A-kinase Anchoring Protein 100 (AKAP100) is Localized in Multiple Subcellular Compartments in the Adult Rat Heart

doi:10.1083/jcb.142.2.511

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J. Cell Biol., Volume 142, Number 2, July 27, 1998 511-522

A-kinase Anchoring Protein 100 (AKAP100) is Localized in Multiple Subcellular Compartments in the Adult Rat Heart

Jiacheng Yang,^* Judith A. Drazba, Donald G. Ferguson,^§ and Meredith Bond^*

^* Department of Molecular Cardiology and Department of Neuroscience, The Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195; and ^§ Department of Anatomy and Department of Physiology and Biophysics, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106

Stimulation of $beta$ -adrenergic receptors activates type I and II cyclic AMP-dependent protein kinase A, resulting in phosphorylationof various proteins in the heart. It has been proposed that PKAII compartmentalization by A-kinase-anchoring proteins (AKAPs)regulates cyclic AMP-dependent signaling in the cell. We investigatedthe expression and localization of AKAP100 in adult hearts. Byimmunoblotting, we identified AKAP100 in adult rat and human hearts,and showed that type I and II regulatory (RI and II) subunitsof PKA are present in the rat heart. By immunofluorescence andconfocal microscopy of rat cardiac myocytes and cryostat sectionsof rat left ventricle papillary muscles, we localized AKAP100to the nucleus, sarcolemma, intercalated disc, and at the levelof the Z-line. After double immunostaining of transverse cross-sectionsof the papillary muscles with AKAP100 plus $alpha$ -actinin-specificantibodies or AKAP100 plus ryanodine receptor-specific antibodies,confocal images showed AKAP100 localization at the region of thetransverse tubule/junctional sarcoplasmic reticulum. RI is distributeddifferently from RII in the myocytes. RII, but not RI, was colocalizedwith AKAP100 in the rat heart. Our studies suggest that AKAP100tethers PKA II to multiple subcellular compartments for phosphorylationof different pools of substrate proteins in the heart.

Key words: AKAP; PKA; RI; RII; heart

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