The effect of PKA-mediated phosphorylation of ryanodine receptor on SR Ca²⁺ leak in ventricular myocytes.

ABSTRACT: Functional impact of cardiac ryanodine receptor (type 2 RyR or RyR2) phosphorylation by protein kinase A (PKA) remains highly controversial. In this study, we characterized a functional link between PKA-mediated RyR2 phosphorylation level and sarcoplasmic reticulum (SR) Ca²⁺ release and leak in permeabilized rabbit ventricular myocytes. Changes in cytosolic [Ca²⁺] and intra-SR [Ca²⁺]_SR were measured with Fluo-4 and Fluo-5N, respectively. Changes in RyR2 phosphorylation at two PKA sites, serine-2031 and -2809, were measured with phospho-specific antibodies. cAMP (10μM) increased Ca²⁺ spark frequency approximately two-fold. This effect was associated with an increase in SR Ca²⁺ load from 0.84 to 1.24mM. PKA inhibitory peptide

SUBMITTER: Bovo E 

PROVIDER: S-EPMC5337148 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature