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The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence.

Publication ,  Journal Article
Kemp, BE; Pearson, RB; Guerriero, V; Bagchi, IC; Means, AR
Published in: J Biol Chem
February 25, 1987

Smooth muscle myosin light chain kinase contains a 64 residue sequence that binds calmodulin in a Ca2+-dependent manner (Guerriero, V., Jr., Russo, M. A., and Means, A. R. (1987) Biochemistry, in press). Within this region is a sequence with homology to the corresponding sequence reported for the calmodulin binding region of skeletal muscle myosin light chain kinase (Blumenthal, D. K., Takio, K., Edelman, A. M., Charbonneau, H., Titani, L., Walsh, K. A., and Krebs, E. G. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 3187-3191). Inspection of these sequences reveals that they both share a similar number and spatial arrangement of basic residues with those present in the myosin light chain substrate. We have synthesized a 22-residue peptide corresponding to residues 480-501 (determined from the cDNA) of the smooth muscle myosin light chain kinase. This peptide, Ala-Lys-Lys-Leu-Ser-Lys-Asp-Arg-Met-Lys-Lys-Tyr-Met-Ala-Arg-Arg-Lys-Trp- Gln-Lys-Thr-Gly, inhibited calmodulin-dependent activation of the smooth muscle myosin light chain kinase with an IC50 of 46 nM. At saturating concentrations of calmodulin, the 22-residue peptide inhibited myosin light chain and synthetic peptide substrate phosphorylation competitively with IC50 values of 2.7 and 0.9 microM, respectively. An 11-residue synthetic peptide analog, corresponding to part of the calmodulin-binding sequence in skeletal muscle myosin light chain kinase, Lys-Arg-Arg-Trp-Lys-Lys-Asn-Phe-Ile-Ala-Val, also competitively inhibited synthetic peptide substrate phosphorylation with a Ki of 1 microM. The competitive inhibitory activity of the calmodulin binding regions is similar to the apparent Km of 2.7 microM for phosphorylation of the 23-residue peptide analog of the smooth muscle myosin light chain and raises the possibility that the calmodulin binding region of the myosin light chain kinase may act as a pseudosubstrate inhibitor of the enzyme.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

February 25, 1987

Volume

262

Issue

6

Start / End Page

2542 / 2548

Location

United States

Related Subject Headings

  • Phosphorylation
  • Myosin-Light-Chain Kinase
  • Muscles
  • Muscle, Smooth
  • Models, Chemical
  • Kinetics
  • Chickens
  • Calmodulin
  • Biochemistry & Molecular Biology
  • Binding, Competitive
 

Citation

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Kemp, B. E., Pearson, R. B., Guerriero, V., Bagchi, I. C., & Means, A. R. (1987). The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence. J Biol Chem, 262(6), 2542–2548.
Kemp, B. E., R. B. Pearson, V. Guerriero, I. C. Bagchi, and A. R. Means. “The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence.J Biol Chem 262, no. 6 (February 25, 1987): 2542–48.
Kemp BE, Pearson RB, Guerriero V, Bagchi IC, Means AR. The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence. J Biol Chem. 1987 Feb 25;262(6):2542–8.
Kemp, B. E., et al. “The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence.J Biol Chem, vol. 262, no. 6, Feb. 1987, pp. 2542–48.
Kemp BE, Pearson RB, Guerriero V, Bagchi IC, Means AR. The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence. J Biol Chem. 1987 Feb 25;262(6):2542–2548.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

February 25, 1987

Volume

262

Issue

6

Start / End Page

2542 / 2548

Location

United States

Related Subject Headings

  • Phosphorylation
  • Myosin-Light-Chain Kinase
  • Muscles
  • Muscle, Smooth
  • Models, Chemical
  • Kinetics
  • Chickens
  • Calmodulin
  • Biochemistry & Molecular Biology
  • Binding, Competitive