title: Purification of cytochrome P-450 from adult pig testis by hydroxylapatite and deoxycorticosterone affinity column chromatography.
abstract: Adult testicular cytochrome P-450 was purified by a two-step procedure utilizing hydroxylapatite and deoxycorticosterone affinity column chromatography. Cytochrome P-450 was determined to have an isoelectric point of 6.5 on analytical isoelectric focusing. The purified cytochrome P-450 was found to be homogeneous and its molecular mass was estimated to be 52000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The carbon monoxide difference spectrum with a peak at 448 nm exhibited the absorption spectrum of a typical cytochrome P-450. A 1000-fold purification was achieved with a yield of 5%.
authors.affiliation: Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara, Kanagawa, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kuwada
authors.forename: J
authors.initials: J
authors.lastname: Maki
authors.forename: H
authors.initials: H
authors.lastname: Hasumi
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D003900:Desoxycorticosterone / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D006882:Hydroxyapatites / Q000737:chemistry; D007525:Isoelectric Focusing; D008297:Male; D008861:Microsomes / Q000201:enzymology; D013552:Swine; D013737:Testis / Q000201:enzymology*
chemical_list: D006882:Hydroxyapatites; D003900:Desoxycorticosterone; D003577:Cytochrome P-450 Enzyme System