Expression and transport into mitochondria of bovine cytochrome P-450(SCC) in insect cells using the baculovirus expression system.
Bovine cytochrome P-450(SCC) introduced with the baculovirus host vector system was found to be expressed in Spodoptera frugiperda cells. Cell fractionation analysis indicated that the P-450(SCC) expressed as the precursor form was transported into mitochondria and converted to a mature form. However, this form did not exhibit definite activity for cholesterol side chain cleavage. These findings suggest that most of the P-450(SCC) expressed by this system is an inactive protein within mitochondria that is not folded to the conformation of the active enzyme and/or does not incorporate heme appropriately.
Takagi
Y
Y
Department of Biochemistry, School of Medicine, Keio University, Tokyo, Japan.
Ogawa
H
H
Harada
N
N
Shimada
H
H
Ishimura
Y
Y
Takagi
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002417:Cattle; D002460:Cell Line; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005821:Genetic Techniques; D005822:Genetic Vectors; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology*; D009036:Moths; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D002786:Cholesterol Side-Chain Cleavage Enzyme; D001616:beta-Galactosidase