Choi H Y, Hobkirk R
J Steroid Biochem. 1986 Dec;25(6):985-9. doi: 10.1016/0022-4731(86)90333-x.
Estrone sulfate sulfohydrolase (estrogen sulfatase) activity was solubilized by treatment with Triton X-100 from 105,000 g pellets of guinea pig uterus, testis and brain, as well as from rat liver and human placenta. The solubilized forms were subjected to chromatofocusing in the fast protein liquid chromatography (FPLC) system and on conventional columns packed in our laboratory. The guinea pig tissue pattern was complex. Uterus showed peaks of activity with apparent pI's of 9.11 and 7.6; testis contained 3 peaks with pI's of 9.18, 8.7 and 7.5; brain possessed peaks with pI's of 9.28 and 8.6. In each case the major activity peak was that with pI greater than 9. Rat liver activity chromatofocused as a single peak of apparent pI = 6.87 and the human placental enzyme also showed a single, though broad, peak, of pI = 6.57. This suggests not only that the guinea pig enzyme(s) differs markedly from those of rat liver and human placenta, but that there may be qualitative differences between the forms in the three guinea pig tissues. Chromatofocusing behaviour was not independent of the specific exchange resins and ampholytes utilized. The recovered enzyme activity was fairly stable and it seems that chromatofocusing could be a useful step in purification of the guinea pig enzyme(s), particularly the main form possessing a pI greater than 9.
用Triton X-100处理豚鼠子宫、睾丸和大脑的105,000 g沉淀，以及大鼠肝脏和人胎盘，可使硫酸雌酮硫酸酯水解酶（雌激素硫酸酯酶）活性溶解。将溶解后的形式在快速蛋白质液相色谱（FPLC）系统以及我们实验室装填的常规柱上进行色谱聚焦。豚鼠组织的图谱很复杂。子宫显示出活性峰，表观pI分别为9.11和7.6；睾丸含有3个峰，pI分别为9.18、8.7和7.5；大脑具有pI为9.28和8.6的峰。在每种情况下，主要活性峰的pI均大于9。大鼠肝脏活性经色谱聚焦后为一个表观pI = 6.87的单峰，人胎盘酶也显示出一个单一的（尽管较宽）峰，pI = 6.57。这不仅表明豚鼠的酶与大鼠肝脏和人胎盘的酶有明显差异，而且表明豚鼠三种组织中的酶形式可能存在质的差异。色谱聚焦行为并非独立于所使用的特定交换树脂和两性电解质。回收的酶活性相当稳定，似乎色谱聚焦可能是纯化豚鼠酶，特别是pI大于9的主要形式的有用步骤。
