High-efficiency expression of rat protein kinase C-gamma in baculovirus-infected insect cells.
We have expressed rat protein kinase C-gamma in insect cells using a baculovirus vector. The yield of expressed protein kinase C-gamma is about 4% of total protein. The recombinant protein shows a prominent band at about 80 kDa on SDS-polyacrylamide gels, which can be identified as protein kinase C-gamma by Western blotting using monoclonal antibodies against protein kinase C-gamma. Upon incubation with [gamma-32P]ATP and in the presence of Ca2+, phosphatidylserine and diacylglycerol this protein autophosphorylates. Its enzyme activity shows the characteristic properties of mammalian protein kinase C.
Fiebich
B
B
University of Freiburg, Institute of Molecular Cell Biology, FRG.
Hug
H
H
Marmé
D
D
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002478:Cells, Cultured; D004075:Diglycerides / Q000494:pharmacology; D005822:Genetic Vectors; D066298:In Vitro Techniques; D007313:Insecta; D010718:Phosphatidylserines / Q000494:pharmacology; D010750:Phosphoproteins / Q000235:genetics; D010766:Phosphorylation; D011493:Protein Kinase C / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism
D004075:Diglycerides; D010718:Phosphatidylserines; D010750:Phosphoproteins; D011994:Recombinant Proteins; D011493:Protein Kinase C