Structure and function of smooth muscle myosin light chain kinase.
Myosin light chain kinase (MLCK) plays a central role in regulating the actin-myosin interaction of smooth muscle. MLCK phosphorylates the light chain of myosin in the presence of Ca2+ and calmodulin (CaM) thereby activating myosin so that it can interact with actin. Besides this kinase activity, MLCK shows i) actin-binding activity that can assemble actin filaments into their bundles and ii) myosin-binding activity that can form myosin filaments. To localize the actin- and myosin-binding activities in the MLCK molecule and to examine their possible role in regulating the actin-myosin interaction, we expressed various fragments of cDNA encoding MLCK in Escherichia coli as recombinant proteins. We found that MLCK consists of an N-terminal actin-binding domain, a central kinase domain, and a C-terminal myosin-binding domain. The Met1-Pro41 sequence is responsible for Ca2+/CaM-sensitive binding to actin. This binding site exerts an inhibitory effect on the actin-myosin interaction only when myosin is phosphorylated. MLCK binds to myosin at the C-terminal domain, the sequence of which is identical to telokin, an abundant myosin-binding protein in smooth muscle cells. This domain itself has no regulatory role in the interaction. However, the interaction was stimulated when this domain was extended to include the sequence known to regulate the activity of the kinase domain. The stimulation was observed only when myosin was unphosphorylated.
Kishi
H
H
Department of Pharmacology, Gunma University School of Medicine, Japan.
Ye
L H
LH
Nakamura
A
A
Okagaki
T
T
Iwata
A
A
Tanaka
T
T
Kohama
K
K
D000199:Actins / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D006801:Humans; D009130:Muscle, Smooth / Q000737:chemistry* / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000199:Actins; D002147:Calmodulin; D018076:DNA, Complementary; D011994:Recombinant Proteins; D009219:Myosin-Light-Chain Kinase; D002118:Calcium