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（由申请人提供）：基质金属蛋白酶（MMP）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的组织重塑。两种关键天然MMP抑制剂的作用机制尚未充分理解，即人蛋白TIMP-1和绿茶的癌症预防EGCG。这两个具有治疗潜力的作用机制将进一步研究。以前的项目发现，典型的TIMP和MMP的高亲和力是由熵增益驱动的，这在很大程度上是配置。 N-TIMP-1的Beta桶的骨架在MMP-3结合后动员。为了使其具有与其结合有关的灵活性变化的更完整的空间和时间视图，NMR的迁移率研究将扩展到MMP-3的自由和N-TIMP-1结合状态，侧链甲基组以及更长的时间尺度，其中热力学推断变得更加可靠。 N-TIMP-1的工程以增强治疗潜力已忽略了其不良的生长因子活性。先前的项目发现，在假设介导该活动的TIMP上保守了表面。该斑块将被诱变，以消除N-TIMP-1刺激细胞增殖的活性。将确定信号通路和受体介导的TIMP-1生长因子活性。在绿茶中，EGCG的主要多酚的化学预防生物活性率中发现了一些MMP。 EGCG抑制哪些MMP。 EGCG抑制MMP的新机制及其结合MMP的结构模式。心血管和肺部疾病的当代药物靶标是MMP-12。 MMP-12对血管和肺的弹性蛋白成分以及弹性蛋白抑制剂的弹性蛋白具有明显的活性，称为A1-抗tryprypsin。 MMP-12在弹性蛋白和a1-抗trypsin上活性的序列决定因素尚不清楚，并且将被探测。 NMR将映射与A1-Anti-Trypsin相互作用的MMP-12中的位点。弹性酶特异性将从MMP-12中设计，然后转移到MMP-3中。发现赋予MMP-12独特特异性的特征应有助于设计更多选择性抑制剂，并可能表明决定其他MMP的特异性。