Kirschke H, Schmidt I, Wiederanders B
Biochem J. 1986 Dec 1;240(2):455-9. doi: 10.1042/bj2400455.
Cathepsin S was purified from bovine spleen by acid autolysis, (NH4)2SO4 fractionation and chromatography on CM-Sephadex C-50, CM-cellulose and activated-thiol-Sepharose. Cathepsin L was isolated from lysosomal fractions of rat liver, rat kidney and bovine liver. Generally, cathepsin L was bound tightly to CM-Sephadex C-50. Preparations of cathepsin L from rat liver, rat kidney and bovine liver were shown to have kinetic constants for the substrate benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide in the same range (Km 2-3 microM). Benzyloxycarbonyl-Phe-Phe-diazomethane proved to be a sensitive irreversible inhibitor of cathepsin L from different species. Cathepsin S differed in all these characteristics from cathepsin L. A polyclonal antibody to cathepsin L from rat reacted with bovine cathepsin L but not with bovine cathepsin S.
组织蛋白酶S通过酸自溶、硫酸铵分级分离以及在CM-葡聚糖凝胶C-50、CM-纤维素和活化硫醇-琼脂糖凝胶上进行层析从牛脾脏中纯化得到。组织蛋白酶L从大鼠肝脏、大鼠肾脏和牛肝脏的溶酶体组分中分离出来。一般来说，组织蛋白酶L与CM-葡聚糖凝胶C-50紧密结合。从大鼠肝脏、大鼠肾脏和牛肝脏中制备的组织蛋白酶L对底物苄氧羰基-苯丙氨酸-精氨酸-7-(4-甲基)香豆素酰胺的动力学常数在相同范围内（Km为2 - 3微摩尔）。苄氧羰基-苯丙氨酸-苯重氮甲烷被证明是不同物种组织蛋白酶L的一种敏感的不可逆抑制剂。组织蛋白酶S在所有这些特性上与组织蛋白酶L不同。来自大鼠的针对组织蛋白酶L的多克隆抗体与牛组织蛋白酶L反应，但不与牛组织蛋白酶S反应。
