Harvima R J, Yabe K, Fräki J E, Fukuyama K, Epstein W L
J Invest Dermatol. 1987 Apr;88(4):393-7. doi: 10.1111/1523-1747.ep12469719.
Cathepsins B, D, H, and L were identified in the extract of 2-day-old rat epidermis and separated by gel filtration from aminoendopeptidase with a Mr of 400,000 and from the low-molecular-weight cysteine proteinase inhibitor. They were further purified by ion exchange column chromatography. The final separation for cathepsins B and H was performed by gel filtration, while cathepsin D was purified by pepstatin affinity chromatography and cathepsin L by fast protein liquid chromatography (FPLC). Substrate specificity, inhibitor susceptibility, and apparent molecular weights of the separated proteinases were determined and values compared to rat liver enzymes. Apparent molecular weights for epidermal cathepsins B, H, and L were higher than those for comparable liver enzymes of adult rats. The cysteine proteinase inhibitor in epidermis was found to inhibit cathepsins B, H, and L but not cathepsin D and aminoendopeptidase of rat epidermis. This study demonstrates the presence of cathepsin L in the epidermis and describes simultaneous separation and comparison of epidermal catheptic proteinases.
在2日龄大鼠表皮提取物中鉴定出组织蛋白酶B、D、H和L，并通过凝胶过滤将它们与分子量为400,000的氨基内肽酶以及低分子量半胱氨酸蛋白酶抑制剂分离。它们通过离子交换柱色谱进一步纯化。组织蛋白酶B和H的最终分离通过凝胶过滤进行，而组织蛋白酶D通过胃蛋白酶抑制剂亲和色谱纯化，组织蛋白酶L通过快速蛋白质液相色谱（FPLC）纯化。测定了分离出的蛋白酶的底物特异性、抑制剂敏感性和表观分子量，并将这些值与大鼠肝脏酶进行比较。表皮组织蛋白酶B、H和L的表观分子量高于成年大鼠相应肝脏酶的表观分子量。发现表皮中的半胱氨酸蛋白酶抑制剂可抑制组织蛋白酶B、H和L，但不抑制大鼠表皮的组织蛋白酶D和氨基内肽酶。本研究证明了表皮中存在组织蛋白酶L，并描述了表皮组织蛋白酶的同时分离和比较。
