Early Events in Protein Folding

蛋白质折叠的早期事件

• 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
• 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

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.

项目摘要/摘要 结构功能范式是一种强大的指导原则，是我们许多理解的基础 生物过程。然而，这张图中通常忽略的是蛋白质结构的灵活性。 这种灵活性对于蛋白质折叠到其天然活性结构是必需的。此外，蛋白质功能 需要随着时间的流逝来演变这种天然结构。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 该建议是阐明启用血凝素和M2质子通道的功能动力学 流感病毒感染，这是一个重要的公共卫生影响的问题。探索的机制 这项工作也与其他包膜病毒有关，尤其是艾滋病毒。更一般地，膜融合 质子通过蛋白质具有很高的基本利益，我们希望获得的见解 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的机制。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 在激光诱导的pH跳跃上开发了独特的方法基础，以启动融合过程和结构 特定的光谱方法来表征驱动膜的血凝素重折叠动力学 融合。该病毒蛋白是理解膜一般机制的原型 融合为无处不在的膜运输过程。 2）确定融合孔形成的机理。我们将测试血凝素的假设 跨膜结构域（TMD）和融合肽（FP）形成一种寡聚复合物，可以打开并稳定 融合孔。 3）确定主动门控质子转运的分子机制。这项工作将重点 流感M2质子通道，一个重要的模型离子通道。了解质子的运输 蛋白质通道对于许多基本生物学过程以及流感病毒的复制至关重要。 这些目标在智力上与能源景观概念联系在一起，并在操作上由方法论链接 在我们的实验室中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 确定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。 我们希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 膜蛋白和耦合的膜动力学。