Masure S, Billiau A, Van Damme J, Opdenakker G
Rega Institute for Medical Research, University of Leuven, Belgium.
Biochim Biophys Acta. 1990 Sep 24;1054(3):317-25. doi: 10.1016/0167-4889(90)90103-k.
Several human cell lines were studied for the production of gelatinases. Diploid fibroblasts, the melanoma cell line Bowes, the MG-63 osteosarcoma cell line and the human hepatoma cell line Malavu all constitutively produced a 67 kDa gelatinase. Gelatinolytic enzymes were quantified by a sensitive zymographic substrate conversion assay. Upon induction with phorbol 12-myristate 13-acetate (PMA), the human hepatoma cell line secreted considerable amounts of an 85 kDa gelatinase activity. The induction process was time- and dose-dependent. It represented a true increase in production per individual cell and was associated by a marked change of the cell morphology. The effect of various proteinase inhibitors and the maximal activity of the enzyme near neutral pH demonstrate that it is a neutral metalloproteinase. Characterization studies showed the 85 kDa gelatinase to be transformed to lower molecular weight, active forms by treatment with p-aminophenylmercuric acetate (APMA) or trypsin.
对几种人类细胞系进行了明胶酶产生情况的研究。二倍体成纤维细胞、黑色素瘤细胞系Bowes、MG - 63骨肉瘤细胞系和人类肝癌细胞系Malavu均组成性地产生一种67 kDa的明胶酶。通过灵敏的酶谱底物转化分析对明胶分解酶进行定量。用佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）诱导后，人类肝癌细胞系分泌大量85 kDa的明胶酶活性。诱导过程具有时间和剂量依赖性。它代表单个细胞产量的真正增加，并且与细胞形态的显著变化相关。各种蛋白酶抑制剂的作用以及该酶在接近中性pH时的最大活性表明它是一种中性金属蛋白酶。特性研究表明，用对氨基苯基汞乙酸盐（APMA）或胰蛋白酶处理后，85 kDa的明胶酶可转化为较低分子量的活性形式。
