Ca(2+)-calmodulin-dependent modification of smooth-muscle myosin light-chain kinase leading to its co-operative activation by calmodulin.
It has recently been shown that at relatively high molar ratios of myosin light-chain kinase (MLCKase) to calmodulin (CM) almost complete inhibition of the kinase activity occurs [Sobieszek (1991) J. Mol. Biol. 220, 947-957]. This inhibition resulted in a highly co-operative activation of MLCKase by CM, whereas the opposite activation (of CM by kinase) was hyperbolic, as expected (unco-operative). This difference in activation was observed only for kinase preparations preincubated with sub-stoichiometric amounts of CM, and only when micromolar concentrations of Ca2+ were present. The inhibitory effect was variable and depended not only on the concentration ratio of kinase to CM but also on the MLCKase preparation. For most of the preparations full inhibition required 5-15 min of preincubation at 25 degrees C and a 3-6-fold molar excess of kinase over CM. The inhibition was reversible, since full activity could be obtained after saturation of the kinase by additional CM. The inhibitory effect did not require ATP (excluding phosphorylation-type modifications of the kinase), and dephosphorylation of the kinase was not involved, since inhibition of an endogenous MLCK phosphatase by microcystin-LR did not decrease the inhibitory effect. Since the co-operative activation by CM was observed for cross-linked MLCKase preparations enriched in kinase dimers, but was absent for the analogous preparations enriched in the oligomers, we concluded that Ca(2+)-CM-dependent changes in the oligomeric state of the kinase were responsible for the modification observed. The exact nature of these modifications remains to be established.
Sobieszek
A
A
Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg.
Strobl
A
A
Ortner
B
B
Babiychuk
E B
EB
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D004789:Enzyme Activation; D009130:Muscle, Smooth / Q000201:enzymology*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D043263:Myosin-Light-Chain Phosphatase; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D013552:Swine; D014422:Turkeys
D002147:Calmodulin; D000255:Adenosine Triphosphate; D009219:Myosin-Light-Chain Kinase; D010749:Phosphoprotein Phosphatases; D043263:Myosin-Light-Chain Phosphatase; D002118:Calcium