Decoding the molecular logic of TPR cochaperones

解读 TPR 共伴侣的分子逻辑

基本信息

批准号：
10463466
负责人：
Matthew Callahan
金额：
$ 3.95万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
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/10463466
关键词：
Adaptor Signaling Protein Affinity Affinity Chromatography Amino Acids Binding Biochemical Biological Assay Biology C-terminal Cells Cellular Stress Chemicals Client Complex Computer Models Development Disease Ensure Environment Family Family member Financial compensation Future Genetic Goals Health Heat shock proteins Heat-Shock Proteins 70 Human In Vitro Individual Lead Ligand Binding Ligands Light 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 temporal measurement tool ubiquitin-protein ligase

项目摘要

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.

项目概要/摘要四肽重复序列 (TPR) 辅助伴侣是一个多样化的接头蛋白家族，与热休克蛋白 (Hsp) 70 和 Hsp90 伴侣系统。这些模块化蛋白质由它们组成同名 TPR 结构域，介导与 Hsp 的复合物形成，以及酶促或支架可以帮助监护人招募客户或质量控制的域名。这些配件的重要性 TPR 共伴侣在广泛的应用中的意义证明了维持蛋白质稳态的功能从神经退行性疾病到癌症等一系列疾病。然而，我们对分子机制的理解支持这些蛋白质的底物识别的基础是不完整的。特别是，越来越多的证据 TPR 结构域可以招募超出其典型伴侣结合伙伴的蛋白质，其重要性目前尚不清楚这种替代途径的作用。另外，一般来说，建立底物是很困难的。考虑到蛋白质稳态内可能发生的补偿，TPR 辅助伴侣的关系遗传扰动后的网络。开发特异性抑制TPR的化学探针因此，具有高时间分辨率的共伴侣复合物是非常理想的。努力开发这样的通过了解 TPR 域的哪些分子特征可以大大增强工具的功能用于实现高亲和力和选择性结合。该提案的目标是开发一种化学品通过解码 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共伴侣及其底物。在我的第一个目标中，我将开发化学蛋白质组学工具来分析结合 TPR 共伴侣并测量底物关联响应不同刺激的变化。这些探针还将能够快速评估 TPR 抑制剂的特异性。在我的第二个目标中，我将完善预测评分功能，以全面识别自主的底物被 E3 连接酶 CHIP 识别。在探索 CHIP 与其相互作用的分子决定因素时底物，我还将确定使单个 TPR 共伴侣能够结合不同配体的特征来自其相关家庭成员。这项工作意义重大，因为它将提供对 TPR 共伴侣的生物学规则，并将作为未来底物发现的基石并探索该蛋白质家族的开发工作。拟议的研究还将提供培训非常适合我成为一名转化化学生物学家的目标的环境，有机会获得最先进技术的经验，同时培养我进行研究的能力独立。