title: Interaction of calmodulin with myosin light chain kinase and cAMP-dependent protein kinase in bovine brain.
abstract: Myosin light chain kinase and a fraction of type II cAMP-dependent protein kinase have been partially purified from bovine brain by affinity chromatography on calmodulin-Sepharose. The myosin kinase was purified approximately 3700-fold and has an estimated molecular weight of 130,000 +/- 10,000 by sodium dodecyl sulfate gel electrophoresis. A fraction of soluble cAMP-dependent protein kinase also bound to calmodulin-Sepharose and was purified 2300-fold. A fraction of this cAMP-dependent protein kinase after purification by glycerol gradient centrifugation was shown to contain the two subunits of calcineurin, a major calmodulin-binding protein in brain, and the two subunits of type II cAMP-dependent protein kinase in a ratio of 1:1:2:2. Its sedimentation coefficient was 8.1 S and 9.0 S when centrifuged in the absence or presence of calmodulin, suggesting the formation of a complex between calmodulin and protein kinase. Our results suggest the possibility that calcineurin may be involved in the interaction between the protein kinase and calmodulin. Furthermore, our studies imply that the regulatory subunit of the cAMP-dependent protein kinase, but not the catalytic subunit, is the site of interaction with calmodulin since the catalytic subunit of protein kinase was partially resolved from the complex by cAMP.
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002417:Cattle; D002846:Chromatography, Affinity; D000242:Cyclic AMP / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009219:Myosin-Light-Chain Kinase; D011485:Protein Binding; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D002147:Calmodulin; D046911:Macromolecular Substances; D000242:Cyclic AMP; D011494:Protein Kinases; D009219:Myosin-Light-Chain Kinase