Phosphorylation of synthetic peptides by skeletal muscle myosin light chain kinases

C. H. Michnoff, B. E. Kemp, J. T. Stull

Research output: Contribution to journalArticle

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Abstract

Substrate determinants for rabbit and chicken skeletal muscle myosin light chain kinases were examined with synthetic peptides. Both skeletal muscle myosin light chain kinases had similar phosphorylation kinetics with synthetic peptide substrates. Average kinetic constants for skeletal muscle myosin light chain heptadecapeptide, P-K-K-A-K-R-R-A-A-E-G-S-S(P)-N-V-F-S where S(P) is phosphoserine, were K(m), 2.3 μM and V(max), 0.9 μmol/min/mg of enzyme. K(m) values were 122 and 162 μM for skeletal muscle peptides containing A-A for basic residues at positions 2-3 and 6-7, respectively. Average kinetic constants for smooth muscle myosin light chain peptide, S-S-K-R-A-K-A-K-T-T-K-K-R-P-Q-R-A-T-S(P)-N-V-F-S, were K(m), 1.4 μM and V(max), 27 μmol/min/mg of enzyme. Average K(m) values for the smooth muscle peptide, residues 11-23, were 10 μM which increased 6- and 11-fold with substitutions of alanine at residues 12 and 13, respectively. V(max) values decreased and K(m) values increased markedly by substitution of residue 16 with glutamate in the 11-23 smooth muscle tridecapeptide. Basic residues located 3 and 6-7 residues toward the NH2 terminus from phosphoserine in smooth muscle myosin light chain and 6-8 and 10-11 residues toward the NH2 terminus from phosphoserin in skeletal muscle myosin light chain appear to be important substrate determinants for skeletal muscle myosin light chain kinases. These properties are different from myosin light chain kinase from smooth muscle.

Original languageEnglish (US)
Pages (from-to)8320-8326
Number of pages7
JournalJournal of Biological Chemistry
Volume261
Issue number18
StatePublished - 1986

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Skeletal Muscle Myosins
Myosin-Light-Chain Kinase
Phosphorylation
Myosin Light Chains
Muscle
Peptides
Smooth Muscle Myosins
Smooth Muscle
Phosphoserine
Kinetics
Substitution reactions
Substrates
Enzymes
Alanine
Glutamic Acid
Chickens
Skeletal Muscle
Rabbits

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of synthetic peptides by skeletal muscle myosin light chain kinases. / Michnoff, C. H.; Kemp, B. E.; Stull, J. T.

In: Journal of Biological Chemistry, Vol. 261, No. 18, 1986, p. 8320-8326.

Research output: Contribution to journalArticle

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N2 - Substrate determinants for rabbit and chicken skeletal muscle myosin light chain kinases were examined with synthetic peptides. Both skeletal muscle myosin light chain kinases had similar phosphorylation kinetics with synthetic peptide substrates. Average kinetic constants for skeletal muscle myosin light chain heptadecapeptide, P-K-K-A-K-R-R-A-A-E-G-S-S(P)-N-V-F-S where S(P) is phosphoserine, were K(m), 2.3 μM and V(max), 0.9 μmol/min/mg of enzyme. K(m) values were 122 and 162 μM for skeletal muscle peptides containing A-A for basic residues at positions 2-3 and 6-7, respectively. Average kinetic constants for smooth muscle myosin light chain peptide, S-S-K-R-A-K-A-K-T-T-K-K-R-P-Q-R-A-T-S(P)-N-V-F-S, were K(m), 1.4 μM and V(max), 27 μmol/min/mg of enzyme. Average K(m) values for the smooth muscle peptide, residues 11-23, were 10 μM which increased 6- and 11-fold with substitutions of alanine at residues 12 and 13, respectively. V(max) values decreased and K(m) values increased markedly by substitution of residue 16 with glutamate in the 11-23 smooth muscle tridecapeptide. Basic residues located 3 and 6-7 residues toward the NH2 terminus from phosphoserine in smooth muscle myosin light chain and 6-8 and 10-11 residues toward the NH2 terminus from phosphoserin in skeletal muscle myosin light chain appear to be important substrate determinants for skeletal muscle myosin light chain kinases. These properties are different from myosin light chain kinase from smooth muscle.

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