Expression of a multifunctional Ca2+/calmodulin-dependent protein kinase and mutational analysis of its autoregulation

Phyllis I. Hanson, Michael S Kapiloff, Lillian L. Lou, Michael G. Rosenfeld, Howard Schulman

Research output: Contribution to journalArticle

220 Citations (Scopus)

Abstract

Autophosphorylation of multifunctional Ca2+/calmod-ulin-dependent protein kinase converts it from a Ca2+-dependent to a Ca2+-independent or autonomous kinase, a process that may underlie some long-term enhancement of transient Ca2+ signals. We demonstrate that the neuronal α subunit clone expressed in COS-7 cells (α-CaM kinase) is sufficient to encode the regulatory phenomena characteristic of the multisubunit kinase isolated from brain. Activity of α-CaM kinase is highly dependent on Ca2+/calmodulin. It is converted by autophosphorylation to an enzyme capable of Ca2+-independent (autonomous) substrate phosphorylation and autophosphorylation. Using site-directed mutagenesis, we separately eliminate five putative auto-phosphorylation sites within the regulatory domain and directly examine their individual roles. Ca2+/calmodulin-dependent kinase activity is fully retained by each mutant, but Thr286 is unique among the sites in being indispensable for generation of an autonomous kinase.

Original languageEnglish
Pages (from-to)59-70
Number of pages12
JournalNeuron
Volume3
Issue number1
DOIs
StatePublished - Jan 1 1989
Externally publishedYes

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Calcium-Calmodulin-Dependent Protein Kinases
Homeostasis
Phosphotransferases
Phosphorylation
COS Cells
Calmodulin
Site-Directed Mutagenesis
Protein Kinases
Clone Cells
Brain
Enzymes

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Expression of a multifunctional Ca2+/calmodulin-dependent protein kinase and mutational analysis of its autoregulation. / Hanson, Phyllis I.; Kapiloff, Michael S; Lou, Lillian L.; Rosenfeld, Michael G.; Schulman, Howard.

In: Neuron, Vol. 3, No. 1, 01.01.1989, p. 59-70.

Research output: Contribution to journalArticle

Hanson, Phyllis I. ; Kapiloff, Michael S ; Lou, Lillian L. ; Rosenfeld, Michael G. ; Schulman, Howard. / Expression of a multifunctional Ca2+/calmodulin-dependent protein kinase and mutational analysis of its autoregulation. In: Neuron. 1989 ; Vol. 3, No. 1. pp. 59-70.
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