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

基本信息

批准号：
10672931
负责人：
Lars Plate
金额：
$ 39.59万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2025-06-30
项目状态：
未结题

项目摘要

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.

项目摘要蛋白质借助伴侣和其他蛋白质折叠折叠成3维形状因素，共同构成了蛋白质的网络（PN）。在蛋白质质量控制期间过程，单个客户蛋白和蛋白质抗体因子之间的瞬时结合相互作用介导折叠成本地功能结构，从而确保运输到正确的蜂窝目的地或促进有害错误折叠状态的退化。因此，互动失衡在蛋白质的因素和客户蛋白之间，蛋白质会导致质量控制缺陷，从而导致潜水蛋白质错误折叠疾病，包括高度普遍的神经退行性疾病，例如阿尔茨海默氏症和帕金森氏病。大型多域和多婚姻的折叠，成熟和贩运蛋白质是一个复杂且高度客户特定的过程，取决于参与适当的过程 PN的组件在正确的时间。许多对单个客户蛋白的蛋白质依赖性依赖性，已知，但对参与顺序以及是否正确顺序几乎没有理解适当的折叠和贩运需要相互作用。了解PN的协调在细胞器或细胞水平上与客户蛋白的相互作用对于确定质量控制缺陷的机制会影响蛋白质错误折叠疾病。我们利用化学生物学和定量蛋白质组学工具的力量来确定疾病相关蛋白质变体与PN之间的相互作用动力学。这使我们能够研究通过PN改变进展会影响蛋白质质量控制的机制。为了探测客户蛋白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论，以阐明全球和以时间依赖的方式。我们将检查不同蛋白质的协调要求途径在影响蛋白质聚集（thryroglobulin）和功能丧失蛋白质折叠时的途径（囊性纤维化透射电导调节器 - CFTR）。不稳定的聚集甲状腺球蛋白变体是一种甲状腺龙骨前体，是先天性甲状腺功能减退症的主要原因，虽然CFTR变体的错误折叠和预性降解是囊性纤维化的主要原因。在特别是，我们将评估这些蛋白质突变变体的蛋白质质量控制如何受到改变了PN相互作用，以及如何操纵蛋白质途径的建立和新方法可以纠正这些质量控制缺陷。我们研究的结果将提供重要的新见解如何对蛋白质折叠和运输途径的动力学进行热操作疾病干预。