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 R. Soderling

    Research output: Contribution to journalArticle

    62 Scopus citations

    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

    ASJC Scopus subject areas

    • Biophysics
    • Biochemistry
    • Molecular Biology
    • Cell Biology

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