Biochemical properties of chimeric skeletal and smooth muscle myosin light chain kinases.
The molecular and biochemical properties of myosin light chain kinases from chicken skeletal and smooth muscle were investigated by recombinant DNA techniques. Deletion of the amino-terminal region of either the smooth or skeletal muscle myosin light chain kinase resulted in a decrease in Vmax with no significant change in Km values for light chain substrates. Skeletal/smooth muscle chimeric kinases were inactive when a 65-residue region amino-terminal of the catalytic core was exchanged between the two forms. Changing alanine 494 to glutamic acid within this region in the chicken skeletal muscle myosin light chain kinase increased the Km values for light chains 10-fold. These results are consistent with the hypothesis that the region amino-terminal of the catalytic core in myosin light chain kinases is involved in light chain recognition. A skeletal muscle kinase which contained the smooth muscle calmodulin binding domain remained regulated by Ca2+/calmodulin. Thus, the calmodulin binding domains of smooth and skeletal muscle myosin light chain kinases share structural elements necessary for regulation.
Department of Physiology, University of Texas Southwestern Medical Center, Dallas 75235.
S A
SA
Leachman
P J
PJ
Gallagher
B P
BP
Herring
M J
MJ
McPhaul
J T
JT
Stull
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002147:Calmodulin / Q000378:metabolism; D002645:Chickens; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000201:enzymology*; D009132:Muscles / Q000201:enzymology*; D009219:Myosin-Light-Chain Kinase / Q000235:genetics / Q000378:metabolism*; D012313:RNA / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D014158:Transcription, Genetic
D002147:Calmodulin; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D012313:RNA; D004247:DNA; D009219:Myosin-Light-Chain Kinase