PURIFICATION AND PRELIMINARY CHARACTERIZATION OF STRATUM-CORNEUM CHYMOTRYPTIC ENZYME - A PROTEINASE THAT MAY BE INVOLVED IN DESQUAMATION
PURIFICATION AND PRELIMINARY CHARACTERIZATION OF STRATUM-CORNEUM CHYMOTRYPTIC ENZYME - A PROTEINASE THAT MAY BE INVOLVED IN DESQUAMATION
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DOI：
10.1111/1523-1747.ep12363804
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发表时间：
1993-08-01
影响因子：
6.5
通讯作者：
EGELRUD, T
中科院分区：
文献类型：
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作者：
EGELRUD, T
在最近的工作中，我们已经表明，丝氨酸蛋白酶，角质层胰凝乳蛋白酶，与在体外以及在体内的脱屑的作用相容的性质，通常存在于人角质层。角质层糜蛋白酶在KCl提取物的解离足底角质细胞的酶学性质进行了比较与其他已知的糜蛋白酶丝氨酸蛋白酶。角质层糜蛋白酶被发现显着不同牛糜蛋白酶，人组织蛋白酶G，和人肥大细胞糜蛋白酶方面的抑制剂和底物特异性。角质层糜蛋白酶进一步纯化分离的足底角质细胞的KCl提取物，通过凝胶上的亲和层析与共价连接的大豆胰蛋白酶抑制剂。纯化的制剂含有一种表观分子量为25 kD的主要组分和一种表观分子量略高的次要组分，如未还原的样品在聚丙烯酰胺凝胶中用十二烷基硫酸钠电泳后考马斯染色所示。这两种成分都与胰凝乳蛋白酶样活性有关，如在聚丙烯酰胺凝胶中与共聚酪蛋白的酶谱所示。在酶谱凝胶上，还发现纯化的制剂含有少量具有胰蛋白酶样活性的组分。主要纯化蛋白在还原和完全变性后具有约28 kD的表观分子量，并且显示含有碳水化合物。
In recent work we have shown that a serine proteinase, stratum corneum chymotryptic enzyme, with properties compatible with a role in desquamation in vitro as well as in vivo, is generally present in human stratum corneum. The enzymologic properties of the stratum corneum chymotryptic enzyme in a KCl extract of dissociated plantar corneocytes were compared with those of other known chymotryptic serine proteinases. Stratum corneum chymotryptic enzyme was found to differ significantly from bovine chymotrypsin, human cathepsin G, and human mast cell chymases in regard to inhibitor profile and substrate specificity. Stratum corneum chymotryptic enzyme was further purified from KCl extracts of dissociated plantar corneocytes by affinity chromatography on gels with covalently linked soybean trypsin inhibitor. The purified preparation contained one major component with apparent molecular weight 25 kD and one minor component with slightly higher apparent molecular weight as revealed by Coomassie staining after electrophoresis in polyacrylamide gels with sodium dodecyl sulphate of samples that had not been reduced. Both these components were associated with chymotrypsinlike activity as revealed by zymography in polyacrylamide gels with co-polymerized casein. On zymography gels, the purified preparation was also found to contain minor amounts of components with trypsinlike activity. The major purified protein had an apparent molecular weight of around 28 kD after reduction and full denaturation and was shown to contain carbohydrate.