How ubiquitin-carrying enzymes contribute to ubiquitin ligase specificity

泛素携带酶如何促进泛素连接酶特异性

• 批准号: 10783452
• 负责人: Gary L. Kleiger
• 金额: $ 8.61万
• 依托单位: UNIVERSITY OF NEVADA LAS VEGAS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10783452
• 关键词: Administrative Supplement; Biological Assay; Biophysics; Cells; Cullin Proteins; Disease; Endowment; Enzyme Interaction; Enzymes; Family; Grant; Human; Industry; Investigation; Kinetics; Ligase; Measures; Modality; Modification; Molecular; Pharmaceutical Preparations; Protac; Proteins; Regulation; Research; Specificity; Structure; Therapeutic; Ubiquitin; feeding; human disease; insight; instrument; interest; invention; member; novel therapeutics; protein complex; protein degradation; protein protein interaction; ubiquitin ligase

In recent years novel drugs have been invented that induce proximity between a disease-causing protein and an enzyme called a ubiquitin ligase that promotes the destruction of the problematic protein. This new drug modality is feeding a billion dollar per year industry push to employ ubiquitin ligases to treat various human diseases. Most of these efforts have been utilizing a family of enzymes called the Cullin-RING ligases (CRLs). With some 200 members in humans, the CRLs collectively control approximately 20 % of ubiquitin-dependent protein degradation in cells. As such, an appreciation for how these enzymes are regulated is of considerable interest to a very wide audience. Human CRLs are known to partner with at least 7 additional enzymes, which we refer to as ubiquitin-carrying enzymes (UCEs), that are required to promote CRL-dependent protein substrate degradation. The control of CRLs is believed to be determined predominantly through their reversible modification with a protein called NEDD8. Furthermore, it also was believed that UCEs act promiscuously towards CRLs which would preclude CRL regulation at the level of CRL-UCE interaction. However, CRL-UCE specificity is strongly implied by the recent structure of an active CRL. Preliminary results from our R01 grant indicate that CRL-UCE specificity endows these pairs with exceptionally rapid rates of ubiquitin transfer, potentially providing an additional layer of control of CRL function beyond neddylation. In consideration of these observations, this administrative supplement seeks to obtain a stopped-flow instrument to develop biophysical protein-protein interaction assays to measure the kinetics of formation of the protein complexes between UCEs and CRLs. The results generated from these studies will help explain on a mechanistic level how CRL-UCE specificity is achieved.

近年来，人们发明了新的药物来诱导致病蛋白和蛋白质之间的接近