Hobkirk R, Girard L R, Durham N J, Khalil M W
Biochim Biophys Acta. 1985 Apr 5;828(2):123-9. doi: 10.1016/0167-4838(85)90047-0.
The estrogen sulfotransferase activity of high-speed supernatants of mouse placenta and uterus behaves on conventional and high-performance liquid chromatographic gel filtration as an enzyme species with a molecular weight of the order of 30 000. This is so whether the cytosols are freshly prepared or have been stored at -20 degrees C before chromatography. The presence of thiol groups or EDTA has no effect on the elution pattern. The partially purified enzyme is extremely unstable and is poorly recovered by (NH4)2SO4 fractionation. Some stabilization can be achieved in the presence of 0.1 microM estradiol. Chromatofocusing of cytosols results in the elution of one or two sulfotransferase peaks, depending upon experimental conditions such as the presence or absence of thiol groups. These peaks act upon both estrone and estradiol as substrates. Chromatofocusing by fast protein liquid chromatography (FPLC) in the absence of thiol groups results in the elution of one sulfotransferase peak whose activity can be detected only when thiol groups are present during enzyme assay.
小鼠胎盘和子宫高速上清液的雌激素磺基转移酶活性，在常规和高效液相色谱凝胶过滤中表现为一种分子量约为30000的酶种类。无论胞质溶胶是新鲜制备的，还是在色谱分析前已保存在-20℃，情况都是如此。巯基或EDTA的存在对洗脱模式没有影响。部分纯化的酶极其不稳定，通过硫酸铵分级分离回收率很低。在0.1微摩尔雌二醇存在的情况下可以实现一定程度的稳定。胞质溶胶的色谱聚焦会导致一个或两个磺基转移酶峰的洗脱，这取决于实验条件，如巯基的存在与否。这些峰以雌酮和雌二醇作为底物起作用。在没有巯基的情况下通过快速蛋白质液相色谱（FPLC）进行色谱聚焦，会导致一个磺基转移酶峰的洗脱，其活性只有在酶测定过程中存在巯基时才能被检测到。
