title: Effects of myosin light chain kinase and peptides on Ca2+ exchange with the N- and C-terminal Ca2+ binding sites of calmodulin.
abstract: Myosin light chain kinase and peptides from the calmodulin (CaM) binding domains of myosin light chain kinase (RS-20, M-13), CaM kinase II, and the myristoylated alanine-rich protein kinase C substrate protein slowed Ca2+ dissociation from CaM's N-terminal sites from 405 +/- 75/s to 1.8-2.9/s and from CaM's C-terminal sites from 2.4 +/- 0.2/s to 0.1-0.4/s at 10 degrees C. Since Ca2+ dissociates 5-29 times faster from the N-terminal in these CaM.peptide complexes and both lobes are required for activation, Ca2+ dissociation from the N-terminal would control target protein inactivation. Ca2+ binds 70 times faster to the N-terminal (1.6 x 10(8) M-1 s-1) than the C-terminal sites (2.3 x 10(6) M-1 s-1). In a 0.6-ms half-width Ca2+ transient, Ca2+ occupied > 70% of the N-terminal but only 20% of the C-terminal sites. RS-20 produced a 9-fold and CaM kinase II a 6.3-fold increase in C-terminal Ca2+ affinity, suggesting that some target proteins may be bound to the C-terminal at resting [Ca2+]. When this is the case, Ca2+ exchange with the faster N-terminal sites may regulate CaM's activation and inactivation of these target proteins during a Ca2+ transient.
authors.affiliation: Department of Medical Biochemistry, Ohio University Medical Center, Columbus 43210, USA.
authors.forename: J D
authors.initials: JD
authors.lastname: Johnson
authors.forename: C
authors.initials: C
authors.lastname: Snyder
authors.forename: M
authors.initials: M
authors.lastname: Walsh
authors.forename: M
authors.initials: M
authors.lastname: Flynn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D003198:Computer Simulation; D008969:Molecular Sequence Data; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*
chemical_list: D002147:Calmodulin; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D009219:Myosin-Light-Chain Kinase; D002118:Calcium