A calmodulin dependent protein kinase activity associated with rabbit heart sarcolemma.
Sarcolemmal vesicles prepared from rabbit heart muscle by differential and discontinuous sucrose density gradient centrifugation, exhibited high marker enzyme activities: Na+ + K+ ATPase 22 microMol Pi x mg-1 x h-1, Adenylate cyclase 500 pmole cAMP x mg-1 x min-1, calcium antagonist receptors 0.7 pmoles x mg-1. Calmodulin in the presence of calcium and gamma-ATP32 stimulated rapidly and specifically the 32P incorporation into two membrane proteins of 54 and 44 kDa. Calmodulin stimulated the phosphorylation of the 44 kDa to a greater extent (17.9 pmol 32P x mg-1 protein) than the 54 kDa protein (1.3 pmoles 32P x mg-1 protein). Removal of endogenous calmodulin from the membrane by EGTA extraction resulted in a 2.5 fold increase in calmodulin dependent 32P incorporation into the two proteins in the presence of exogenous calmodulin. It is suggested that the calmodulin dependent protein kinase activity in heart sarcolemma may mediate the effects of calmodulin in the regulation of Ca2+ transport across the membrane.
Tuana
B S
BS
Department of Pharmacology, University of Toronto, Ontario, Canada.
Murphy
B J
BJ
Schwarzkopf
C
C
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002147:Calmodulin / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D006321:Heart / Q000187:drug effects; D066298:In Vitro Techniques; D007700:Kinetics; D008970:Molecular Weight; D009124:Muscle Proteins / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology* / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011817:Rabbits; D012508:Sarcolemma / Q000187:drug effects / Q000201:enzymology*; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology
D002147:Calmodulin; D009124:Muscle Proteins; D000255:Adenosine Triphosphate; D011494:Protein Kinases