TIMP/METALLOPROTEINASE STRUCTURE AND INTERACTIONS

TIMP/金属蛋白酶结构和相互作用

• 批准号: 6180511
• 负责人: Steven R Van Doren
• 金额: $ 21.87万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2002-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6180511
• 关键词: active sites; analytical method; calorimetry; chemical binding; collagenase; conformation; enzyme substrate complex; molecular site; nuclear magnetic resonance spectroscopy; protein structure function; solutions; stromelysin; thermodynamics; tissue inhibitor of metalloproteinases

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.

描述：金属蛋白酶（TIMP）的组织抑制剂抑制 基质金属蛋白酶对结缔组织的病理水解 （MMP）伴随关节炎，癌症和退化性眼部疾病。 鲜为人知的TIMP如何结合和灭活MMP 亲和力。 TIMP及其三维结构的可用性 在溶液中，具有代表性金属蛋白酶的复合物将增加 了解高亲和力的结构基础。 结构NMR 表征将有助于合理发展TIMP衍生的抑制剂 远程努力为MMP的更好，更有选择性的抑制剂发展。 具体目的是：（1）完善抑制的NMR结构 人类TIMP-1（N-TIMP-1）的领域至高分辨率。 （2）绘制表面 n-Timp-1受人类stromelysin-1催化结构域（MMP-3（DC））的干扰 使用NMR创建“足迹”。 （3）测量和解释密切联系 在N-TIMP-1和MMP-3（DC）之间，需要将它们定向到它们的结合 界面。 （4）识别并纠正发生的构象变化 在络合后，MMP-3（DC）和N-TIMP-1的溶液结构中。 （5）对接修订后的N-TIMP-1和MMP-3（DC）溶液结构。 （6） 比较N-TIMP-1与MMP-3（DC）与埋藏的DCP 在溶液结构中发现的界面表面积和极性。 （7） 在不存在和存在MMP-3（DC）的情况下比较N-TIMP-1骨干动力学。 （8）探测n-Timp-1 th2-th-leu替换的手段 引入MMP-3（DC）和MMP-1（DC）（胶原酶）之间的选择性 催化域），使用滴定量热法和NMR。