Zucker S, Turpeenniemi-Hujanen T, Ramamurthy N, Wieman J, Lysik R, Gorevic P, Liotta L A, Simon S R, Golub L M
Department of Medicine and Research, Veterans Administration Medical Center, Northport, NY 11768.
Biochem J. 1987 Jul 15;245(2):429-37. doi: 10.1042/bj2450429.
A metalloproteinase with activity against type IV collagen, type I collagen and gelatin has been purified from the cytosol of a highly metastatic mouse melanoma by anion-exchange, zinc-chelated and lectin-affinity column chromatography. The purified enzyme has a molecular mass of approx. 59 kDa and on isoelectric focusing in two-dimensional gels produced three spots with apparent isoelectric points (pI) between 5.7 and 6.1. Enzymic activity with collagen, but not gelatin, substrates was latent, requiring activation by trypsin or organomercurials. Trypsin activation of this metalloproteinase was accompanied by a change in molecular mass, whereas autoactivation after 1 month's storage, was not. The degradation of types I and IV collagen by the melanoma enzyme yielded products of lower molecular masses than those yielded by mammalian collagenases, this characteristic thus differentiating this metalloproteinase from classical collagenases.
一种对IV型胶原、I型胶原和明胶具有活性的金属蛋白酶已通过阴离子交换、锌螯合和凝集素亲和柱色谱法从高转移性小鼠黑色素瘤的胞质溶胶中纯化出来。纯化后的酶分子量约为59 kDa，在二维凝胶上进行等电聚焦时产生了三个表观等电点（pI）在5.7至6.1之间的斑点。该酶对胶原底物（而非明胶底物）的酶活性是潜伏性的，需要胰蛋白酶或有机汞化合物激活。这种金属蛋白酶经胰蛋白酶激活后分子量发生变化，而储存1个月后的自动激活则不会。黑色素瘤酶对I型和IV型胶原的降解产生的产物分子量低于哺乳动物胶原酶产生的产物，因此这一特性将这种金属蛋白酶与经典胶原酶区分开来。
