The role of the c-terminal domain of human collagenase-3 (MMP-13) in the activation of procollagenase-3, substrate specificity, and tissue inhibitor of metalloproteinase interaction
The role of the c-terminal domain of human collagenase-3 (MMP-13) in the activation of procollagenase-3, substrate specificity, and tissue inhibitor of metalloproteinase interaction
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DOI：
10.1074/jbc.272.12.7608
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发表时间：
1997-03-21
影响因子：
4.8
通讯作者：
Murphy, G
中科院分区：
文献类型：
--
作者：
Knauper, V;Cowell, S;Murphy, G
重组人胶原原酶-3及其C端截短型（Delta（249-451）procollagenase-3）在骨髓瘤细胞中得到了稳定表达并纯化，该截短的酶原可被乙酸氨基苯汞以一种短寿命的中间体形式加工（N-末端Leu（58））转化为最终活性形式（N-末端Tyr（85）），活化动力学不受血红素样C-末端结构域的去除的影响，使用两种淬灭的荧光底物，发现胶原酶-3和Delta（249-451）胶原酶-3的比活性相似，但Delta（249-451）胶原酶-3不能切割天然三螺旋胶原然而，注意到I型胶原的β 1，2（I）链对Delta（249-451）胶原酶-3敏感，这表明催化结构域显示端肽酶活性，从而产生比天然I型胶原蛋白中的α 1，2（I）链稍短的α 1，2（I）链。可以得出结论，C-末端结构域仅是胶原酶-3的三重解旋酶活性所必需的，前胶原酶-3和活性胶原酶-3与I型胶原蛋白的结合是由C-末端结构域介导的，胶原酶-3和Delta（249-451）胶原酶-3都水解大的腱生蛋白C同种型、纤连蛋白、重组纤连蛋白片段和IV、IX、X和XIV型胶原;因此，这些事件与C-末端结构域相互作用无关。相反，较小的软骨XI型胶原对切割具有抗性，对野生型和
Recombinant human procollagenase-3 and a C-terminal truncated form (Delta(249-451) procollagenase-3) have been stably expressed in myeloma cells and purified, The truncated proenzyme could be processed by amino-phenylmercuric acetate via a short lived intermediate form (N-terminal Leu(58)) to the final active form (N-terminal Tyr(85)), The kinetics of activation were not affected by removal of the hemopexin-like C-terminal domain, The specific activities of both collagenase-3 and Delta(249-451) collagenase-3 were found to be similar using two quenched fluorescent substrates, but Delta(249-451) collagenase-3 failed to cleave native triple helical collagens (types I and II) into characteristic one- and three-quarter fragments, It was noted, however, that the beta 1,2(I) chains of type I collagen were susceptible to Delta(249-451) collagenase-3, which indicates that the catalytic domain displays telopeptidase activity, thereby generating alpha 1,2(I) chains that are slightly shorter than those in native type I collagen, It can be concluded that the C-terminal domain is only essential for the triple helicase activity of collagenase-3, Binding of procollagenase-3 and active collagenase-3 to type I collagen is mediated by the C-terminal domain, Both collagenase-3 and Delta(249-451) collagenase-3 hydrolyzed the large tenascin C isoform, fibronectin, recombinant fibronectin fragments, and type IV, IX, X, and XIV collagens; thus, these events were independent from C-terminal domain inter actions, In contrast, the minor cartilage type XI collagen was resistant to cleavage, Kinetic analysis of the mechanism of inhibition of wild-type and