Dephosphorylation and inactivation of phosphorylase kinase: subunit specificity of rabbit skeletal muscle protein phosphatases.
The dephosphorylation of phosphorylase kinase by four rabbit skeletal muscle protein phosphatases was studied. The four enzymes used were preparations of protein phosphatases C-I, C-II, H-I, and H-II. Phosphatases C-I, C-II, and H-II were obtained as homogeneous preparations using procedures previously developed. Phosphatase H-I was purified 644-fold from rabbit skeletal muscle for the purposes of this study, and was the major phosphorylase phosphatase activity in the tissue extract. Phosphatases C-I and H-I were relatively specific for removal of the beta subunit phosphate of phosphorylase kinase, this occurring at rates approximately 100 times more rapidly than the removal of the alpha subunit phosphate. In contrast, phosphatases C-II and H-II readily dephosphorylated both the alpha and beta subunits, although the alpha subunit phosphate release occurred at rates about twice that of the beta subunit phosphate. These studies show that skeletal muscle contains two phosphatases capable of acting on phosphorylase kinase, and that these have different specificities as represented by phosphatases H-I and C-I on the one hand, and phosphatases C-II and H-II on the other hand. These studies also provided unequivocal evidence that dephosphorylation of the beta subunit of phosphorylase kinase is solely involved in the inactivation of the cAMP-dependent protein kinase-activated enzyme. When autophosphorylated phosphorylase kinase was used as the substrate, the four phosphatases displayed similar general specificities as they did toward the cAMP-dependent protein kinase-activated enzyme. With none of the phosphatases examined was there any evidence that alpha subunit phosphorylation affected the rate of beta subunit dephosphorylation.
D000818:Animals; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism*; D010764:Phosphorylase Kinase / Q000037:antagonists & inhibitors*; D010765:Phosphorylase Phosphatase / Q000302:isolation & purification / Q000378:metabolism; D010762:Phosphorylase a / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity
D046911:Macromolecular Substances; D010762:Phosphorylase a; D010764:Phosphorylase Kinase; C022410:phosphorylase kinase phosphatase; D010749:Phosphoprotein Phosphatases; D010765:Phosphorylase Phosphatase