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, T. R. Soderling, A. R. Means

    Research output: Contribution to journalArticle

    195 Scopus citations

    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 - Jan 1 1988

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

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  • Cite this

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