Contribution of cytochrome b5 to androgen synthesis in rat testicular microsomes.
Cytochrome b5 was purified from porcine testicular microsomes, and its amino acid composition was determined. Rabbit antibody against the purified cytochrome b5 was prepared in order to study the contribution of cytochrome b5 to testicular microsomal oxygenases related to androgen production. In the presence of NADPH alone as the electron donor, the antibody against cytochrome b5 inhibited the activities of steroid 17 alpha-hydroxylase and C-17-C-20 lyase of rat testicular microsomal fraction. Addition of NADH to the NADPH-supported oxygenase assay system enhanced both steroid oxygenase activities, and addition of the antibody against cytochrome b5 decreased the NADH-caused stimulation of steroid 17 alpha-hydroxylase and C-17-C-20 lyase activities. When dehydroepiandrosterone and NAD+ were added as substrates for 3 beta-hydroxy-delta 5-steroid dehydrogenase in order to synthesize NADH by enzymatic reaction, the NADPH-supported activities of steroid 17 alpha-hydroxylase and C-17-C-20 lyase were further stimulated as compared with the addition of NADH, and this stimulation was suppressed by the antibody against cytochrome b5. These results suggest that cytochrome b5, together with 3 beta-hydroxy-delta 5-steroid dehydrogenase, contributes to the activities of steroid 17 alpha-hydroxylase and C-17-C-20 lyase in the testicular microsomal fraction.
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000446:Aldehyde-Lyases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000728:Androgens / Q000096:biosynthesis*; D000818:Animals; D003573:Cytochrome b Group / Q000032:analysis / Q000378:metabolism*; D015786:Cytochromes b5; D004579:Electron Transport; D007118:Immunoassay; D005779:Immunodiffusion; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D008297:Male; D008861:Microsomes / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism; D013552:Swine; D013737:Testis / Q000378:metabolism*
D000596:Amino Acids; D000728:Androgens; D003573:Cytochrome b Group; D015786:Cytochromes b5; D015096:3-Hydroxysteroid Dehydrogenases; D013254:Steroid 17-alpha-Hydroxylase; D000446:Aldehyde-Lyases