[Immunochemical characteristics of cholesterol-hydroxylating cytochrome P-450. Monospecific antibodies against domains F1 and F2].
Highly specific antibodies to adrenocortical cytochrome P-450scc as well as fragments F1 and F2 representing the N- and C-terminal sequences of the hemoprotein obtained by limited trypsinolysis were raised in rabbits. Antibodies to cytochrome P-450scc as demonstrated by the Ouchterlony diffusion analysis, immunoelectrophoresis and immunoblotting techniques interact with the hemoprotein and both fragments. Antibodies to cytochrome P-450scc fragments interact with the hemoprotein and corresponding antigens, but do not cross-react. To determine the localization of antigenic determinants in the polypeptide chain of cytochrome P-450scc, the interaction of antibodies to the hemoprotein and to its fragments F1 and F2 with limited trypsinolysis products was studied. All antibodies were found to effectively inhibit cholesterol transformation into pregnenolone in a reconstituted system. Using SDS electrophoresis followed by immunoblotting, the cross-reactivity of antibodies to cytochrome P-450scc and to its fragments with microsomal cytochromes P-450scc LM2 and LM4 as well as with mitochondrial cytochrome P-45027 was revealed. This finding testifies to the presence of common antigenic determinants in the hemoproteins.
D000302:Adrenal Cortex / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002784:Cholesterol / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000276:immunology; D000939:Epitopes / Q000032:analysis; D007120:Immunochemistry; D008862:Microsomes, Liver / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D011817:Rabbits
D000939:Epitopes; D003577:Cytochrome P-450 Enzyme System; D002784:Cholesterol
Immunokhimicheskaia kharakteristika kholesteringidroksiliruiushchego tsitokhroma P-450. Monospetsificheskie antitela k domenam F1 i F2.