Activation mechanisms for Ca2+/calmodulin-dependent protein kinase IV: Identification of a brain CaM-kinase IV kinase

Hiroshi Tokumitsu, Debra A. Brickey, John Glod, Hiroyoshi Hidaka, James Sikela, Thomas R. Soderling

Research output: Contribution to journalArticle

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Abstract

This manuscript examines the mechanisms by which Ca2Vcalmodulin-dependent protein kinase IV (CaM-kinase IV) is activated through the binding of Ca2+/CaM and by phosphorylation. Studies with the synthetic autoinhibitory domain peptides of CaM-kinase II indicate that CaM-kinase IV has a similarly located autoinhibitory domain, and this was confirmed since site-directed mutagenesis of this region (HMDT308 to DEDD and FN317 to DD) generated fully active Ca2+/CaM-independent kinases. Total activities of purified, baculovirus-expressed wild type and mutant kinases were increased 2-fold by intramolecular autophosphorylation, but this reaction was extremely slow (1-2 h) and probably not physiological. However, CaM-kinase FV can be activated by brain CaM-kinase FV kinase resulting in large increases in both total (5-7-fold) and Ca2+/CaM-independent (>20-fold) CaM-kinase IV activities. This activation reaction required Mg2+/ATP and Ca2+/CaM, was intermolecularly catalyzed, and was reversed by protein phosphatase 2A. Activation of CaM-kinase IV resulted in a 10-fold decrease in Km for syntide-2 with little effect on Km for ATP or Vmax. CaM-kinase FV kinase was highly purified from rat brain extract and was shown to be a 68-kDa monomer. The results of this study demonstrate that CaM-kinase IV does have an autoinhibitory domain within residues His305-Lys321 that suppresses kinase activity in the absence of Ca2+/CaM. CaM-kinase IV is not significantly activated by autophosphorylation, but it can be activated 10-fold by a CaM-kinase IV kinase. This kinase cascade activation mechanism may be important for the physiological function of CaM-kinase IV such as transcriptional regulation through phosphorylation of cAMP responsive element binding protein (Enslen, H., Sun, P., Brickey, D., Soderling, S. H., Klamo, E., and Soderling, T. R. (1994) J. Biol. Chem. 269, 15520-15527).

Original languageEnglish
Pages (from-to)28640-28647
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number46
Publication statusPublished - Nov 18 1994
Externally publishedYes

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Calcium-Calmodulin-Dependent Protein Kinase Type 4
Protein Kinases
Brain
Phosphotransferases
Chemical activation
Phosphorylation
syntide-2
Adenosine Triphosphate
Protein Phosphatase 2
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Mutagenesis
Baculoviridae
Site-Directed Mutagenesis
Rats
Carrier Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activation mechanisms for Ca2+/calmodulin-dependent protein kinase IV : Identification of a brain CaM-kinase IV kinase. / Tokumitsu, Hiroshi; Brickey, Debra A.; Glod, John; Hidaka, Hiroyoshi; Sikela, James; Soderling, Thomas R.

In: Journal of Biological Chemistry, Vol. 269, No. 46, 18.11.1994, p. 28640-28647.

Research output: Contribution to journalArticle

Tokumitsu, H, Brickey, DA, Glod, J, Hidaka, H, Sikela, J & Soderling, TR 1994, 'Activation mechanisms for Ca2+/calmodulin-dependent protein kinase IV: Identification of a brain CaM-kinase IV kinase', Journal of Biological Chemistry, vol. 269, no. 46, pp. 28640-28647.
Tokumitsu, Hiroshi ; Brickey, Debra A. ; Glod, John ; Hidaka, Hiroyoshi ; Sikela, James ; Soderling, Thomas R. / Activation mechanisms for Ca2+/calmodulin-dependent protein kinase IV : Identification of a brain CaM-kinase IV kinase. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 46. pp. 28640-28647.
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AU - Soderling, Thomas R.

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