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，已经发明了新的药物，其诱导致病蛋白质与 一种叫做泛蛋白连接酶的酶，可以促进有问题的蛋白质的破坏。这种新药 模式是每年提供十亿美元的工业推动，以利用泛素连接酶来治疗各种人类疾病。 疾病这些努力中的大多数都利用了称为Cullin-RING连接酶（CRL）的酶家族。 在人类中有大约200个成员，CRL共同控制了大约20%的泛素依赖性蛋白。 细胞中的蛋白质降解。因此，了解这些酶是如何调节的是相当重要的。 引起了广大观众的兴趣。已知人类CRL与至少7种另外的酶合作， 我们称之为泛素携带酶（UCEs），它是促进CRL依赖性蛋白底物 降解CRL的控制被认为主要通过其可逆的 被一种叫做NEDD 8的蛋白质修饰。此外，它也被认为是UCE行为杂乱 在CRL-UCE交互水平上排除CRL监管的CRL。然而，CRL-UCE 活性CRL的最新结构强烈暗示了特异性。R 01赠款的初步结果 表明CRL-UCE特异性赋予这些对以异常快速泛素转移速率， 潜在地提供除neddylation之外的CRL功能的附加控制层。考虑到这些 根据观察，本行政补充旨在获得停流仪器来开发生物物理 蛋白质-蛋白质相互作用测定，以测量UCE之间蛋白质复合物形成的动力学 和CRL。从这些研究中产生的结果将有助于解释在一个机制水平上， 实现了特异性。