Smirnov A N, Shchelkunova T A
Biull Eksp Biol Med. 1989 Feb;107(2):179-82.
Estrophilic forms of rabbit liver cytosolic hydroxysteroid-dehydrogenase (HSD) were obtained as a highly purified preparations by means of fractionation with ammonium sulfate, gel-filtration, ion-exchange chromatography on DEAE-Sephadex A-50, affinity chromatography on estradiol-Sepharose and ion-exchange chromatography on DEAE-Toyopearl 650M. The protein express 4 different kinds of NADP-dependent activities: 3 alpha, 3 beta- and 17 beta-HSD activities with androgens and 20 alpha-HSD with progesterone as substrates. Revealed multiplicity of HSD enzymatic activity is demonstrated here for the first time. 17 beta-HSD activity of the protein preparations with estradiol is extremely low. Absence of a real metabolic activity of the protein with a ligand interacting with it rather intensively suggests that the isolated HSD forms can act not only as an enzyme, but also as a buffer-reserving mechanism for some steroids.
通过硫酸铵分级分离、凝胶过滤、在DEAE-葡聚糖A-50上进行离子交换色谱、在雌二醇-琼脂糖上进行亲和色谱以及在DEAE-托普雷斯650M上进行离子交换色谱，获得了兔肝细胞溶质羟类固醇脱氢酶（HSD）的嗜雌激素形式，并将其制备成高纯度制剂。该蛋白质表现出4种不同的依赖NADP的活性：以雄激素为底物时的3α、3β和17β-HSD活性，以及以孕酮为底物时的20α-HSD活性。蛋白质HSD酶活性的多重性在此首次得到证实。该蛋白质制剂与雌二醇的17β-HSD活性极低。该蛋白质与与其强烈相互作用的配体缺乏真正的代谢活性，这表明分离出的HSD形式不仅可以作为一种酶，还可以作为某些类固醇的缓冲储备机制。
