Domain characterization of rabbit skeletal muscle myosin light chain kinase

B. Herring, James T. Stull, Patricia Gallagher

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Myosin light chain kinase can be divided into three distinct structural domains, an amino-terminal "tail," of unknown function, a central catalytic core and a carboxy-terminal calmodulin-binding regulatory region. We have used a combination of deletion mutagenesis and monoclonal antibody epitope mapping to define these domains more closely. A 2.95-kilobase cDNA has been isolated that includes the entire coding sequence of rabbit skeletal muscle myosin light chain kinase (607 amino acids). This cDNA, expressed in COS cells encoded a Ca2+/calmodulin-dependent myosin light chain kinase with a specific activity similar to that of the enzyme purified from rabbit skeletal muscle. Serial carboxy-terminal deletions of the regulatory and catalytic domains were constructed and expressed in COS cells. The truncated kinases had no detectable myosin light chain kinase activity. Monoclonal antibodies which inhibit the activity of the enzyme competitively with respect to myosin light chain were found to bind between residues 235-319 and 165-173, amino-terminal of the previously defined catalytic core. Thus, residues that are either involved in substrate binding or in close proximity to a light chain binding site may be located more amino-terminal than the previously defined catalytic core.

Original languageEnglish (US)
Pages (from-to)1724-1730
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number3
StatePublished - Jan 25 1990
Externally publishedYes

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Skeletal Muscle Myosins
Myosin-Light-Chain Kinase
Catalytic Domain
Rabbits
COS Cells
Calmodulin
Complementary DNA
Monoclonal Antibodies
Epitope Mapping
Myosin Light Chains
Mutagenesis
Nucleic Acid Regulatory Sequences
Enzymes
Muscle
Tail
Epitopes
Skeletal Muscle
Phosphotransferases
Binding Sites
Light

ASJC Scopus subject areas

  • Biochemistry

Cite this

Herring, B., Stull, J. T., & Gallagher, P. (1990). Domain characterization of rabbit skeletal muscle myosin light chain kinase. Journal of Biological Chemistry, 265(3), 1724-1730.

Domain characterization of rabbit skeletal muscle myosin light chain kinase. / Herring, B.; Stull, James T.; Gallagher, Patricia.

In: Journal of Biological Chemistry, Vol. 265, No. 3, 25.01.1990, p. 1724-1730.

Research output: Contribution to journalArticle

Herring, B, Stull, JT & Gallagher, P 1990, 'Domain characterization of rabbit skeletal muscle myosin light chain kinase', Journal of Biological Chemistry, vol. 265, no. 3, pp. 1724-1730.
Herring, B. ; Stull, James T. ; Gallagher, Patricia. / Domain characterization of rabbit skeletal muscle myosin light chain kinase. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 3. pp. 1724-1730.
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