Anderson R J, Babbitt L L, Liebentritt D K
Department of Medicine, Department of Veterans Affairs Medical Center, Omaha, Nebraska, USA.
Thyroid. 1995 Feb;5(1):61-6. doi: 10.1089/thy.1995.5.61.
To ascertain whether triiodothyronine (T3) sulfotransferase coeluted with the known phenol sulfotransferases (PSTs) during purification, human liver thermostable PST, thermolabile PST, and T3 sulfotransferase were assayed with p-nitrophenol, dopamine, and T3, respectively. Thermostable PST eluted from an ion-exchange column in two sequential peaks of activity (Peaks I and II), followed by a peak of thermolabile PST activity. There were three peaks of T3 sulfotransferase with thermostable PST: two within thermostable PST Peak I, and one peak of T3 sulfotransferase activity within thermostable PST Peak II. There was a minor peak of T3 sulfotransferase with thermolabile PST. Further purification of thermostable Peak I showed coelution of T3 sulfotransferase with thermostable PST during gel filtration and affinity chromatography. SDS-PAGE revealed a major protein band at 31 kDa. Dehydroepiandrosterone sulfotransferase comprised only 4% of the final activity. This report demonstrates coelution of T3 sulfotransferase with thermostable PST, shows a potential additional isozyme of T3 sulfotransferase, and points out the apparent minimal role of dehydroepiandrosterone sulfotransferase in T3 sulfation. The findings support the hypothesis that thermostable PST is the predominant human liver T3 sulfotransferase activity.
为确定在纯化过程中三碘甲状腺原氨酸（T3）磺基转移酶是否与已知的酚磺基转移酶（PSTs）一同洗脱，分别用人肝热稳定PST、热不稳定PST和T3磺基转移酶检测对硝基苯酚、多巴胺和T3。热稳定PST从离子交换柱上以两个连续的活性峰（峰I和峰II）洗脱，随后是热不稳定PST活性峰。T3磺基转移酶与热稳定PST有三个峰：热稳定PST峰I内有两个，热稳定PST峰II内有一个T3磺基转移酶活性峰。T3磺基转移酶与热不稳定PST有一个小峰。对热稳定峰I进一步纯化显示，在凝胶过滤和亲和层析过程中T3磺基转移酶与热稳定PST一同洗脱。SDS-PAGE显示在31 kDa处有一条主要蛋白带。脱氢表雄酮磺基转移酶仅占最终活性的4%。本报告证明T3磺基转移酶与热稳定PST一同洗脱，显示了T3磺基转移酶可能存在的另一种同工酶，并指出脱氢表雄酮磺基转移酶在T3硫酸化中明显起最小作用。这些发现支持热稳定PST是主要的人肝T3磺基转移酶活性这一假说。
