title: A molecular mechanism for autoinhibition of myosin light chain kinases.
abstract: It is postulated that basic residues within the inhibitory region of myosin light chain kinase (MLCK) bind acidic residues within the catalytic core to maintain the kinase in an inactive form. In this study, we identified residues within the catalytic cores of the skeletal and smooth muscle MLCKs that may bind basic residues in inhibitory region. Acidic residues within the catalytic core of the rabbit skeletal and smooth muscle MLCKs were mutated and the kinetic properties of the mutant kinases determined. Mutation of 6 and 8 acidic residues in the skeletal and smooth muscle MLCKs, respectively, result in mutant MLCKs with decreases in KCaM (the concentration of calmodulin required for half-maximal activation of myosin light chain kinase) value ranging from 2- to 100-fold. Two inhibitory domain binding residues identified in each kinase also bind a basic residue in light chain substrate. The remaining mutants all have wild-type Km values for light chain. The predicted inhibitory domain binding residues are distributed in a linear fashion across the surface of the lower lobe of the proposed molecular model of the smooth muscle MLCK catalytic core. As 6 of the inhibitory domain binding residues in the smooth muscle MLCK are conserved in other Ca2+/calmodulin-dependent protein kinases, the structural basis for autoinhibition and activation may be similar.
authors.affiliation: Department of Physiology, University of Texas Southwestern Medical Center, Dallas 75235-9040.
authors.forename: P J
authors.initials: PJ
authors.lastname: Gallagher
authors.forename: B P
authors.initials: BP
authors.lastname: Herring
authors.forename: A
authors.initials: A
authors.lastname: Trafny
authors.forename: J
authors.initials: J
authors.lastname: Sowadski
authors.forename: J T
authors.initials: JT
authors.lastname: Stull
mesh_terms: D000818:Animals; D002384:Catalysis; D002417:Cattle; D002460:Cell Line; D008958:Models, Molecular; D009130:Muscle, Smooth / Q000201:enzymology; D009132:Muscles / Q000201:enzymology; D009154:Mutation; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011817:Rabbits
chemical_list: D009219:Myosin-Light-Chain Kinase