title: Microbial transformation of steroids--IX. Purification of progesterone hydroxylase cytochrome P-450 from Phycomyces blakesleeanus.
abstract: Progesterone hydroxylase cytochrome P-450 was purified to homogeneity from Phycomyces blakesleeanus microsomes by a four step procedure. An M(r) value of 60,000 was determined for this protein by SDS-PAGE. The DEAE-cellulose and Blue-1 MIMETIC affinity fractions gave major peaks at 452 nm in a dithionite-reduced, carbon monoxide, difference spectrum. NaIO4-dependent progesterone hydroxylation was obtained by the pure enzyme without NADPH and NADPH-cytochrome P-450 reductase. NADPH-dependent hydroxylation required the addition of other Phycomyces microsomal proteins present in the Blue-1 fraction.
authors.affiliation: Department of Biochemistry, Queen Mary and Westfield College (University of London), U.K.
authors.forename: F
authors.initials: F
authors.lastname: Ahmed
authors.forename: R A
authors.initials: RA
authors.lastname: Williams
authors.forename: K E
authors.initials: KE
authors.lastname: Smith
mesh_terms: D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006900:Hydroxylation; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D010504:Periodic Acid / Q000378:metabolism; D010800:Phycomyces / Q000201:enzymology*; D011374:Progesterone / Q000378:metabolism*; D013250:Steroid Hydroxylases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D010504:Periodic Acid; D011374:Progesterone; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; C009288:metaperiodate; D013250:Steroid Hydroxylases