Falany C N, Vazquez M E, Kalb J M
Department of Pharmacology and Cancer Center, University of Rochester, NY 14642.
Biochem J. 1989 Jun 15;260(3):641-6. doi: 10.1042/bj2600641.
A form of sulphotransferase capable of sulphating dehydroepiandrosterone and other steroids was purified from cytosol prepared from human liver. Dehydroepiandrosterone sulphotransferase was purified 621-fold when compared with the activity in cytosol using DEAE-Sepharose CL-6B and adenosine 3',5'-bisphosphate-agarose affinity chromatography. During affinity chromatography, dehydroepiandrosterone sulphation activity could be resolved from p-nitrophenol sulphation activity catalysed by phenol sulphotransferase by using a gradient of adenosine 3'-phosphate 5'-phosphosulphate. The purified enzyme was most active towards dehydroepiandrosterone but was capable of conjugating a number of other steroids, including pregnenolone, androsterone and beta-oestradiol. No activity towards p-nitrophenol or dopamine, substrates for the phenol sulphotransferase, was observed with the pure enzyme. A single band with a subunit molecular mass of 35 kDa was observed by Coomassie Blue staining following SDS/polyacrylamide-gel electrophoresis of the purified enzyme. A molecular mass of 68-70 kDa was calculated for the active form of the enzyme by chromatography on Sephacryl S-200, suggesting that the active form of the enzyme is a dimer.
一种能够将脱氢表雄酮和其他类固醇硫酸化的磺基转移酶，是从人肝脏制备的胞质溶胶中纯化得到的。与胞质溶胶中的活性相比，使用DEAE-琼脂糖CL-6B和腺苷3',5'-二磷酸琼脂糖亲和色谱法，脱氢表雄酮磺基转移酶被纯化了621倍。在亲和色谱过程中，通过使用腺苷3'-磷酸5'-磷酸硫酸酯梯度，可以将脱氢表雄酮硫酸化活性与由酚磺基转移酶催化的对硝基苯酚硫酸化活性分离。纯化后的酶对脱氢表雄酮活性最高，但也能够结合许多其他类固醇，包括孕烯醇酮、雄酮和β-雌二醇。用纯酶未观察到对酚磺基转移酶的底物对硝基苯酚或多巴胺的活性。对纯化后的酶进行SDS/聚丙烯酰胺凝胶电泳后，考马斯亮蓝染色观察到一条亚基分子量为35 kDa的条带。通过在Sephacryl S-200上进行色谱分析，计算出该酶活性形式的分子量为68-70 kDa，表明该酶的活性形式是二聚体。
