Matrix Metalloproteinase Inhibition and Specificity

基质金属蛋白酶抑制和特异性

• 批准号: 7118809
• 负责人: Steven R Van Doren
• 金额: $ 30.35万
• 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 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7118809
• 关键词: active sites; biological signal transduction; calorimetry; chemical binding; collagenase; conformation; elastases; elastin; enzyme activity; enzyme inhibitors; enzyme substrate complex; flavonoids; metalloendopeptidases; molecular site; nuclear magnetic resonance spectroscopy; protein structure function; stromelysin; thermodynamics; tissue inhibitor of metalloproteinases

DESCRIPTION (provided by applicant): Matrix metalloproteinases (MMPs), and TIMPs that inhibit them, control tissue remodeling in cancer, arthritis, arterial disease, pulmonary disease, reproductive events, and wound healing. Mechanisms of action of two key natural MMP inhibitors are not adequately understood, i.e., the human protein TIMP-1 and the cancer preventative EGCG from green tea. The mechanisms of action of these two with therapeutic potential will be investigated further. The previous project found the high affinity of a typical TIMP and MMP to be driven by entropy gain, largely configurational. The backbone of the beta-barrel of N-TIMP-1 is mobilized upon binding of MMP-3. For a more complete spatial and temporal view of flexibility changes linked to their binding, NMR studies of mobility will be expanded to free and N-TIMP-1-bound states of MMP-3, to side chain methyl groups, and to longer time scales where thermodynamic inferences become more robust. Engineering of N-TIMP-1 to enhance therapeutic potential has neglected its undesirable growth factor activity. The previous project found a conserved surface on TIMPs hypothesized to mediate this activity. This patch will be mutagenized in an effort to remove N-TIMP-1's activity of stimulating cell proliferation. Signaling pathways and receptor mediating TIMP-1's growth factor activity will be identified. Inhibition of a few MMPs was found among the chemopreventative bioactivities of the main polyphenol in green tea, EGCG. Which MMPs are inhibited by EGCG will be surveyed. The novel mechanism of MMP inhibition by EGCG and its structural mode of binding an MMP will be investigated. A contemporary drug target for cardiovascular and pulmonary conditions is MMP-12. MMP-12 has distinctively high activity upon the elastin component of blood vessels and lungs and upon the elastin-preserving inhibitor of elastase known as a1-anti-trypsin. Sequence determinants of MMP-12's activity upon elastin and a1-anti-trypsin are not known and will be probed. Sites in MMP-12 that interact with a1-anti-trypsin will be mapped by NMR. Elastase specificity will be engineered out of MMP-12, and then transferred into MMP-3. Discovery of features that confer the unique specificity to MMP-12 should aid design of more selective inhibitors and may suggest determinants of specificities of other MMPs.

描述(申请人提供)：基质金属蛋白酶(MMPs)和抑制它们的TIMP，控制癌症、关节炎、动脉疾病、肺部疾病、生殖事件和伤口愈合中的组织重塑。两种关键的天然基质金属蛋白酶抑制剂的作用机制尚不清楚，即人类蛋白TIMP-1和绿茶中的防癌EGCG。这两种具有治疗潜力的药物的作用机制有待进一步研究。上一个项目发现，典型的TIMP和MMP的高亲和力是由熵增益驱动的，很大程度上是由构型决定的。当与基质金属蛋白酶-3结合时，N-TIMP-1的β-桶的骨架被动员。为了获得与其结合相关的灵活性变化的更完整的空间和时间视图，对迁移率的核磁共振研究将扩展到基质金属蛋白酶-3的自由和N-TIMP-1结合状态，侧链甲基，以及更长的时间尺度，在那里热力学推断变得更加可靠。为提高治疗潜力而设计的N-TIMP-1忽略了其不良的生长因子活性。先前的项目在TIMP上发现了一个保守的表面，假设介导了这一活动。该贴片将被诱变，以努力去除N-TIMP-1‘S刺激细胞增殖的活性。介导TIMP-1‘S生长因子活性的信号通路和受体将被确定。在绿茶中主要的多酚类化合物EGCG的化学预防生物活性中，发现了少数MMPs的抑制作用。哪些MMPs被EGCG抑制，将被调查。我们将探讨EGCG抑制基质金属蛋白酶的新机制及其与基质金属基质结合的结构模式。目前治疗心血管和肺部疾病的药物靶点是基质金属蛋白酶-12。基质金属蛋白酶-12对血管和肺的弹性蛋白成分以及弹性蛋白保留的弹性蛋白酶抑制剂A1-抗胰蛋白酶具有明显的高活性。基质金属蛋白酶-12‘S活性对弹性蛋白和A1-抗胰蛋白酶活性的序列决定因素尚不清楚，将进行探讨。在基质金属蛋白酶-12中与A1-抗胰蛋白酶相互作用的位置将通过核磁共振绘制出来。弹性蛋白酶的特异性将从基质金属蛋白酶-12中分离出来，然后转移到基质金属蛋白酶-3中。发现赋予基质金属蛋白酶-12独特特异性的特征应该有助于设计更具选择性的抑制剂，并可能提出其他基质金属蛋白酶特异性的决定因素。