Neshkova E A, Dotsenko V L, Larionova N I, Iarovaia G A
Biokhimiia. 1993 Dec;58(12):1886-92.
A procedure for one-step rapid isolation of highly purified elastase and cathepsin G from human leucocytes including biospecific chromatography on Gordox-Sepharose is described. Electrophoresis in gradient (10-15%) polyacrylamide gel in the presence of sodium dodecyl sulfate revealed three elastase isoforms with molecular masses of 26.3, 27.8 and 28.9 kDa. Cathepsin G produced one protein band with a 27 kDa mobility. The pH optimum for elastase and cathepsin G are 7.0-7.5 and 7.5-8.0, respectively. The specific activities of elastase and cathepsin G preparations as measured by the p-nitrophenyl ester-tert butoxycarbonyl-L-alanine and the ethyl ester-benzoyl tyrosine, hydrolysis are equal to 3-8 and 20-40 mumol/min/mg protein, respectively.
本文描述了一种从人白细胞中一步快速分离高纯度弹性蛋白酶和组织蛋白酶G的方法，包括在Gordox-Sepharose上进行生物特异性色谱分离。在十二烷基硫酸钠存在下，在梯度（10-15%）聚丙烯酰胺凝胶中进行电泳，结果显示三种弹性蛋白酶同工型，其分子量分别为26.3、27.8和28.9 kDa。组织蛋白酶G产生一条迁移率为27 kDa的蛋白带。弹性蛋白酶和组织蛋白酶G的最适pH分别为7.0-7.5和7.5-8.0。通过对硝基苯酯-叔丁氧羰基-L-丙氨酸和乙酯-苯甲酰酪氨酸的水解来测定，弹性蛋白酶和组织蛋白酶G制剂的比活性分别为3-8和20-40 μmol/min/mg蛋白质。
