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 language||English (US)|
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Dec 14 1990|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology