title: Purification of testosterone 5 alpha-reductase from human prostate by a four-step chromatographic procedure.
abstract: Nuclear membrane bound testosterone 5 alpha-reductase solubilized in active form from human prostatic tissue by 0.5% n-octyl beta-D-glucopyranoside was purified by a four-step chromatographic procedure including DEAE-Trisacryl ion exchange, hydroxylapatite adsorption, testosterone-Sepharose affinity and Sepharose 4B gel filtration. A purification of approximately 30-fold was achieved judging from the increase in the specific enzymatic activity. We have purified the acidic pH-optimum 5 alpha-reductase type 2 isoenzyme. The apparent molecular weight of the purified enzyme was estimated as 42,000 by SDS-PAGE. At the same time we isolated a 38 kDa protein characterized by a real affinity for testosterone and by a possible association to the 5 alpha-reductase enzyme.
authors.affiliation: Département de Biochimie et Biologie Moléculaire, Faculté de Médecine, Brest, France.
authors.forename: E
authors.initials: E
authors.lastname: Quemener
authors.forename: Y
authors.initials: Y
authors.lastname: Amet
authors.forename: S
authors.initials: S
authors.lastname: di Stefano
authors.forename: G
authors.initials: G
authors.lastname: Fournier
authors.forename: H H
authors.initials: HH
authors.lastname: Floch
authors.forename: J H
authors.initials: JH
authors.lastname: Abalain
mesh_terms: D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D008970:Molecular Weight; D011467:Prostate / Q000201:enzymology*
chemical_list: D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase