TIMP/METALLOPROTEINASE STRUCTURE AND INTERACTIONS

描述：金属蛋白酶组织抑制物(TIMPs)抑制 基质金属蛋白酶对结缔组织的病理性水解作用 (MMPs)，它伴随着关节炎、癌症和退行性眼病。 关于TIMP如何与亚纳分子结合并使MMPs失活，目前知之甚少 亲和力。TIMP及其三维结构的可用性 与具有代表性的金属蛋白酶的络合物，在溶液中，将添加 理解高亲和力的结构基础。结构核磁共振 表征将有助于TIMP衍生抑制剂的合理开发 开发更好、更具选择性的MMPs抑制剂的长期努力。 具体目的是：(1)细化抑制剂的核磁共振结构 人TIMP-1(N-TIMP-1)的结构域到高分辨率。(2)绘制曲面地图 被人基质分解蛋白-1催化结构域(MMP3(DC))干扰的N-TIMP-1， 使用核磁共振技术来创造一个“足迹”。(3)测量和解读密切接触 在N-TIMP-1和基质金属蛋白酶-3(DC)之间需要定位它们的结合 界面。(4)识别并纠正发生的构象变化 在基质金属蛋白酶-3(DC)和N-TIMP-1络合的溶液结构中。 (5)对接修订的N-TIMP-1和基质金属蛋白酶-3(DC)溶液结构。(6) N-TIMP-1与基质金属蛋白酶-3(DC)结合的DCP与埋藏的比较 溶液结构中发现的界面表面积和极性。(7) 比较基质金属蛋白酶-3(DC)存在和不存在时N-TIMP-1骨架的动力学变化。 (8)探索N-TIMP-1 Thr2到Leu取代的途径 引入基质金属蛋白酶-3(DC)和基质金属蛋白酶-1(DC)(胶原酶)之间的选择性 催化区域)，使用滴定量热法和核磁共振。

DESCRIPTION: Tissue inhibitors of metalloproteinases (TIMPs) inhibit pathological hydrolysis of connective tissue by matrix metalloproteinases (MMPs) which accompanies arthritis, cancer, and degenerative eye diseases. Little is known of how TIMPs bind and inactivate MMPs with subnanomolar affinity. Availability of three-dimensional structures of a TIMP and of its complex with a representative metalloproteinase, in solution, would add understanding of the structural basis of high affinity. Structural NMR characterization will aid rational development of TIMP-derived inhibitors in long-range efforts to develop better, more selective inhibitors of MMPs. The specific aims are: (1) Refine the NMR structure of the inhibitory domain of human TIMP-1 (N-TIMP-1) to high resolution. (2) Map the surface of N-TIMP-1 perturbed by human stromelysin-1 catalytic domain (MMP-3(DC)), using NMR to create a "footprint". (3) Measure and interpret close contacts between N-TIMP-1 and MMP-3(DC) needed to orient them about their binding interface. (4) Identify and correct for conformational changes which occur in the solution structures of MMP-3(DC) and of N-TIMP-1 upon complexation. (5) Dock the revised N-TIMP-1 and MMP-3(DC) solution structures. (6) Compare DCp of the association of N-TIMP-1 with MMP-3(DC) with the buried interfacial surface area and polarity found in the solution structure. (7) Compare N-TIMP-1 backbone dynamics in the absence and presence of MMP-3(DC). (8) Probe the means by which the Thr2-to-Leu substitution of N-TIMP-1 introduces selectivity between MMP-3(DC) and MMP-1(DC) (collagenase catalytic domain), using titration calorimetry and NMR.