Decoding the molecular logic of TPR cochaperones

解读 TPR 共伴侣的分子逻辑

• 批准号: 10710162
• 负责人: Matthew Callahan
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10710162
• 关键词: Adaptor Signaling Protein; Affinity; Affinity Chromatography; Amino Acids; Binding; Biochemical; Biological Assay; Biology; C-terminal; Cells; Cellular Stress; Chemicals; Client; Compensation; Complex; Computer Models; Development; Disease; Ensure; Environment; Family; Family member; Future; Genetic; Goals; Health; Heat shock proteins; Heat-Shock Proteins 70; Human; In Vitro; Individual; Ligand Binding; Ligands; Logic; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Modeling; Molecular; Molecular Chaperones; Nerve Degeneration; Neurodegenerative Disorders; Pathway interactions; Peptides; Play; Post-Translational Protein Processing; Protein Family; Proteins; Proteome; Proteomics; Quality Control; Rapid screening; Regulation; Reproducibility; Research; Role; Running; Sampling; Specificity; Stimulus; Structure-Activity Relationship; System; Techniques; Testing; Training; Update; Validation; Work; experience; experimental study; human disease; improved; inhibitor; insight; novel; protein aminoacid sequence; protein expression; proteostasis; recruit; response; scaffold; screening; skills; temporal measurement; tool; ubiquitin-protein ligase

PROJECT SUMMARY / ABSTRACT Tetratricopeptide repeat (TPR) cochaperones are a diverse family of adaptor proteins that cooperate with the heat shock protein (Hsp) 70 and Hsp90 chaperone systems. These modular proteins are composed of their eponymous TPR domain, which mediates complex formation with the Hsps, and an enzymatic or scaffolding domain that can aid the chaperone in client recruitment or quality control. The importance of these accessory functionalities in maintaining protein homeostasis is evidenced by the implication of TPR cochaperones in a wide range of diseases from neurodegeneration to cancer. However, our understanding of the molecular mechanisms that underpin substrate recognition by these proteins is incomplete. In particular, there is increasing evidence that TPR domains can recruit proteins beyond their canonical chaperone binding partners, and the significance of this alternative pathway is currently unknown. In addition, it is generally difficult to establish substrate relationships for TPR cochaperones given the compensation that can occur within the protein homeostasis network following genetic perturbations. Development of chemical probes that can specifically inhibit TPR cochaperone complexes with high temporal resolution is therefore highly desirable. The effort to develop such tools would be greatly augmented by an understanding of which molecular features of TPR domains can be exploited to achieve high affinity and selective binding. The objective of this proposal is to develop a chemical toolkit that helps solve both of these problems by decoding the molecular logic of interactions between TPR cochaperones and their substrates. In my first aim, I will develop chemical proteomic tools to profile the binding of TPR cochaperones and measure changes in substrate association in response to different stimuli. These probes will also enable the specificity of TPR inhibitors to be assessed in a rapid fashion. In my second aim, I will refine a predictive scoring function in order to comprehensively identify substrates that are autonomously recognized by the E3 ligase CHIP. In exploring the molecular determinants of CHIP's interactions with its substrates, I will also identify features that enable an individual TPR cochaperone to bind ligands that are distinct from its related family members. This work is significant because it will provide fundamental insights into the rules governing the biology of TPR cochaperones, and will serve as the bedrock for future substrate discovery and probe development efforts across this protein family. The proposed studies will also provide a training environment that is ideally suited to my goal of becoming a translational chemical biologist, with opportunities to gain experience with state-of-the-art techniques while also cultivating my ability to conduct research independently.

项目摘要 /摘要 四肽重复（TPR）对伴侣是一个多元化的衔接蛋白家族，与 热休克蛋白（HSP）70和HSP90伴侣系统。这些模块化蛋白由它们的 同名TPR域，介导与HSP的复合物形成以及酶促或脚手架 可以帮助伴侣招聘或质量控制的域。这些配件的重要性 TPR联酮在宽阔的范围内的影响证明了维持蛋白质稳态的功能 从神经变性到癌症的疾病范围。但是，我们对分子机制的理解 这些蛋白质的基础底物识别是不完整的。特别是有越来越多的证据 TPR领域可以募集蛋白质超出其规范的伴侣结合伙伴，并且具有重要意义 目前未知的替代途径。另外，通常很难建立底物 鉴于蛋白质稳态内可能发生的补偿，TPR联合酮的关系 遗传扰动后的网络。可以特异性抑制TPR的化学探针的开发 因此，具有较高时间分辨率的联合酮配合物是非常需要的。开发这样的努力 了解TPR域的哪些分子特征可以极大地增强工具 利用以实现高亲和力和选择性结合。该建议的目的是开发一种化学物质 通过解码TPR之间相互作用的分子逻辑来帮助解决这两个问题的工具包 联酮及其底物。在我的第一个目标中，我将开发化学蛋白质组学工具来介绍绑定 TPR联酮的响应不同刺激的底物关联的变化。这些 探针还将使TPR抑制剂的特异性快速评估。在我的第二个目标中，我 将完善一个预测评分函数，以全面识别自主的基板 被E3连接酶芯片识别。在探索芯片相互作用的分子决定因素时 底物，我还将确定能够启用单个TPR cochaperone的功能，以绑定不同的配体 来自其相关家庭成员。这项工作很重要，因为它将提供对 管理TPR Cochaperones生物学的规则，并将作为未来底物发现的基石 并调查了该蛋白质家族的开发工作。拟议的研究还将提供培训 理想的环境适合我成为翻译化学生物学家的目标，并有机会 在最先进的技术中获得经验，同时还培养我进行研究的能力 独立。