Coordination of chaperone interactions that dictate protein folding and trafficking

决定蛋白质折叠和运输的伴侣相互作用的协调

• 批准号: 10000953
• 负责人: Lars Plate
• 金额: $ 39.48万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000953
• 关键词: 3-Dimensional; Affect; Alzheimer' s Disease; Binding; Biology; Cells; Chemicals; Client; Complex; Coupled; Cretinism; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Defect; Dependence; Destinations; Disease; Ensure; Individual; Intervention; Lead; Location; Mass Spectrum Analysis; Mediating; Methodology; Molecular Chaperones; Molecular Conformation; Neurodegenerative Disorders; Organelles; Parkinson Disease; Pathway interactions; Process; Protein Dynamics; Protein Subunits; Proteins; Proteomics; Quality Control; Shapes; Structure; Therapeutic; Thyroglobulin; Thyroid Hormones; Time; Variant; insight; loss of function; mutant; novel strategies; premature; protein aggregation; protein folding; protein misfolding; protein transport; proteostasis; tool; trafficking

PROJECT SUMMARY Proteins fold into their 3-dimensional shape with the help of chaperones and other protein folding factors, which together comprise the proteostasis network (PN). During the protein quality control process, transient binding interactions between individual client proteins and proteostasis factors mediate folding into native functional structures, thereby ensuring trafficking to the correct cellular destination, or facilitating degradation of detrimental misfolded states. Consequently, imbalances in interactions between proteostasis factors and clients proteins result in quality control defects that lead to diverse protein misfolding diseases including highly prevalent neurodegenerative diseases such as Alzheimer’s and Parkinson’s Disease. The folding, maturation and trafficking of large multi-domain and multi-subunits proteins is a complex and highly client-specific process that depends on engaging the appropriate component of the PN at the correct time. Many proteostasis dependencies for individual client proteins, are known, but little is understood about the order of engagement, and whether correct sequential interactions are required for proper folding and trafficking. Understanding the coordination of the PN interaction with client proteins at an organelle- or cell-wide level will be crucial to determine the mechanisms of quality control defects that impact protein misfolding diseases. We utilize the power of chemical biology and quantitative proteomics tools to determine the interaction dynamics between disease-associated protein variants and the PN. This enables us to investigate the mechanism by which altered progression through the PN impacts protein quality control. To probe the progression of the client proteins through protein folding and trafficking pathways, we will establish new proteomics methodology to elucidate dynamically changing interactions both globally and in a time-dependent manner. We will examine the coordination requirement of different proteostasis pathways as they affect protein aggregation (thryroglobulin) and loss-of-function protein misfolding (cystic fibrosis transmembrance conductance regulator – CFTR). Aggregation of destabilized thyroglobulin variants, a thyroid hormone precursor, is a leading cause for congenital hypothyroidism, while misfolding and pre-mature degradation of CFTR variants is the primary cause of Cystic Fibrosis. In particular, we will assess how protein quality control of mutant variants of these proteins is impacted by altered PN interactions and how established and new approaches to manipulate proteostasis pathways can correct these quality control defects. Results from our studies will provide significant new insights into how the dynamics of protein folding and trafficking pathways can be manipulated therapeutically for disease intervention.

项目总结 蛋白质在伴侣蛋白和其他蛋白质折叠的帮助下折叠成三维形状 这些因素共同构成了蛋白质平衡网络(PN)。在蛋白质质量控制过程中 过程中，单个客户蛋白与蛋白稳定因子之间的瞬时结合相互作用 折叠成本地功能结构，从而确保运输到正确的细胞目的地，或者 促进有害的错误折叠状态的退化。因此，互动中的不平衡 蛋白质平衡因子和客户蛋白质之间的质量控制缺陷导致不同的 蛋白质错误折叠疾病，包括阿尔茨海默氏症等高度流行的神经退行性疾病 和帕金森氏症。大分子多结构域多亚单位的折叠、成熟和运输 蛋白质是一个复杂的、高度特定于客户的过程，它依赖于参与适当的 在正确的时间产生PN的分量。单个客户蛋白的许多蛋白质平衡依赖关系， 都是已知的，但对交战的顺序以及是否正确的顺序了解很少 相互作用是正确折叠和贩运所必需的。理解PN的协调 在细胞器或细胞水平上与客户蛋白质的相互作用将是决定 影响蛋白质错误折叠疾病的质量控制缺陷的机制。 我们利用化学生物学和定量蛋白质组学工具来确定 疾病相关蛋白变体与Pn之间的相互作用动力学。这使我们能够 研究通过PN改变进程影响蛋白质质量控制的机制。 为了探索客户蛋白质通过蛋白质折叠和运输途径的进展，我们将 建立新的蛋白质组学方法以阐明动态变化的全球和 以依赖于时间的方式。我们将研究不同蛋白质平衡的协调要求。 影响蛋白质聚集(甲状腺球蛋白)和功能丧失的蛋白质错误折叠的途径 (囊性纤维化跨膜电导调节剂-CFTR)。不稳定的聚集 甲状腺球蛋白变异体是一种甲状腺激素前体，是先天性甲状腺功能减退的主要原因， 而CFTR变异体的错误折叠和早熟降解是囊性纤维化的主要原因。在……里面 特别是，我们将评估这些蛋白质的突变变体的蛋白质质量控制是如何受到 改变的PN相互作用以及如何建立和新的方法来操纵蛋白平衡通路 可以纠正这些质量控制缺陷。我们的研究结果将提供重要的新见解 蛋白质折叠和运输途径的动力学如何在治疗上被操纵 疾病干预。