Expression and characterization of the alpha-subunit of Ca2+/calmodulin-dependent protein kinase II using the baculovirus expression system

Debra A. Brickey, Roger J. Colbran, Yiu Lian Fong, Thomas Soderling

Research output: Contribution to journalArticle

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Abstract

Sf9 cells infected with the recombinant mouse CaMKII-α (Ca2+/calmodulin dependent kinase II) baculovirus expressed 12-15 mg of MCaMKII-α per liter of cells. Approximately 50% of the MCaMKII-α activity could be purified using a CaM-Sepharose affinity column. The purified MCaMKII-α had a Mrapp of 50 kDa by SDS-PAGE and a native Mr of 600 kDa. MCaMKII-α, like rat brain CaMKII, had an A0.5 for CaM of 100 nM, a Km for syntide-2 of 8 μM, and was able to generate Ca2+-independent activity by autophosphorylation. The baculovirus system expressed large quantites of MCaMKII-α with characteristics similar to the rat brain CaMKII, thus providing an expression system for the detailed biochemical analysis of MCaMKII-α.

Original languageEnglish (US)
Pages (from-to)578-584
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume173
Issue number2
DOIs
StatePublished - Dec 14 1990
Externally publishedYes

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Calcium-Calmodulin-Dependent Protein Kinase Type 2
syntide-2
Baculoviridae
Rats
Brain
Sf9 Cells
Calcium-Calmodulin-Dependent Protein Kinases
Calmodulin
Sepharose
Polyacrylamide Gel Electrophoresis
Phosphotransferases

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Expression and characterization of the alpha-subunit of Ca2+/calmodulin-dependent protein kinase II using the baculovirus expression system. / Brickey, Debra A.; Colbran, Roger J.; Fong, Yiu Lian; Soderling, Thomas.

In: Biochemical and Biophysical Research Communications, Vol. 173, No. 2, 14.12.1990, p. 578-584.

Research output: Contribution to journalArticle

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