Expression and transport into mitochondria of bovine cytochrome P-450(SCC) in insect cells using the baculovirus expression system

Biochem Biophys Res Commun. 1992 Apr 15;184(1):471-7. doi: 10.1016/0006-291x(92)91218-f.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cattle
  • Cell Line
  • Cholesterol Side-Chain Cleavage Enzyme / biosynthesis
  • Cholesterol Side-Chain Cleavage Enzyme / genetics*
  • Cholesterol Side-Chain Cleavage Enzyme / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression
  • Genetic Techniques
  • Genetic Vectors
  • Kinetics
  • Mitochondria / enzymology*
  • Moths
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Transfection*
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics
  • beta-Galactosidase / isolation & purification

Substances

  • Recombinant Fusion Proteins
  • Cholesterol Side-Chain Cleavage Enzyme
  • beta-Galactosidase