Folding and Chaperone Interactions of Multi-domain Proteins

多结构域蛋白质的折叠和分子伴侣相互作用

• 批准号: 10662086
• 负责人: Christian Kaiser
• 金额: $ 20.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662086
• 关键词: Aging; Alzheimer' s Disease; Bacteria; Binding; Biological; Biology; Biophysics; Brain; Cell physiology; Cells; Codon Nucleotides; Communication; Complement; Complex; Contracts; Coupled; Cultured Cells; Cytosol; Detection; Disease; Ensure; Eukaryota; Event; Future; Geometry; Goals; In Vitro; Individual; Length; Link; Malignant Neoplasms; Measurement; Measures; Mechanics; Methods; Molecular; Molecular Chaperones; Muscle; Neurodegenerative Disorders; Neurons; Nutrient; Outcome; Parkinson Disease; Pathway interactions; Peptides; Peptidyltransferase; Process; Property; Protein Biosynthesis; Proteins; Proteome; Regulation; Resolution; Ribosomes; Role; Shapes; Structure; Tertiary Protein Structure; Translations; Variant; Work; Yeasts; attenuation; cytotoxic; defined contribution; experimental study; in vivo; insight; laser tweezer; mechanical force; novel therapeutic intervention; overexpression; polypeptide; protein aggregation; protein folding; protein misfolding; proteostasis; reconstitution; ribosome profiling; single molecule; three dimensional structure

Project Summary Combining several functional units, termed domains, into a single polypeptide chain is a common evolutionary strategy for creating biological complexity. The resulting multi-domain proteins are prevalent in all proteomes and carry out essential cellular functions. However, the increased functional complexity of these large proteins complicates their folding into native functional structures. In contrast to many smaller proteins or individual domains, multi-domain proteins are prone to misfolding and potentially cytotoxic aggregation. In the cell, several factors ensure efficient folding. Folding begins co- translationally, while the ribosome still synthesizes the polypeptide. Molecular chaperones begin to interact with the nascent multi-domain protein as soon as it emerges from the ribosome. Co- translational folding and chaperone interactions are recognized as crucial for efficient multi-domain protein folding. However, these processes remain poorly defined at the molecular level, because it is technically challenging to study them. The goal of this project is to define principles of co-translational folding and chaperone function to better understand how complex multi-domain proteins robustly reach their functional structures. We are using a combination of single-molecule biophysics and live-cell experiments to accomplish this goal. With optical tweezers, we are studying the folding pathways of nascent multi-domain proteins at the single-molecule level. Manipulation of individual molecules is ideally suited to resolve complex folding pathways of nascent proteins, elucidate the contributions of the ribosome and molecular chaperones to the folding process, and determine how co-translational folding and protein synthesis are coupled and regulated to ensure robust outcomes. These detailed in vitro studies are complemented by experiments in live cells that detect co-translational folding events in multi-domain proteins. Protein misfolding and aggregation, misregulation of protein synthesis and decline of chaperone function are hallmarks of many aging-related diseases. Our studies may ultimately provide a mechanistic basis for discovering novel therapeutic strategies to treat some of these diseases.

项目概要 将多个功能单元（称为结构域）组合成单个多肽链是一种常见的方法 创造生物复杂性的进化策略。由此产生的多结构域蛋白质是 普遍存在于所有蛋白质组中并执行重要的细胞功能。然而，增加的功能 这些大蛋白质的复杂性使它们折叠成天然功能结构变得复杂。相比之下 对于许多较小的蛋白质或单个结构域，多结构域蛋白质容易发生错误折叠和 潜在的细胞毒性聚集。在细胞中，有几个因素可确保有效折叠。折叠开始合作 翻译时，核糖体仍然合成多肽。分子伴侣开始 一旦新生的多结构域蛋白从核糖体中出现，它就会与它相互作用。共同 平移折叠和伴侣相互作用被认为对于高效的多域至关重要 蛋白质折叠。然而，这些过程在分子水平上仍然没有得到很好的定义，因为它是 研究它们在技术上具有挑战性。该项目的目标是定义协同翻译的原则 折叠和伴侣功能，以更好地了解复杂的多结构域蛋白质如何稳健地到达 他们的功能结构。我们正在结合单分子生物物理学和活细胞 为实现这一目标而进行的实验。使用光镊，我们正在研究折叠路径 单分子水平上的新生多结构域蛋白。对单个分子的操纵是 非常适合解决新生蛋白质的复杂折叠途径，阐明 核糖体和分子伴侣参与折叠过程，并确定如何共翻译 折叠和蛋白质合成相互耦合和调节，以确保稳健的结果。这些详细说明在 活细胞实验对体外研究进行了补充，检测共翻译折叠事件 在多结构域蛋白质中。蛋白质错误折叠和聚集、蛋白质合成失调和 伴侣功能的下降是许多与衰老相关的疾病的标志。我们的研究最终可能 为发现治疗其中一些疾病的新治疗策略提供机制基础 疾病。