Purification of testosterone 5 alpha-reductase from human prostate by a four-step chromatographic procedure.
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.
Quemener
E
E
Département de Biochimie et Biologie Moléculaire, Faculté de Médecine, Brest, France.
Amet
Y
Y
di Stefano
S
S
Fournier
G
G
Floch
H H
HH
Abalain
J H
JH
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*
D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase