Early Events in Protein Folding

蛋白质折叠的早期事件

基本信息

批准号：
10217148
负责人：
RICHARD BRIAN DYER
金额：
$ 35.64万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-06-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10217148
关键词：
Address Antiviral Agents Binding Biological Process Capsid Proteins Cell physiology Cells Cessation of life Collaborations Complex Coupled Coupling Cytosol Dehydration Disease Disease Outbreaks Drug resistance Economic Burden Endosomes Equilibrium Event Evolution Fluorescence Fluorescence Resonance Energy Transfer Free Energy Goals HIV Hemagglutinin Immunologics Infection Influenza Influenza Hemagglutinin Ion Channel Isotopes Kinetics Lasers Length Link Lipid Bilayers Lipids Maps Mediating Membrane Membrane Fluidity Membrane Fusion Membrane Proteins Methodology Methods Modeling Molecular Molecular Conformation Molecular Machines Motion Nature Pathogenesis Pathologic Processes Peptides Pharmaceutical Preparations Phase Process Protein Dynamics Proteins Protons Public Health Reaction Research Ribonucleoproteins Role Series Shapes Solvents Specificity Spectrum Analysis Structure System Tertiary Protein Structure Testing Time Transmembrane Domain Transmembrane Transport Transport Process Viral Viral Proteins Virus Diseases Virus Replication Water Work base drug development flexibility influenza M2 influenza infection influenzavirus insight interest molecular dynamics mutant neglect novel strategies pandemic disease peptide structure prevent protein folding protein function protein structure response simulation single-molecule FRET time use transmission process virus envelope

项目摘要

Project Summary/Abstract The structure-function paradigm is a powerful guiding principle that underlies much of our understanding of biological processes. What is often neglected in this picture, however, is the flexibility of protein structures. This flexibility is necessary for a protein to fold to its native, active structure. Furthermore, protein function requires evolution of this native structure with time. Therefore, the dynamics of the protein structure and associated solvent water provide the critical connection between structure and function. The overall goal of this proposal is to elucidate the functional dynamics of hemagglutinin and M2 proton channel that enable influenza virus infection, a problem with significant public health implications. The mechanisms explored in this work are also relevant to other enveloped viruses, in particular HIV. More generally, membrane fusion and proton transport through proteins are of high fundamental interest and we expect the insight gained in these specific studies will contribute to the understanding of a broad range of related systems. We plan to pursue three specific aims: 1) Determine the mechanism of hemagglutinin mediated membrane fusion. We will test a new model for protein mediated membrane fusion that is based on molecular dynamics simulations of this process. We have developed unique methodology base on a laser induced pH jump to initiate the fusion process, and structure specific spectroscopic methods to characterize the hemagglutinin refolding dynamics that drive membrane fusion. This viral protein serves as an archetype for understanding the general mechanism of membrane fusion as a ubiquitous membrane transport process. 2) Determine the mechanism of fusion pore formation. We will test the hypothesis that the hemagglutinin trans-membrane domain (TMD) and fusion peptide (FP) form an oligomeric complex that opens and stabilizes the fusion pore. 3) Determine the molecular mechanism of actively gated proton transport. This work will on a focus on the influenza M2 proton channel, an important model ion channel. Understanding transport of protons through protein channels is critical to many essential biological processes as well as replication of the influenza virus. These aims are linked intellectually by energy landscape concepts and operationally by the methodology developed in our lab for studying both protein and membrane dynamics. Our unique approach will allow us to identify specific protein motions involved in protein mediated membrane fusion and proton channel activation. We expect this work to provide important new insight into the factors that shape the energy landscape of membrane proteins and the coupled membrane dynamics.

项目概要/摘要结构-功能范式是一个强有力的指导原则，是我们许多理解的基础的生物过程。然而，这张图中经常被忽视的是蛋白质结构的灵活性。这种灵活性对于蛋白质折叠成其天然的活性结构是必要的。此外，蛋白质功能需要这种原生结构随着时间的推移而演变。因此，蛋白质结构的动力学和相关的溶剂水提供了结构和功能之间的关键联系。总体目标为该提案旨在阐明血凝素和 M2 质子通道的功能动力学，从而使流感病毒感染是一个对公共卫生产生重大影响的问题。探索的机制这项工作也与其他包膜病毒有关，特别是艾滋病毒。更一般地，膜融合和质子通过蛋白质的运输具有很高的根本意义，我们期望在这些具体研究将有助于理解广泛的相关系统。我们计划追求三个具体目标： 1) 确定血凝素介导的膜融合机制。我们将测试一个新模型蛋白质介导的膜融合基于该过程的分子动力学模拟。我们有开发了基于激光诱导 pH 跃变的独特方法来启动融合过程和结构用于表征驱动膜的血凝素重折叠动力学的特定光谱方法融合。这种病毒蛋白可以作为理解膜一般机制的原型融合作为普遍存在的膜运输过程。 2)确定熔孔形成机理。我们将检验血凝素的假设跨膜结构域 (TMD) 和融合肽 (FP) 形成寡聚复合物，可打开并稳定融合孔。 3）确定主动门控质子传输的分子机制。这项工作将重点关注流感M2质子通道，一种重要的模型离子通道。了解质子通过蛋白质通道对于许多重要的生物过程以及流感病毒的复制至关重要。这些目标在智力上通过能源景观概念联系起来，在操作上通过方法论联系起来我们的实验室开发用于研究蛋白质和膜动力学。我们独特的方法将使我们能够识别参与蛋白质介导的膜融合和质子通道激活的特定蛋白质运动。我们期望这项工作能够为塑造能源格局的因素提供重要的新见解。膜蛋白和耦合膜动力学。