The role of porcine cytochrome b5A and cytochrome b5B in the regulation of cytochrome P45017A1 activities.
Male pigs are routinely castrated to prevent the accumulation of testicular 16-androstene steroids, in particular 5alpha-androst-16-en-3-one (5alpha-androstenone), which contribute to an off-odour and off-flavour known as boar taint. Cytochrome P450C17 (CYP17A1) catalyses the key regulatory step in the formation of the 16-androstene steroids from pregnenolone by the andien-beta synthase reaction or the synthesis of the glucocorticoid and sex steroids via 17alpha-hydroxylase and C17,20 lyase pathways respectively. We have expressed CYP17A1, along with cytochrome P450 reductase (POR), cytochrome b5 reductase (CYB5R3) and cytochrome b5 (CYB5) in HEK-293FT cells to investigate the importance of the two forms of porcine CYB5, CYB5A and CYB5B, in both the andien-beta synthase as well as the 17alpha-hydroxylase and C17,20 lyase reactions. Increasing the ratio of CYB5A to CYP17A1 caused a decrease in 17alpha-hydroxylase (p<0.013), a transient increase in C17,20 lyase, and an increase in andien-beta synthase activity (p<0.0001). Increasing the ratio of CYB5B to CYP17A1 also decreased 17alpha-hydroxylase, but did not affect the andien-beta synthase activity; however, the C17,20 lyase, was significantly increased. These results demonstrate the differential effects of two forms of CYB5 on the three activities of porcine CYP17A1 and show that CYB5B does not stimulate the andien-beta synthase activity of CYP17A1.
Billen
M J
MJ
Department of Animal Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Squires
E J
EJ
D006907:17-alpha-Hydroxypregnenolone / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D015786:Cytochromes b5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003687:Dehydroepiandrosterone / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000378:metabolism; D020033:Protein Isoforms / Q000378:metabolism; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism*; D034421:Sus scrofa / Q000378:metabolism*
D020033:Protein Isoforms; D006907:17-alpha-Hydroxypregnenolone; D003687:Dehydroepiandrosterone; D015786:Cytochromes b5; D010088:Oxidoreductases; C019701:androsta-5,16-dien-3 beta-ol synthetase; D013254:Steroid 17-alpha-Hydroxylase