[Immunochemical study of cholesterol-hydroxylating cytochrome P-450. Topology of the peptide chain of the heme protein in the phospholipid membrane].
The topology of adrenocortical cytochrome P-450scc in inner mitochondrial membrane was studied. To determine the polypeptide chain parts exposed to matrix or cytosol, two approaches were used, i.e. i) limited proteolysis of membrane-bound cytochrome P-450scc followed by the detection of the peptides formed by immunoblotting; ii) binding of monospecific antibodies against cytochrome P-450scc as well as fragments F1 and F2 representing N- and C-terminal sequences of the hemeprotein, to membrane structures (mitoplasts and submitochondrial particles). The data obtained confirm the transmembrane orientation of cytochrome P-450scc molecule, since antibodies against the hemeprotein as well as fragments F1 and F2 were found to be bound both on the matrix and cytosol surfaces of the inner mitochondrial membrane. It was shown that region 250-257 in cytochrome P-450scc connecting domains F1 and F2 is exposed to the matrix. A model of molecular organization of cytochrome P-450scc in inner mitochondrial membranes is proposed.
Usanov
S A
SA
Chernogolov
A A
AA
Chashchin
V L
VL
D000302:Adrenal Cortex / Q000201:enzymology; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002790:Cholesterol 7-alpha-Hydroxylase / Q000032:analysis*; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006420:Hemeproteins / Q000032:analysis*; D006868:Hydrolysis; D008563:Membrane Lipids / Q000032:analysis*; D008854:Microscopy, Electron; D008928:Mitochondria / Q000201:enzymology; D010743:Phospholipids / Q000032:analysis*; D011487:Protein Conformation; D011863:Radioimmunoassay; D013250:Steroid Hydroxylases / Q000032:analysis*; D013367:Submitochondrial Particles / Q000201:enzymology
D006420:Hemeproteins; D008563:Membrane Lipids; D010743:Phospholipids; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D002790:Cholesterol 7-alpha-Hydroxylase
Immunokhimicheskoe issledovanie kholesteringidroksiliruiushchego tsitokhroma P-450. Topologiia polipeptidnoĭ tsepi gemoproteida v fosfolipidnoĭ membrane.