CHEMICAL APPROACHES FOR ACTIVITY BASED PROTEOMICS

该提案的目的是开发和利用新的 用于基于活性的蛋白质组学研究的化学探针， 了解水解酶在肿瘤转移中的作用。与 后基因组时代的迅速到来，新的战略， 需要对蛋白质进行分析。到目前为止，蛋白质组学的工作主要是 他们关注的是记录蛋白质水平的变化，而不是活动。 根据蛋白质活性的变化对其进行分析的能力 这将大大加快蛋白质功能的分配， 确定潜在的药物靶点。本文所描述的工作 该提案将侧重于设计、合成和测试新型活性物质， 定点探针，记录动态的表达和功能， 整个酶家族。在本申请中，PI将使用合成 化学，生物化学，分子和细胞生物学技术， 目标1）鉴定支持或阻碍肿瘤转移的蛋白酶，2） 与肿瘤相关的蛋白水解活性的细胞外“热点”成像 细胞迁移和侵袭，以及3）开发基于活性的选择性探针 基质金属蛋白酶家族的成员。通过这些研究， PI希望澄清在许多水解酶中， 实际上参与蛋白水解事件的重要进展， 疾病这些酶将反过来成为药物治疗的有价值的靶点。 旨在抑制癌症转移的努力。

The objective of this proposal is to develop and utilize novel chemical probes for activity-based proteomics investigations aimed at understanding the role that hydrolytic enzymes play in tumor metastasis. With the post-genome era rapidly approaching, new strategies for the functional analysis of proteins are needed. To date, proteomics efforts have primarily been concerned with recording variations in protein level rather than activity. The ability to profile classes of proteins based on changes in their activity would greatly accelerate both the assignment of protein function and the identification of potential pharmaceutical targets. The work described in this proposal will focus on designing, synthesizing, and testing novel active site-directed probes that record dynamics in the expression and function of entire enzyme families. In this application, the PIs will use synthetic chemistry, biochemistry, and molecular and cell biology techniques towards the goals of 1) identifying proteases that support or impede tumor metastasis, 2) imaging extracellular "hot spots" of proteolytic activity associated with tumor cell migration and invasion, and 3) developing activity-based probes selective for the matrix metalloproteinase family of enzymes. Through these studies the PI hopes to clarify which, of the many, hydrolytic enzymes implicated in cancer actually participate in proteolytic events important for the progression of the disease. The enzymes will in turn represent valuable targets for pharmaceutical efforts aimed at suppressing cancer metastasis.