TY - JOUR
T1 - Expression and characterization of the alpha-subunit of Ca2+/calmodulin-dependent protein kinase II using the baculovirus expression system
AU - Brickey, Debra A.
AU - Colbran, Roger J.
AU - Fong, Yiu Lian
AU - Soderling, Thomas R.
N1 - Funding Information:
Acknowledgments--We would like to thank Martha Bass and Elizabeth Wolf for their excellent technical assistance. This work was supported by National Institutes of Health Grants GM41292 and DK17808.
PY - 1990/12/14
Y1 - 1990/12/14
N2 - 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.
AB - 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.
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U2 - 10.1016/S0006-291X(05)80074-9
DO - 10.1016/S0006-291X(05)80074-9
M3 - Article
C2 - 2175600
AN - SCOPUS:0025666672
SN - 0006-291X
VL - 173
SP - 578
EP - 584
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -