Structural Characterization of Prion Isoforms in Multiple TSE Diseases

多种 TSE 疾病中朊病毒亚型的结构特征

• 批准号: 8053786
• 负责人: Michele Ann McGuirl
• 金额: $ 27.56万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8053786
• 关键词: Affect; Amyloid Proteins; Bovine Spongiform Encephalopathy; Brain; C-terminal; Cattle; Characteristics; Chemicals; Creutzfeldt-Jakob Syndrome; Detection; Deuterium; Disease; Electron Microscopy; Electrons; Environment; Family; Fatal Familial Insomnia; Gerstmann-Straussler-Scheinker Disease; Glean; Goals; Health; Human; Hydrogen; Individual; Inherited; Kuru; Label; Length; Mammals; Maps; Mass Spectrum Analysis; Membrane; Membrane Proteins; Methods; Modeling; Molecular Conformation; Mutation; N-terminal; Nature; Neurodegenerative Disorders; Peptide Fragments; PrPSc Proteins; Prion Diseases; Prions; Property; Protein Isoforms; Protein Subunits; Proteins; Publishing; Recombinants; Relative (related person); Reporter; Research; Resolution; Scrapie; Series; Sheep; Site; Spectrum Analysis; Spin Labels; Structure; Surveys; Techniques; Testing; Tissues; Variant; Vertebral column; Work; X-Ray Crystallography; base; chronic wasting disease of elk and deer; crosslink; disease transmission; interest; mutant; prevent; protein function; protein misfolding; research study; solute; synuclein; tau Proteins; trend

DESCRIPTION (provided by applicant): The goal of this research is to elucidate the structure of the prion protein in its aggregated isoforms, and to elucidate the relationship between isoform structure and different types of prion diseases. Prion protein is the causative agent of many fatal neurodegenerative diseases of mammals. These diseases are characterized by the conversion of normal, monomeric prion protein (PrPC) to a misfolded conformational state that aggregates and accumulates as fibrillar plaques in the brain. Transmission of the disease can occur when one mammal ingests the infected tissue of another. When exposed to an infectious aggregate, normal PrPC takes on the misfolded conformation and the fibrils are thus propagated. Any treatment of a prion disease must prevent or abrogate misfolding/aggregation while not affecting normal PrP function. Hence, it is important to understand the structural differences between the PrP isofoms. High resolution structures of PrPC from many mammals have been determined by both NMR and X-ray crystallography. In contrast, the insoluble, fibrillar nature of the infectious form has hampered the elucidation of its structural details. However, numerous biophysical and spectroscopic studies suggest that the misfolded form contains a much higher proportion of ¿-sheet than normal PrP, which is mostly a-helical/random coil. This work will elucidate the nature of the PrP subunit interactions in several forms of recombinant PrP by using two biophysical techniques: site directed spin labeling (SDSL) combined with electron paramagnetic spectroscopy (EPR) and site directed crosslinking combined with mass spectrometry (MS). The EPR experiments provide information on protein backbone dynamics, secondary, tertiary, and quaternary structure, distances between labeled residues, and the mobility of the label itself. The crosslinking experiments will confirm the structural information gleaned from EPR. PUBLIC HEALTH RELEVANCE The elucidation of the details of the structure of prion fibrils will be of great value in developing treatments for prion diseases, which are fatal neurodegenerative diseases of mammals. These include Creutzfeldt-Jakob disease (CJD) and variant CJD, Gerstmann- Straussler-Scheinker disease, Fatal Familial Insomnia, and kuru (humans), scrapie (sheep), chronic wasting disease (elk, deer), and mad cow disease (cattle). Treatment of any prion disease must prevent or abrogate fibril formation while not affecting normal prion function; hence, it is important to understand the structural differences among the different prion isofoms.

描述（由申请人提供）：本研究的目的是阐明朊病毒蛋白在其聚集亚型中的结构，并阐明亚型结构与不同类型朊病毒疾病之间的关系。朊蛋白是哺乳动物许多致命性神经退行性疾病的致病因子。这些疾病的特征在于正常的单体朊病毒蛋白（PrPC）转化为错误折叠的构象状态，其聚集并积累为脑中的纤维状斑块。当一种哺乳动物摄入另一种哺乳动物的受感染组织时，就会发生疾病的传播。当暴露于感染性聚集体时，正常PrPC呈现错误折叠的构象，因此原纤维繁殖。朊病毒疾病的任何治疗都必须防止或消除错误折叠/聚集，同时不影响正常的PrP功能。因此，重要的是要了解PrP异构体之间的结构差异。高分辨率结构的PrPC从许多哺乳动物已经确定了NMR和X射线晶体学。相反，感染性形式的不溶性纤维状性质阻碍了其结构细节的阐明。然而，许多生物物理学和光谱学研究表明，错误折叠的形式包含比正常PrP高得多的<$-片层比例，其主要是α-螺旋/无规卷曲。这项工作将阐明的性质PrP亚基的相互作用，在几种形式的重组PrP通过使用两种生物物理技术：定向自旋标记（SDSL）结合电子顺磁光谱（EPR）和定向交联结合质谱（MS）。EPR实验提供了关于蛋白质骨架动力学、二级、三级和四级结构、标记残基之间的距离以及标记本身的流动性的信息。交联实验将证实EPR收集的结构信息。公共卫生相关性朊病毒原纤维结构细节的阐明将在开发朊病毒疾病的治疗方法中具有重要价值，朊病毒疾病是哺乳动物的致命神经变性疾病。这些疾病包括克雅氏病（CJD）和变异型CJD、Gerstmann-Straussler-Scheinker病、致命性家族性传染病和库鲁病（人类）、羊瘙痒病（绵羊）、慢性消耗性疾病（麋鹿、鹿）和疯牛病（牛）。任何朊病毒疾病的治疗都必须在不影响正常朊病毒功能的同时预防或消除原纤维的形成;因此，了解不同朊病毒亚型之间的结构差异是很重要的。