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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目摘要将几个功能单元（称为域）组合到单个多肽链中是一个常见创造生物复杂性的进化策略。由此产生的多域蛋白是在所有蛋白质组织中都普遍，并执行必要的细胞功能。但是，功能增加这些大蛋白的复杂性使它们折叠成本地功能结构。相比之下对于许多较小的蛋白质或单个域，多域蛋白很容易折叠，并且潜在的细胞毒性聚集。在单元格中，几个因素确保有效折叠。折叠开始共同翻译时，核糖体仍然合成多肽。分子伴侣开始一旦核糖体出现，它就会与新生的多域蛋白相互作用。共同翻译折叠和伴侣相互作用被认为对有效的多域至关重要蛋白质折叠。但是，这些过程在分子水平上的定义仍然很差，因为它是在技术上具有挑战性地研究它们。该项目的目的是定义共同翻译的原则折叠和伴侣功能，以更好地了解复杂多域蛋白如何鲁棒他们的功能结构。我们使用了单分子生物物理学和活细胞的组合实现这一目标的实验。使用光学镊子，我们正在研究新生的多域蛋白在单分子水平上。单个分子的操作是非常适合解决新生蛋白的复杂折叠途径，阐明核糖体和分子伴侣到折叠过程，并确定如何共同翻译折叠和蛋白质合成耦合并调节以确保稳健的结果。这些详细说明在检测共同翻译事件的活细胞中实验的实验补充了体外研究在多域蛋白中。蛋白质错误折叠和聚集，蛋白质合成和伴侣功能的下降是许多与衰老有关的疾病的标志。我们的研究可能最终提供了发现新颖的治疗策略来治疗其中一些的机械基础疾病。