Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities

D. P. Rich, R. J. Colbran, C. M. Schworer, Thomas Soderling

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Calcium/calmodulin (CaM)-dependent protein kinase II (CaM-kinase II) contained within the postsynaptic density (PSD) was shown to become partially Ca2+-independent following initial activation by Ca2+/CaM. Generation of this Ca2+-independent species was dependent upon autophosphorylation of both subunits of the enzyme in the presence of Mg2+/ATP/Ca2+/CaM and attained a maximal value of 74 ± 5% of the total activity within 1-2 min. Subsequent to the generation of this partially Ca2+-independent form of PSD CaM-kinase II, addition of EGTA to the autophosphorylation reaction resulted in further stimulation of 32PO4 incorporation into both kinase subunits and a loss of stimulation of the kinase by Ca2+/CaM. Examination of the sites of Ca2+-dependent autophosphorylation by phosphoamino acid analysis and peptide mapping of both kinase subunits suggested that phosphorylation of Thr(286/287) of the α and β-subunits, respectively, may be responsible for the transition of PSD CaM-kinase II to the Ca2+-independent species. A synthetic peptide 281-309 corresponding to a portion of the regulatory domain (residues 281-314) of the soluble kinase inhibited syntide-2 phosphorylation by the Ca2+-independent form of PSD CaM-kinase II (IC50 = 3.6 ± 0.8 μM). Binding of Ca2+/CaM to peptide 281-309 abolished its inhibitory property. Phosphorylation of Thr286 in peptide 281-309 also decreased its inhibitory potency. These data suggest that CaM-kinase II in the PSD possesses regulatory properties and mechanisms of activation similar to the cytosolic form of CaM-kinase II.

Original languageEnglish (US)
Pages (from-to)807-816
Number of pages10
JournalJournal of Neurochemistry
Volume53
Issue number3
StatePublished - 1989
Externally publishedYes

Fingerprint

Post-Synaptic Density
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Calcium-Calmodulin-Dependent Protein Kinases
Rats
Brain
Calmodulin
Phosphorylation
Phosphotransferases
Peptides
syntide-2
Chemical activation
Phosphoamino Acids
Peptide Mapping
Egtazic Acid
Inhibitory Concentration 50
Adenosine Triphosphate
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Rich, D. P., Colbran, R. J., Schworer, C. M., & Soderling, T. (1989). Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities. Journal of Neurochemistry, 53(3), 807-816.

Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities. / Rich, D. P.; Colbran, R. J.; Schworer, C. M.; Soderling, Thomas.

In: Journal of Neurochemistry, Vol. 53, No. 3, 1989, p. 807-816.

Research output: Contribution to journalArticle

Rich, DP, Colbran, RJ, Schworer, CM & Soderling, T 1989, 'Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities', Journal of Neurochemistry, vol. 53, no. 3, pp. 807-816.
Rich, D. P. ; Colbran, R. J. ; Schworer, C. M. ; Soderling, Thomas. / Regulatory properties of calcium/calmodulin-dependent protein kinase II in rat brain postsynaptic densities. In: Journal of Neurochemistry. 1989 ; Vol. 53, No. 3. pp. 807-816.
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