kinase A • cardiac myocyte • ß-adrenergic receptor • contractility

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