Regulation of calcium transport in cardiac sarcoplasmic reticulum by cyclic AMP-dependent protein kinase.
A manyfold increase in phosphorylation of cardiac sarcoplasmic reticulum (SR) was seen when SR was incubated in the presence of a bovine cardiac cyclic AMP-dependent protein kinase and cyclic AMP. This phosphoprotein had stability characteristics of a phosphoester in which the phosphate is incorporated largely into serine, and its formation did not required calcium ions, unlike the formation of acyl phosphoprotein intermediate of calcium-transport ATPase which is present within the same membrane. When examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein kinase-catalyzed phosphorylation occurred at a 22,000-dalton component of the cardiac sarcoplasmic reticulum. This 22,000-dalton protein has been named "phospholamban" (lambda alpha mu beta alpha nu epsilon iota nu = to receive), based on its ability to receive phosphate from ATP. Phosphorylation of phospholamban by cyclic AMP-dependent protein kinase was associated with the stimulation of calcium transport by the cardiac sarcoplasmic reticulum. This stimulation was accompanied by an increase in the calcium-activated ATPase activity, indicating that the overall rate of calcium transport rather than its efficiency is enhanced by protein kinase. The 22,000-dalton phopholamban was susceptible to trypsin. Brief digestion with trypsin in the presence of 1 M sucrose prevented subsequent phosphorylation of phospholamban, while leaving the calcium pump apparently intact. Incubation of trypsin-treated sarcoplasmic reticulum with cyclic AMP-depentent protein kinase did not result in the stimulation of calcium transport. These results may suggest that phospholamban is a modulator of the calcium pump of the cardiac sarcoplasmic reticulum.
D000818:Animals; D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology*; D004285:Dogs; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008954:Models, Biological; D008970:Molecular Weight; D009124:Muscle Proteins / Q000378:metabolism; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*
D009124:Muscle Proteins; D010750:Phosphoproteins; D000242:Cyclic AMP; D011494:Protein Kinases; D002118:Calcium