MMP-2 in Periodontal Disease and Oral Cancer

描述：基质金属蛋白酶-2是基质金属蛋白酶家族的一员。 (MMPs)，它们共同切割广泛的组织成分。虽然这件事 MMPs的特性是正常发育和组织的有益特征 适应，不受控制的基质金属蛋白酶-2活性与 炎症性疾病，如牙周病和关节炎，以及肿瘤扩张和转移。这个应用程序旨在开发化合物， 它能特异性抑制基质金属蛋白酶-2的活性。 因为只有当分子与基质金属蛋白酶-2结合时才会发生分子切割 酶和底物分子，基质金属蛋白酶-2结合的具体机制 它的主要胶原蛋白底物将被研究。在合作的努力下， 分子生物学和蛋白质结构分析方法将应用于 首先通过筛选一个随机的结合位点来鉴定胶原上的特异性基质金属蛋白酶-2结合位点 多肽文库，并定位胶原蛋白上的功能多肽序列。至 确定基质金属蛋白酶-2上精确的胶原结合部位残基，核磁 共振研究将用于分析基质金属蛋白酶-2胶原结合结构域 (CBD)与模拟CBD结合部位的合成肽形成络合物 胶原蛋白。识别出的位点和氨基酸的特异性将是 在竞争性配体结合试验中进行了测试，并通过分析 CBD中的定点突变。 一旦确定了胶原和基质金属蛋白酶-2的精确结合部位， 将开发出能够抑制全长天然基质金属蛋白酶-2的分子 竞争底物结合和取代结合位点的活性 CBD上的残留物。这将在基质金属蛋白酶-2配体结合和 活性检测，并用表达基质金属蛋白酶-2的细胞进行实验。 拟议的研究应定义特定的结合部位相互作用 基质金属蛋白酶-2及其主要胶原底物之间的关系，并探索一种新的治疗策略 抑制炎症性疾病和癌症中的基质金属蛋白酶-2 底物结合。

DESCRIPTION: MMP-2 is a member of the family of matrix metalloproteinases (MMPs), which together cleave a broad range of tissue components. While this property of the MMPs is a beneficial feature of normal development and tissue adaptation, uncontrolled MMP-2 activity has been strongly associated with inflammatory diseases, such as periodontal disease and arthritis, and tumor expansion and metastasis. This application is designed to develop compounds, which specifically inhibit MMP-2 activity. Since cleavage of molecules by MMP-2 occurs only if there is binding between the enzyme and substrate molecules, the specific mechanism by which MMP-2 binds its main collagen substrates will be investigated. In a collaborative effort, molecular biology and protein structural analysis methods will be applied to first identify specific MMP-2 binding sites on collagen by screening a random peptide library and mapping the functional peptide sequences on collagen. To identify the precise collagen binding site residues on MMP-2, nuclear magnetic resonance studies will be used to analyze the MMP-2 collagen binding domain (CBD) complexed with synthetic peptides, which mimic the CBD binding sites on collagen. The specificity of the identified sites and amino acids will be tested in competitive ligand binding assays and by analyzing the effects of site-specific mutations in the CBD. Once the precise binding sites on both collagen and MMP-2 are defined, small molecules will be developed that can inhibit the full-length native MMP-2 activity by competing for substrate binding and by substituting binding site residues on the CBD. This will be accomplished in both MMP-2 ligand binding and activity assays, and in experiments with MMP-2 expressing cells. The proposed studies should define the specific binding site interactions between MMP-2 and its main collagen substrate and explore a new strategy to inhibit MMP-2 in inflammatory diseases and cancer based on competition for substrate binding.