Regulation by phosphorylase kinase of phosphoprotein phosphatase activity: simultaneous control of protein phosphorylation and dephosphorylation in skeletal muscle.

Phosphorylase kinase from rabbit skeletal muscle inhibited the dephosphorylation of phosphorylase a by phosphoprotein phosphatase. Phosphorylation (activation) of phosphorylase kinase by cyclic AMP-dependent protein kinase greatly increased this inhibitory effect. Thus, phosphoprotein phosphatase is inhibited by phosphorylase kinase in a reversible manner (Gergely et al. (1976) Biochim. Biophys. Acta 429 809-816). In this paper the regulation by phosphorylase kinase at phosphoprotein phosphatase activity in different fractions of muscle extract and in the presence of various ligands has been investigated. The presence of phosphorylase kinase also affected the ligand control of phosphatase activity. Phosphorylase kinase almost cancelled the inhibitory effect of AMP but hardly influenced the activating effect of glucose, glucose 6-phosphate and caffeine. Calmodulin, glycogen and phosphorylase b (effectors of phosphorylase kinase) did not influence the inhibitory effect of phosphorylase kinase. Fractions of muscle extract also demonstrated the regulatory role of phosphorylase kinase. These fractions contained considerable amounts of phosphorylase kinase and phosphatase. Phosphatase activity was inhibited by phosphorylation reactions triggered by Mg++ and ATP. Heat-stable inhibitors were absent from these fractions, therefore the transient inhibition of phosphatase could be attributed to the phosphorylation of endogenous phosphorylase kinase. The introduction between phosphorylase kinase and phosphatase resulted in a loss of AMP sensitivity, i.e. AMP did not inhibit the activity of phosphatase in those fractions. Our results imply that the phosphorylation of phosphorylase kinase is equally important both in the formation of enzymatically active phosphorylase a and in the inhibition of dephosphorylation of phosphorylase a. The consequence of these two effects is the elevated level of phosphorylase a.