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: 10202661
• 负责人: Lars Plate
• 金额: $ 39.59万
• 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/10202661
• 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.

项目摘要 蛋白质在分子伴侣和其他蛋白质折叠的帮助下折叠成三维形状 这些因素共同构成蛋白质稳态网络（PN）。在蛋白质质量控制过程中 过程，个别客户蛋白质和蛋白质稳态因子之间的瞬时结合相互作用介导 折叠成天然功能结构，从而确保运输到正确的细胞目的地，或 促进有害的错误折叠状态的降解。因此，相互作用的不平衡 蛋白质稳定因子和客户蛋白质之间的差异导致质量控制缺陷， 蛋白质错误折叠疾病，包括高度流行的神经变性疾病 和帕金森氏症大的多结构域和多亚基的折叠、成熟和运输 蛋白质是一个复杂的和高度客户特定的过程，取决于从事适当的 在正确的时间点上的PN分量。许多蛋白质稳定依赖于单个客户蛋白， 已知的，但很少有人了解的顺序，以及是否正确的顺序， 需要相互作用以进行适当的折叠和运输。理解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 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的力量来确定 疾病相关蛋白变体和PN之间的相互作用动力学。这使我们能够 研究通过PN改变进展影响蛋白质质量控制的机制。 为了探索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，以阐明全球范围内动态变化的相互作用， 以时间依赖的方式。我们将研究不同的蛋白酶抑制的协调要求 途径，因为它们影响蛋白质聚集（甲状腺球蛋白）和功能丧失蛋白质错误折叠 （囊性纤维化跨膜传导调节因子- CFTR）。不稳定的聚集 甲状腺球蛋白变体是甲状腺激素前体，是先天性甲状腺功能减退症的主要原因， 而CFTR变体的错误折叠和过早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的突变变体的蛋白质质量控制如何受到以下因素的影响： 改变的PN相互作用以及如何建立和新的方法来操纵蛋白抑制途径 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 如何在治疗上操纵蛋白质折叠和运输途径的动力学， 疾病干预。