Isolation of the cDNA encoding rat skeletal muscle myosin light chain kinase. Sequence and tissue distribution.
A cDNA clone encoding skeletal muscle myosin light chain kinase (MLCK) was isolated from a rat skeletal muscle library using oligonucleotide probes. The total length of the rat skeletal muscle MLCK cDNA was 2823 base pairs with an open reading frame of 1830 base pairs. The deduced sequence of the 610-amino acid protein exhibited 96% amino acid identity to rabbit skeletal muscle MLCK in the carboxyl-terminal portion of the molecule, which contains the catalytic and the calmodulin-binding domains, and 58% identity in the amino-terminal region. Analysis of total rat mRNA revealed a single mRNA species of 3.4 kilobases that was unique to skeletal muscle. Further analysis of skeletal muscle tissue using fast-twitch glycolytic, fast-twitch oxidative glycolytic, and slow-twitch oxidative fibers isolated from rat leg revealed that the mRNA level for MLCK varied among the three fiber types. The results of kinase assays performed on the fibers showed that MLCK activity levels paralleled the MLCK mRNA levels found in each of the three types of skeletal muscle fibers studied. Fast-twitch oxidative glycolytic (gastrocnemius red) and slow-twitch oxidative (soleus) exhibited 60 and 13%, respectively, of the enzymatic activity present in fast-twitch glycolytic (gastrocnemius white) fibers.
Roush
C L
CL
Department of Pharmacology, Howard Hughes Medical Institute, University of Washington, Seattle 98195.
Kennelly
P J
PJ
Glaccum
M B
MB
Helfman
D M
DM
Scott
J D
JD
Krebs
E G
EG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D005796:Genes; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology*; D009219:Myosin-Light-Chain Kinase / Q000235:genetics*; D009710:Nucleotide Mapping; D009928:Organ Specificity; D011817:Rabbits; D051381:Rats; D013045:Species Specificity
D004247:DNA; D009219:Myosin-Light-Chain Kinase; D004262:DNA Restriction Enzymes