Synthetically generated mammalian prions

合成哺乳动物朊病毒

• 批准号: 8605252
• 负责人: JIYAN MA
• 金额: $ 52.33万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-15 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8605252
• 关键词: Affect; Alzheimer' s Disease; Amino Acids; Amyloid Fibrils; Animals; Atomic Force Microscopy; Biochemical; Biological; Biological Assay; Bovine Spongiform Encephalopathy; Brain; Creutzfeldt-Jakob Syndrome; Cultured Cells; Data; Deuterium; Disease Outbreaks; Endopeptidase K; Foundations; Funding; Future; Generations; Hamsters; Health; Human; Hydrogen; In Vitro; Individual; Infectious Agent; Lipids; Mass Spectrum Analysis; Methods; Molecular; Molecular Conformation; Molecular Probes; Molecular Structure; Mus; Neurodegenerative Disorders; Parkinson Disease; Play; Prion Diseases; Prions; Process; Property; Protein Conformation; Proteins; Protocols documentation; Reaction; Recombinants; Reporting; Research; Resistance; Resolution; Role; Shaker potassium channel; Site-Directed Mutagenesis; Sonication; Structure; Techniques; Testing; Variant; amyloid structure; base; cofactor; conformer; design; insight; large scale production; microbial; neuropathology; novel; prion-based; prion-like; protein aggregate; protein misfolding cyclic amplification; public health relevance; transmission process

DESCRIPTION (provided by applicant): Prion is a protein-based infectious agent that causes a group of neurodegenerative disorders known as transmissible spongiform encephalopathies (or prion diseases). Despite years of researches, the molecular mechanism of prion transmission remains unclear. The recently developed protein misfolding cyclic amplification (PMCA) protocol for generating highly infectious prions from bacterially expressed recombinant prion protein in the presence of well-defined cofactors offers unique opportunities to probe the molecular basis of prion transmission. In this multiple PI application, we propose to use an integrated approach combining generation of recombinant prions in vitro, animal bioassay, and biophysical/structural characterization to (i) elucidate the role of lipids in prion infectivity, (i) rigorously test the hypothesis that a strong prion transmission barrier can be abrogated by serial adaptive conformational changes, and (iii) establish a novel recombinant prion propagation protocol to generate large quantities of infectious prions and, thus, facilitate future structural studies at a higher resolution level. These studies, which integrate structural and biological characterization of recombinant prions, are expected to provide valuable insights into the mechanism of prion transmission. Moreover, given the recent discoveries of a "prion-like" propagation of ordered protein aggregates in neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases, the findings from our studies should also have important implications for understanding the pathogenic process in these more common neurodegenerative diseases.

描述(申请人提供)：Prion是一种基于蛋白质的感染性物质，可导致一组神经退行性疾病，称为传染性海绵状脑病(或Prion疾病)。尽管进行了多年的研究，但PrP传递的分子机制仍不清楚。最近发展起来的蛋白质错折叠循环扩增(PMCA)方法，可以在明确的辅因子存在的情况下，从细菌表达的重组普恩蛋白中产生高感染性的普恩病毒，为探索普恩病毒传播的分子基础提供了独特的机会。在这种多重PI应用中，我们建议使用一种综合的方法，结合体外产生重组Prion、动物生物测定和生物物理/结构表征来(I)阐明类脂在Prion感染性中的作用，(I)严格检验强Prion传递屏障可以通过一系列适应性构象变化来消除的假设，以及(Iii)建立一种新的重组Prion繁殖方案来产生大量的传染性Prion，从而促进未来在更高分辨率水平上的结构研究。这些研究整合了重组Pron的结构和生物学特性，有望为Pron的传播机制提供有价值的见解。此外，鉴于最近在阿尔茨海默氏症和帕金森氏病等神经退行性疾病中发现了有序的蛋白质聚集体的“普里恩”繁殖，我们的研究结果对于理解这些更常见的神经退行性疾病的致病过程也应该具有重要的意义。