Stevens R L, Fluharty A L, Skokut M H, Kihara H
J Biol Chem. 1975 Apr 10;250(7):2495-501.
Arylsulfatase A (cerebroside sulfate sulfohydrolase) was purified 3500-fold at a 7% yield from human urine. A crude urinary protein concentrate was prepared by treating pooled urine with ammonium sulfate and subsequently drying the precipitate with acetone. The powder thus obtained was extracted with buffer and was subjected to chromatographic and electrophoretic procedures as follows: (a) ammonium sulfate reverse gradient solubilization chromatography; (b) DEAE-cellulose chromatography; (c) Sephadex G-200 gel filtration; (d) preparative polyacrylamide gel electrophoresis; (e) SP-Sephadex chromatography; and (f) antialbumin-Sepharose chromatography. The enzyme was judged to be essentially homogeneous by: (a) a single band on polyacrylamide gel electrophoresis at two pH values; (b) formation of a single precipitin line on immunodiffusion against its antiserum: (c) complete freedom from albumin, the major contaminating protein; and (d) a single band on sodium dodecyl sulfate gel electrophoresis.
芳基硫酸酯酶A（脑苷脂硫酸酯硫酸水解酶）从人尿中以7%的产率纯化了3500倍。通过用硫酸铵处理混合尿液，随后用丙酮干燥沉淀物，制备粗制尿蛋白浓缩物。将由此获得的粉末用缓冲液提取，并进行如下色谱和电泳程序：（a）硫酸铵反向梯度溶解色谱法；（b）DEAE - 纤维素色谱法；（c）Sephadex G - 200凝胶过滤法；（d）制备性聚丙烯酰胺凝胶电泳；（e）SP - Sephadex色谱法；以及（f）抗白蛋白 - 琼脂糖色谱法。通过以下方法判断该酶基本纯一：（a）在两个pH值下聚丙烯酰胺凝胶电泳上出现单一条带；（b）在针对其抗血清的免疫扩散中形成单一沉淀线；（c）完全不含主要污染蛋白白蛋白；以及（d）十二烷基硫酸钠凝胶电泳上出现单一条带。
