X-ray absorption studies of human matrix metalloproteinase-2 (MMP-2) bound to a highly selective mechanism-based inhibitor - Comparison with the latent and active forms of the enzyme
X-ray absorption studies of human matrix metalloproteinase-2 (MMP-2) bound to a highly selective mechanism-based inhibitor - Comparison with the latent and active forms of the enzyme
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DOI：
10.1074/jbc.m011604200
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发表时间：
2001-05-18
影响因子：
4.8
通讯作者：
Sagi, I
中科院分区：
文献类型：
--
作者：
Kleifeld, O;Kotra, LP;Sagi, I
恶性肿瘤表达高水平的锌依赖性内肽酶，称为基质金属蛋白酶（MMPs），被认为通过水解细胞外基质成分促进肿瘤转移和血管生成。在这些酶中，明胶酶A （MMP-2）和B （MMP-9）尤其与恶性过程有关，因此，它们已成为旨在阻断其活性的药物的靶标。因此，了解它们的分子结构是合理设计抑制剂的关键。在这里，我们对全长人MMP-2进行了潜伏、活性和抑制状态下的x射线吸收光谱分析，并报告了酶激活和抑制后锌离子位点的结构变化。我们还研究了MMP-2与SB-3CT复合物的分子结构，SB-3CT是最近报道的一种基于新机制的合成抑制剂，在明胶酶中具有高度选择性(1)。结果表明，SB-3CT直接与MMP-2的催化锌离子结合。有趣的是，抑制剂与催化锌结合的新模式将活性位点金属离子周围的构象环境重建回原酶的构象环境。
Malignant tumors express high levels of zinc-dependent endopeptidases called matrix metalloproteinases (MMPs), which are thought to facilitate tumor metastasis and angiogenesis by hydrolyzing components of the extracellular matrix. Of these enzymes, gelatinases A (MMP-2) and B (MMP-9), have especially been implicated in malignant processes, and thus, they have been a target for drugs designed to block their activity. Therefore, understanding their molecular structure is key for a rational approach to inhibitor design. Here, we have conducted x-ray absorption spectroscopy of the full-length human MMP-2 in its latent, active, and inhibited states and report the structural changes at the zinc ion site upon enzyme activation and inhibition. We have also examined the molecular structure of MMP-2 in com plex with SB-3CT, a recently reported novel mechanism-based synthetic inhibitor that was designed to be highly selective in gelatinases (1). It is shown that SB-3CT directly binds the catalytic zinc ion of MMP-2. Interestingly, the novel mode of binding of the inhibitor to the catalytic zinc reconstructs the conformational environment around the active site metal ion back to that of the proenzyme.