Quemener E, Amet Y, di Stefano S, Fournier G, Floch H H, Abalain J H
Département de Biochimie et Biologie Moléculaire, Faculté de Médecine, Brest, France.
Steroids. 1994 Dec;59(12):712-8. doi: 10.1016/0039-128x(94)90103-1.
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.
通过0.5%正辛基-β-D-吡喃葡萄糖苷从人前列腺组织中以活性形式溶解的核膜结合睾酮5α-还原酶，经包括DEAE-三乙醇胺离子交换、羟基磷灰石吸附、睾酮-琼脂糖亲和及琼脂糖4B凝胶过滤的四步色谱程序进行纯化。根据比酶活性的增加判断，实现了约30倍的纯化。我们纯化了酸性pH最适的2型5α-还原酶同工酶。通过SDS-PAGE估计纯化酶的表观分子量为42,000。同时，我们分离出一种38 kDa的蛋白质，其特征在于对睾酮具有真正的亲和力，并可能与5α-还原酶相关联。
