The ATPase activity of phosphorylase kinase is regulated in parallel with its protein kinase activity.
Phosphorylase kinase from rabbit skeletal muscle has been found to have an intrinsic ATPase activity that occurs at a rate approximately 0.2% of that of its phosphorylase conversion activity and about three times that of its autophosphorylation activity. The characteristics of this ATPase activity were in all aspects tested essentially the same as the kinase's phosphorylase conversion activity. The ATPase requires Mg2+ and is dramatically stimulated by Ca2+ ions. At neutral pH there is a pronounced lag in the rate of product formation that is not present at alkaline pH, a condition that greatly stimulates both the phosphorylase conversion and ATPase activities. ATP is preferentially hydrolyzed over GTP and the Km for MgATP determined in the ATPase assay is 0.14 mM. ADP, an allosteric activator of phosphorylase conversion, also stimulates the ATPase activity, whereas beta-glycerophosphate, an inhibitor of phosphorylase conversion, is an inhibitor of the ATPase activity. Phosphorylation or partial proteolysis of the kinase, which are known to activate phosphorylase conversion, also activate the ATPase activity. Because the phosphorylase conversion and ATPase activities are regulated in parallel, we conclude that activation of the two catalytic activities must share a common underlying basis, namely an enhanced phosphotransferase activity that is independent of the phosphoryl acceptor.
Paudel
H K
HK
Department of Biochemistry, College of Medicine, University of Tennessee, Memphis 38163.
Carlson
G M
GM
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D004533:Egtazic Acid; D004789:Enzyme Activation; D005994:Glycerophosphates / Q000378:metabolism; D007700:Kinetics; D010764:Phosphorylase Kinase / Q000378:metabolism*; D010766:Phosphorylation; D011487:Protein Conformation; D011494:Protein Kinases / Q000378:metabolism*; D011817:Rabbits
D005994:Glycerophosphates; D004533:Egtazic Acid; D000244:Adenosine Diphosphate; D011494:Protein Kinases; D010764:Phosphorylase Kinase; D000251:Adenosine Triphosphatases; D002118:Calcium; C031463:beta-glycerophosphoric acid