MMP-2 in Periodontal Disease and Oral Cancer

描述：MMP-2是基质金属蛋白酶家族的一员 基质金属蛋白酶（MMPs），它们一起切割广泛的组织成分。虽然这 MMPs的特性是正常发育和组织的有益特征， 适应，不受控制的MMP-2活性已与 炎症性疾病，如牙周病和关节炎，以及肿瘤 扩张和转移。该应用程序旨在开发化合物， 其特异性抑制MMP-2活性。 由于MMP-2对分子的切割只有在MMP-2和MMP-2之间存在结合时才发生， 酶和底物分子，MMP-2结合的特定机制 将研究其主要胶原底物。在一次合作努力中， 分子生物学和蛋白质结构分析方法将应用于 首先通过随机筛选胶原蛋白上的特异性MMP-2结合位点， 肽库和定位胶原蛋白上的功能肽序列。到 确定MMP-2上精确的胶原结合位点残基，核磁共振， 共振研究将用于分析MMP-2胶原结合结构域 (CBD)与合成肽复合，模拟CBD结合位点， 胶原鉴定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测试，并通过分析 CBD中的位点特异性突变。 一旦胶原蛋白和MMP-2上的精确结合位点被确定， 将开发能够抑制全长天然MMP-2的分子， 通过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MMP-2配体结合和 活性测定和用MMP-2表达细胞的实验。 拟议的研究应确定特异性结合位点相互作用 MMP-2与其主要胶原基质之间的关系，并探索新的策略， 在炎症性疾病和癌症中抑制MMP-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.