Investigating the regulation and substrate processing of ADAMTS13 and ADAMTS7 metalloproteases.

研究 ADAMTS13 和 ADAMTS7 金属蛋白酶的调节和底物加工。

• 批准号: 10626794
• 负责人: Joshua Mutambuki Muia
• 金额: $ 33.35万
• 依托单位: OSU CENTER FOR HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626794
• 关键词: Address; Adopted; Allosteric Regulation; Atherosclerosis; Biochemical; Bioinformatics; Biological Process; Blood Coagulation Disorders; Blood coagulation; Cardiovascular Diseases; Catalysis; Coronary Arteriosclerosis; Defect; Development; Diagnosis; Disease; Disintegrins; Elements; Enzymatic Biochemistry; Enzymes; Family; Foundations; Goals; Inflammation; Inflammatory; Inherited; Investigation; Kinetics; Literature; Metalloproteases; Methods; Modeling; Molecular Conformation; Outcome; Peptide Hydrolases; Prevention; Principal Investigator; Property; Proteomics; Reagent; Regulation; Reporting; Research; Thrombosis; Thrombospondins; Work; biochemical tools; human disease; improved; member; novel; protein protein interaction; recruit; small molecule inhibitor; student mentoring; therapeutic target; tool; translational applications; von Willebrand Factor

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.

项目摘要 该MIRA提案旨在揭示金属蛋白酶的调节机制和底物 ADAMTS（具有血小板传播基序的分解蛋白和金属蛋白酶）家族。这些蛋白酶被认为 参与许多生物过程，有些成员有助于重要人类 疾病。当前的研究主要关注Adamts家族的两个成员Adamts13和Adamts7。 项目1将研究ADAMTS13的变构调节，ADAMTS13是血液凝结的重要调节剂。我 最近发现Adamts13采用静止的封闭构象并变成了变构 由其底物Von Willebrand因子（VWF）激活。该提议概述了一种研究的方法 变构调节如何在ADAMTS13中维持并因其与VWF的相互作用而破坏。利用 功能，动力学，结构和生物信息学分析，该项目旨在识别关键结构元素 ADAMTS13中的变构调节。这项工作的结果将改善我们对继承的理解 并获得了由ADAMTS13缺陷引起的血液凝结障碍。我也预计我们对 ADAMTS13将作为研究其他ADAMTS蛋白酶调节的模型。项目2关注 ADAMTS7，已知通过促进动脉粥样硬化和 炎。但是，ADAMTS7及其生化特性的底物靶标保持较差 理解。以前解决这些研究问题的尝试导致文献中的报告相互矛盾， 可能是由于可用于研究这种新型蛋白酶的试剂和工具的质量差。绕过这些 挑战，我们将利用蛋白质组学，酶学和蛋白质 - 蛋白质相互作用的新方法来描述 ADAMTS7的生化特性和理想底物。新确定的底物将构成 研究ADAMTS7活动的新型生化工具，对于将来的研究目标是筛查的必要条件 对于可能具有翻译应用的ADAMTS7的小分子抑制剂。总而言之，研究 ADAMTS13和ADAMTS7酶催化和调节为我们了解血栓的基础 以及出血性疾病以及心血管和炎症性疾病，这些结果可以应用于 研究其他ADAMTS蛋白酶。更重要的是，MIRA为主要研究人员提供了令人难以置信的 有机会招募和指导来自不同背景的学生和其他学员。