A 16 kDa protein substrate for protein kinase C and its phosphorylation upon stimulation of vasopressin receptors in rat aortic myocytes

Abstract

Phosphorylation induced by protein kinase C was examined in a plasma membrane fraction from rat aortic myocytes. Labelled phosphate incorporation produced by addition of kinase C to the membrane preparation allowed to identify a 16 kDa protein as the major substrate of the enzyme. This protein electrophoretically migrated with a protein phosphorylated by cAMP dependent protein kinase, but the two kinases produced phosphorylation of different sites since their effects were additive. Pretreatment of the myocytes with a kinase C activating phorbol ester or with vasopressin decreased further phosphate incorporation into the 16 kDa protein under the influence of exogenous kinase C. The results provide evidence that vasopressin produced in situ phosphorylation of the 16 kDa protein in rat aortic myocytes, with a time course and at concentrations consistent with a role of kinase C activation in the response of aortic myocytes to stimulation of V 1 receptors.