Cyclic AMP-Dependent Activation of Protein Kinases in the Myocardium

  • J. G. DobsonJr.


Upon catecholamine stimulation of the heart, the cyclic AMP-dependent activation of protein kinases appears to play an important role in the enhancement of contractility and glycogenolysis presumably by promoting the phosphorylation of myocardial proteins. In the intact rat heart, epinephrine produced a rapid elevation of myocardial cyclic AMP and activation of both cyclic AMP-dependent protein kinase and phosphorylase b kinase within 5–10 sec. This preceded the conversion of phosphorylase b to phosphorylase a and the augmentation of contractile state that occurred within 12–18 sec. The β-adrenergic blocking agent, practolol, had no effect on the basal levels of the above parameters, but did prevent the epinephrine produced increases. The importance of catecholamine activation of the kinases in catalyzing protein phosphorylation was investigated in isolated myocardial preparations previously exposed to 32P to label intracellular ATP. In ventricular muscle slices, isoproterenol increased the 32P incorporation into trichloroacetic acid insoluble protein as well as the phosphate content of this protein. In isolated perfused hearts, the catecholamine produced an increase in 32P incorporation into a 29000Xg (membrane and mitochondria) and 100 000 × g (sarcoplasmic reticulum) pellet fractions and a 100 000 × g soluble fraction within 30 sec. While the β-adrenergic blocking agent, propranolol, did not by itself affect 32P incorporation, it prevented the isoproterenol-induced incorporation of 32P into the pellet and soluble fractions and the acid-insoluble protein. Removal of isoproterenol from the isolated preparations eliminated the differences in 32P incorporation indicating that the effects of the catecholamines were reversible. Sodium dodecyl sulfate gel electrophoresis of ventricular muscle homogenates of isolated perfused hearts revealed that isoproterenol enhanced the 32P incorporation into several myocardial proteins having molecular weights of 155 000, 92 000, 30 000, 16 000 and 13 000 within 30 sec. These findings are consistent with the notion that cyclic AMP-dependent activation of protein kinases by catecholamine stimulation enhances the phosphorylation of myocardial proteins which in turn may play an important role in the augmentation of contractility and glycogenolysis.


Perfuse Heart Contractile State Cardiac Sarcoplasmic Reticulum Glycogen Phosphorylase Activity Myocardial Protein 
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© Springer-Verlag, Berlin, Heidelberg 1981

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  • J. G. DobsonJr.

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