SDSL-EPR Study of Intermediate Filament Structure

描述(申请人提供)：中间丝(IF)是存在于每个细胞中的三大类细胞骨架之一，也是含量较丰富的细胞蛋白质之一。IF蛋白的突变导致至少85种不同的人类疾病，包括许多蛋白质聚集性疾病。然而，这些蛋白质的不同寻常的物理性质排除了确定晶体结构的可能性。正因为如此，我们没有深入了解正常IF功能的机制，也没有深入了解突变是如何致病的。在这个提案中，我们将使用生物物理学的方法来定义IF结构，目的是阐明IF的功能，以及IF结构在IF类中的保守程度。这反过来将为了解IF蛋白，特别是那些具有蛋白质聚集表型的IF蛋白的发病机制打开大门。我们已经组建了一支由几项技术组成的团队：CW SDSL EPR、脉冲场SDSL EPR、计算建模、高分辨率瞬变电子显微镜(低温和STEM)和X射线结晶学。

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.