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中的组织重塑。两种关键的天然 MMP 抑制剂的作用机制尚未得到充分了解，即人类蛋白质 TIMP-1 和来自绿茶的预防癌症的 EGCG。这两种具有治疗潜力的作用机制将进一步研究。之前的项目发现典型 TIMP 和 MMP 的高亲和力是由熵增益驱动的，很大程度上是配置性的。 N-TIMP-1 β-桶的主链在与 MMP-3 结合后被动员。为了更完整地了解与其结合相关的灵活性变化，核磁共振迁移率研究将扩展到 MMP-3 的游离态和 N-TIMP-1 结合态、侧链甲基以及热力学推论变得更加稳健的更长的时间尺度。 N-TIMP-1 的工程改造以增强治疗潜力，却忽略了其不良的生长因子活性。之前的项目在 TIMP 上发现了一个保守的表面，假设可以介导这种活动。该贴片将被诱变，以消除 N-TIMP-1 刺激细胞增殖的活性。将确定介导 TIMP-1 生长因子活性的信号通路和受体。绿茶中主要多酚 EGCG 的化学预防生物活性中发现了对少数 MMP 的抑制作用。将调查哪些 MMP 被 EGCG 抑制。将研究EGCG抑制MMP的新机制及其结合MMP的结构模式。当代治疗心血管和肺部疾病的药物靶点是 MMP-12。 MMP-12 对血管和肺的弹性蛋白成分以及弹性蛋白酶的弹性蛋白保留抑制剂（称为 a1-抗胰蛋白酶）具有独特的高活性。 MMP-12 对弹性蛋白和α1-抗胰蛋白酶的活性的序列决定因素尚不清楚，将进行探索。 MMP-12 中与 a1-抗胰蛋白酶相互作用的位点将通过 NMR 进行定位。弹性蛋白酶特异性将从 MMP-12 中改造出来，然后转移到 MMP-3 中。赋予 MMP-12 独特特异性的特征的发现应有助于设计更具选择性的抑制剂，并可能提示其他 MMP 特异性的决定因素。