Lööf L, Hjertén S
Biochim Biophys Acta. 1980 Feb 22;617(2):192-204. doi: 10.1016/0005-2760(80)90162-9.
An enzyme that catalyzes the transfer of sulphate from 3'-phosphoadenosine 5'-phosphosulphate to bile salts was purified from human liver cytosol by chromatography on DEAE-Sephadex and Sephadex G-200, by agarose suspension electrophoresis and by isoelectric focusing in free solution. The purified enzyme was also active towards oestrone, dehydroepiandrosterone and phenol. No other liver steroid sulphotransferases could be detected during this purification procedure. Km values of 1.8 . 10(-6) M and 3.3 . 10(-6) M for glycolithocholate and 3'-phosphoadenosine 5'-phosphosulphate respectively were found. The sulphotransferase has an isoelectric point of 5.5. The enzyme was markedly activated by Mg2+, Mn2+ and Co2+ and inhibited by Cu2+, Fe2+ and Zn2+. Chenodeoxycholate and deoxycholate were sulphated at the 7-OH and 12-OH position, respectively. No bile salt disulphate formation was detected. A 30-fold increase in specific activity was obtained, although the purification based on ultraviolet light measurements was considerably higher.
通过在DEAE-葡聚糖凝胶和葡聚糖凝胶G-200上进行色谱分析、琼脂糖悬浮电泳以及在自由溶液中进行等电聚焦，从人肝细胞溶胶中纯化出一种催化硫酸根从3'-磷酸腺苷5'-磷酸硫酸转移至胆汁盐的酶。纯化后的酶对雌酮、脱氢表雄酮和苯酚也有活性。在此纯化过程中未检测到其他肝脏类固醇硫酸转移酶。分别测得甘氨石胆酸和3'-磷酸腺苷5'-磷酸硫酸的Km值为1.8×10⁻⁶M和3.3×10⁻⁶M。该硫酸转移酶的等电点为5.5。该酶被Mg²⁺、Mn²⁺和Co²⁺显著激活，被Cu²⁺、Fe²⁺和Zn²⁺抑制。鹅去氧胆酸和脱氧胆酸分别在7-OH和12-OH位被硫酸化。未检测到胆汁盐二硫酸盐的形成。尽管基于紫外光测量的纯化程度要高得多，但比活性提高了30倍。
