SDSL-EPR Study of Intermediate Filament Structure

中间丝结构的SDSL-EPR研究

• 批准号: 8236802
• 负责人: PAUL Gillespie FITZGERALD
• 金额: $ 38.43万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8236802
• 关键词: Behavior; Biology; Cells; Computer Simulation; Cytoskeleton; Data; Databases; Disease; Electron Spin Resonance Spectroscopy; Filament; Funding; Goals; Head; Human; Intermediate Filament Proteins; Intermediate Filaments; Keratin; Knowledge; Mechanical Stress; Mediating; Metabolic stress; Molecular Chaperones; Molecular Structure; Mutation; Pathogenesis; Pathologic; Phenotype; Phosphorylation; Physiologic pulse; Point Mutation; Process; Property; Proteins; Resolution; Site; Spin Labels; Stretching; Structure; Tail; Techniques; Technology; Time; Transmission Electron Microscopy; Vimentin; X-Ray Crystallography; base; cellular targeting; cytotoxicity; dimer; disease-causing mutation; human disease; insight; member; mutant; physical property; protein aggregation; protein misfolding; retinal rods

DESCRIPTION (provided by applicant): Intermediate Filaments (IFs) are one of three major classes of cytoskeleton found in every cell, and one of the more abundant cellular proteins. Mutations in IF proteins are responsible for at least 85 different human diseases, including many protein aggregation diseases. However, the unusual physical properties of these proteins have precluded determination of crystal structure. Because of this we have no in-depth mechanistic understanding of normal IF function, nor how mutations are pathogenic. In this proposal we will use biophysical approaches to define IF structure with a goal of elucidating IF function, and the degree to which IF structure is conserved across IF classes. This in turn will open the door to understanding pathogenesis among IF proteins, particularly those with protein aggregation phenotypes. We have assembled a team of several technologies: CW SDSL EPR, Pulsed Field SDSL EPR, Computational Modeling, High resolution TEM (Cryo- and STEM) and X Ray Crystallography. PUBLIC HEALTH RELEVANCE: Intermediate Filaments (IFs) are a major component of every vertebrate cell. IFs help cells resist mechanical and metabolic stresses, and have been implicated in more than 85 human diseases. However, we have no mechanistic understanding of IF biology because we lack structure for IFs. Moreover the same properties that make IFs stable, also contribute to their potential to cause proteins aggregation diseases. We will solve normal IF structure, so that we can understand the mechanisms of normal IF function, thus laying the groundwork for understanding how specific mutations are pathogenic.

描述（由申请人提供）：中间细丝（IFS）是每个细胞中发现的三个主要类细胞骨架之一，也是一种更丰富的细胞蛋白。 IF蛋白质中的突变至少导致85种不同的人类疾病，包括许多蛋白质聚集疾病。但是，这些蛋白质的异常物理特性无法确定晶体结构。因此，我们对正常IF的功能没有深入的机理理解，也没有致病性。在此提案中，我们将使用生物物理方法来定义结构是否具有阐明功能的目标，以及在跨越类别中保存结构的程度。反过来，这将为理解IF蛋白质（尤其是具有蛋白质聚集表型的蛋白质的发病机理）打开大门。我们组装了一支由多种技术组成的团队：CW SDSL EPR，脉冲场SDSL EPR，计算建模，高分辨率TEM（冷冻和STEM）和X射线晶体学。 公共卫生相关性：中间细丝（IFS）是每个脊椎动物细胞的主要组成部分。 IFS有助于细胞抵抗机械和代谢应激，并与85种以上的人类疾病有关。但是，我们对IF生物学没有机械理解，因为我们缺乏IFS结构。此外，使IF稳定的相同特性也有助于它们引起蛋白质聚集疾病的潜力。我们将解决正常的结构，以便我们可以理解正常机制，从而为理解特定突变的致病性奠定了基础。