Studies on phosphorylation of canine cardiac sarcoplasmic reticulum by calmodulin-dependent protein kinase.
Two endogenous protein kinase activities, cAMP-dependent and calmodulin-Ca2+-dependent, are associated with isolated cardiac sarcoplasmic reticulum (SR) vesicles. Both kinases phosphorylate an endogenous substrate of approximately 22,000 daltons (phospholamban). The phosphorylation of phospholamban by either the intrinsic or by exogenous cAMP-dependent protein kinase is found to be Ca2+-independent between 0.05 and 100 microM free Ca2+. Calmodulin-dependent phosphorylation, on the other hand, does not require cAMP and is absolutely dependent on the presence of free Ca2+ over a concentration range that corresponds to physiological levels (10(-7) to 10(-5) M). Phosphorylation of SR vesicles by both kinases is additive and the extent of saturation of the cAMP-specific sites has no effect on the degree of stimulation by calmodulin or its Ca2+-dependence. Trifluoperazine, an inhibitor of calmodulin, inhibits calmodulin-dependent phosphorylation without affecting cAMP-dependent phosphorylation, indicating the presence of two types of kinases. This is made further evident by the selectivity of each kinase for exogenous substrates. Whereas cAMP-dependent protein kinase appears to phosphorylate histone ILA (a basic protein) preferentially, calmodulin-dependent protein kinase prefers phosvitin (an acidic protein).
D000251:Adenosine Triphosphatases / Q000494:pharmacology; D000818:Animals; D001345:Autoradiography; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D002417:Cattle; D000242:Cyclic AMP / Q000378:metabolism; D004285:Dogs; D004533:Egtazic Acid / Q000494:pharmacology; D009206:Myocardium / Q000201:enzymology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D014268:Trifluoperazine / Q000494:pharmacology; D014336:Troponin / Q000494:pharmacology; D000099211:Phospholamban
D000251:Adenosine Triphosphatases; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D000242:Cyclic AMP; D004533:Egtazic Acid; D011494:Protein Kinases; D014268:Trifluoperazine; D014336:Troponin; D000099211:Phospholamban