Administrative Supplements to Support Undergraduate Summer Research Experiences

项目摘要 这项Mira提案旨在揭示金属蛋白酶的调节机制和底物。 ADAMTS(一种带有血栓反应蛋白基序的去整合素和金属蛋白酶)家族。这些蛋白酶被认为是 参与了许多生物过程，其中一些成员对人类 疾病。目前的研究主要集中在ADAMTS家族的两个成员，ADAMTS13和ADAMTS7。 项目1将研究ADAMTS13的变构调节，ADAMTS13是一种重要的凝血调节因子。我最近 发现ADAMTS13采用静止的闭合构象，并通过其变构激活 底物，Von Willebrand因子(VWF)。这项提议概述了一种研究变构如何 监管在ADAMTS13内维持，并因其与VWF的相互作用而被扰乱。利用功能强大、 动力学、结构和生物信息学分析，该项目旨在确定变构的关键结构元素 ADAMTS13中的监管。这项工作的结果将提高我们对遗传和后天的理解 ADAMTS13缺陷引起的凝血障碍。我还预计我们对ADAMTS13的调查 将作为研究其他ADAMTS蛋白水解酶调控的模型。项目2的重点是ADAMTS7，它 已知通过促进动脉粥样硬化和炎症而导致冠状动脉疾病。然而， ADAMTS7的底物靶点及其生化特性仍然知之甚少。之前的尝试是 解决这些研究问题导致了文献中相互矛盾的报告，可能是由于 可用于研究这种新型蛋白酶的试剂和工具。为了绕过这些挑战，我们将使用新颖的 蛋白质组学、酶学和蛋白质-蛋白质相互作用的方法来描述生物化学特性和 ADAMTS7的理想基板。新发现的底物将成为新的生化工具的基础 研究ADAMTS7的活性，对于未来的研究目标筛选小分子抑制剂是必要的 可能具有翻译应用的ADAMTS7。ADAMTS13和ADAMTS7的研究综述 酶的催化和调节为我们理解血栓和出血性疾病奠定了基础 心血管和炎症性疾病，这些结果可以用于研究其他ADAMTS 蛋白酶。更重要的是，Mira为首席调查员提供了一个难以置信的机会来招募和 指导来自不同背景的学生和其他实习生。

Project Summary This MIRA proposal aims to uncover the regulatory mechanisms and substrates for metalloproteases in the ADAMTS (A Disintegrin And Metalloprotease with ThromboSpondin motifs) family. These proteases are thought to be involved in many biological processes and some members are known to contribute to important human diseases. The current study focuses mainly on two members of the ADAMTS family, ADAMTS13 and ADAMTS7. Project 1 will investigate the allosteric regulation of ADAMTS13, an important regulator of blood clotting. I recently discovered that ADAMTS13 adopts a quiescent closed conformation and becomes allosterically activated by its substrate, Von Willebrand Factor (VWF). This proposal outlines a mechanistic approach to study how allosteric regulation is maintained within ADAMTS13 and disrupted by its interaction with VWF. Leveraging functional, kinetic, structural and bioinformatic analyses, this project sets to identify the key structural elements of allosteric regulation in ADAMTS13. The outcomes from this work will improve our understanding of inherited and acquired blood clotting disorders caused by defects in ADAMTS13. I also anticipate that our investigation of ADAMTS13 will serve as a model to study the regulation of other ADAMTS proteases. Project 2 focuses on ADAMTS7, which is known to contribute to coronary artery disease by promoting atherosclerosis and inflammation. However, the substrate targets of ADAMTS7 and its biochemical properties remain poorly understood. Previous attempts to address these research questions led to conflicting reports in the literature, likely due to the poor quality of reagents and tools available to study this novel protease. To circumvent these challenges, we will utilize novel methods in proteomics, enzymology, and protein-protein interactions to delineate biochemical properties and ideal substrates for ADAMTS7. Newly identified substrates will form the basis of novel biochemical tools to investigate ADAMTS7 activity and are necessary for future research goals to screen for small molecule inhibitors of ADAMTS7 that may have translational applications. Summarily, the study of ADAMTS13 and ADAMTS7 enzyme catalysis and regulation lay a foundation for our understanding of thrombotic and bleeding disorders and cardiovascular and inflammatory diseases, and these outcomes can be applied to study other ADAMTS proteases. More importantly, the MIRA offers the principal investigator an incredible opportunity to recruit and mentor students and other trainees from diverse backgrounds.