Van Wart H E
Department of Chemistry, Florida State University, Tallahassee 32306.
Matrix Suppl. 1992;1:31-6.
Human neutrophil collagenase (HNC) has been purified from extracts of fresh and outdated buffy coats and from the exudates of phorbol myristate acetate-stimulated neutrophils. The HNC present in the starting material from such preparations can be either latent or active, or have an approximate molecular weight of 75 or 58 kDa, depending upon whether the extraction buffer contains protease inhibitors and/or antioxidants. The purification of these different forms of HNC is described and is made possible by taking appropriate precautions to stabilize the HNC. For example, a purification protocol is described that allows the purification to homogeneity of the active and PCMB-active latent 58 kDa forms of HNC in high yield with specific collagenase activities that greatly exceed that of trypsin-activated human fibroblast collagenase (HFC). The pattern of activation of the latent 58 and 75 kDa species by trypsin, organomercurials and oxidants has been investigated. HNC is shown to preferentially hydrolyze type I over types II and III collagens in solution. The specificity of HNC toward the hydrolysis of 60 octapeptides has been examined and compared with HFC. HNC is shown to be a glycoprotein that contains complex N-linked oligosaccharides.
人中性粒细胞胶原酶（HNC）已从新鲜和过期的血沉棕黄层提取物以及佛波酯肉豆蔻酸酯刺激的中性粒细胞渗出物中纯化出来。根据提取缓冲液中是否含有蛋白酶抑制剂和/或抗氧化剂，此类制剂起始原料中存在的HNC可以是潜在的或活性的，或者分子量约为75或58 kDa。本文描述了这些不同形式HNC的纯化方法，并且通过采取适当措施稳定HNC使其成为可能。例如，描述了一种纯化方案，该方案能够以高产率将活性和PCMB活性的潜在58 kDa形式的HNC纯化至同质，其特定胶原酶活性大大超过胰蛋白酶激活的人成纤维细胞胶原酶（HFC）。研究了胰蛋白酶、有机汞化合物和氧化剂对潜在58 kDa和75 kDa形式的激活模式。结果表明，HNC在溶液中优先水解I型胶原蛋白而非II型和III型胶原蛋白。已检测HNC对60种八肽水解的特异性，并与HFC进行了比较。结果表明，HNC是一种含有复杂N-连接寡糖的糖蛋白。
