Schechter N M, Fräki J E, Geesin J C, Lazarus G S
J Biol Chem. 1983 Mar 10;258(5):2973-8.
A chymotrypsin-like proteinase was purified 2400-fold from human skin. The procedure involves extraction of the proteinase from skin in 2 M KCl, precipitation with protamine chloride, fractionation by gel filtration chromatography, and fractionation by chromatography using a CH-Sepharose-D-tryptophan methyl ester affinity column. The properties of this proteinase were compared to the rat mast cell proteinase I and human cathepsin G. Differences were observed in the rates at which the proteinases were inhibited by diisopropyl fluorophosphate, the sensitivity of the proteinases to protein proteolytic inhibitors, the relative hydrolytic rates of the proteinases for a series of substrates, and the kinetic constants of the proteinases for synthetic substrates. The human skin proteinase did not react with antiserum to the rat skin proteinase and did not elute in the same position as the rat skin proteinase on gel filtration columns. These data demonstrate that the human skin proteinase is distinct from the other proteinases. Extracts of involved skin from patients with cutaneous mastocytosis had 15-fold higher levels of chymotryptic activity than extracts of uninvolved skin or skin from normal controls. The enzymatic properties of the material extracted from the biopsied skin were similar to those of the proteinase from normal skin, suggesting that the human skin chymotrypsin-like proteinase is a mast cell constituent.
从人皮肤中纯化出一种胰凝乳蛋白酶样蛋白酶，纯化倍数达2400倍。该纯化过程包括在2M氯化钾中从皮肤提取蛋白酶、用鱼精蛋白氯化物沉淀、通过凝胶过滤色谱法分级分离以及使用CH-琼脂糖-D-色氨酸甲酯亲和柱进行色谱法分级分离。将这种蛋白酶的特性与大鼠肥大细胞蛋白酶I和人组织蛋白酶G进行了比较。观察到在二异丙基氟磷酸抑制蛋白酶的速率、蛋白酶对蛋白质水解抑制剂的敏感性、蛋白酶对一系列底物的相对水解速率以及蛋白酶对合成底物的动力学常数方面存在差异。人皮肤蛋白酶不与大鼠皮肤蛋白酶的抗血清发生反应，并且在凝胶过滤柱上的洗脱位置与大鼠皮肤蛋白酶不同。这些数据表明人皮肤蛋白酶与其他蛋白酶不同。皮肤肥大细胞增多症患者受累皮肤的提取物中胰凝乳蛋白酶活性水平比未受累皮肤或正常对照皮肤的提取物高15倍。从活检皮肤中提取的物质的酶学特性与正常皮肤蛋白酶的特性相似，这表明人皮肤胰凝乳蛋白酶样蛋白酶是肥大细胞的一种成分。
