Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains

M. E. Payne, Y. L. Fong, T. Ono, R. J. Colbran, B. E. Kemp, Thomas Soderling, A. R. Means

Research output: Contribution to journalArticle

190 Citations (Scopus)

Abstract

Regulatory domains of the multifunctional Ca2+/calmodulin-dependent protein kinase II were investigated utilizing synthetic peptides. These peptides were derived from the sequence between positions 281 and 319 as translated from the cDNA sequence of the rat brain 50-kDa subunit (Lin, C.R., Kapiloff, M.S., Durgerian, S., Tatemoto, K., Russo, A.F., Hanson, P., Schulman, H., and Rosenfeld, M.G. (1987) Proc. Natl Acad. Sci. U.S.A. 84, 5962-5966), which contain the putative calmodulin-binding region as well as potential autophosphorylation sites. Peptide 290 to 309 was found to be a potent calmodulin antagonist with an IC50 of 52 nM for inhibition of Ca2+/calmodulin-dependent protein kinase II. Neither truncation from the amino terminus (peptide 296-309) nor extension in the carboxyl-terminal direction (peptide 294-319) markedly affected calmodulin binding, whereas shortening the peptide from the carboxyl terminus (peptide 290-302) or from both ends (peptide 295-304) resulted in the elimination of this activity. Peptide 281-290 did not bind calmodulin, but was a good substrate for the enzyme, being phosphorylated at Thr-286. Several of the peptides inhibited the kinase in a partially competitive, substrate-directed manner, but were not themselves phosphorylated. These studies identify domains within Ca2+/calmodulin-dependent protein kinase II which may be involved in 1) inhibition of the kinase in the absence of calmodulin, 2) binding of calmodulin, and 3) the resulting activation. Additionally, it is suggested that phosphorylation of residues flanking these domains may be responsible for the known regulatory effects of autophosphorylation on the properties of the kinase.

Original languageEnglish (US)
Pages (from-to)7190-7195
Number of pages6
JournalJournal of Biological Chemistry
Volume263
Issue number15
StatePublished - 1988
Externally publishedYes

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Calcium-Calmodulin-Dependent Protein Kinase Type 2
Calcium-Calmodulin-Dependent Protein Kinases
Calmodulin
Peptides
Phosphotransferases
Phosphorylation
Substrates
Inhibitory Concentration 50
Rats
Brain
Complementary DNA
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Payne, M. E., Fong, Y. L., Ono, T., Colbran, R. J., Kemp, B. E., Soderling, T., & Means, A. R. (1988). Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains. Journal of Biological Chemistry, 263(15), 7190-7195.

Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains. / Payne, M. E.; Fong, Y. L.; Ono, T.; Colbran, R. J.; Kemp, B. E.; Soderling, Thomas; Means, A. R.

In: Journal of Biological Chemistry, Vol. 263, No. 15, 1988, p. 7190-7195.

Research output: Contribution to journalArticle

Payne, ME, Fong, YL, Ono, T, Colbran, RJ, Kemp, BE, Soderling, T & Means, AR 1988, 'Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains', Journal of Biological Chemistry, vol. 263, no. 15, pp. 7190-7195.
Payne, M. E. ; Fong, Y. L. ; Ono, T. ; Colbran, R. J. ; Kemp, B. E. ; Soderling, Thomas ; Means, A. R. / Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains. In: Journal of Biological Chemistry. 1988 ; Vol. 263, No. 15. pp. 7190-7195.
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AU - Means, A. R.

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