Synthetically generated mammalian prions

合成哺乳动物朊病毒

• 批准号: 9088526
• 负责人: JIYAN MA
• 金额: $ 58.84万
• 依托单位: VAN ANDEL RESEARCH INSTITUTE
• 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 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9088526
• 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; 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（由申请人提供）：朊病毒是一种基于蛋白质的感染因子，可引起一组称为传染性海绵状脑病（或朊病毒疾病）的神经退行性疾病。尽管多年的研究，朊病毒传播的分子机制仍然不清楚。最近开发的蛋白质错误折叠循环扩增（PMCA）协议产生高度感染性的朊病毒从细菌表达的重组朊病毒蛋白的存在下，定义明确的辅因子提供了独特的机会，探测朊病毒传播的分子基础。在这个多PI应用中，我们建议使用结合体外重组朊病毒的产生、动物生物测定和生物物理/结构表征的综合方法来（i）阐明脂质在朊病毒感染性中的作用，（i）严格检验强朊病毒传播屏障可以通过系列适应性构象变化废除的假设，和（iii）建立一种新的重组朊病毒增殖方案，以产生大量的感染性朊病毒，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朊病毒的结构和生物学特性，有望为朊病毒传播机制提供有价值的见解。此外，鉴于最近发现的“朊病毒样”传播的有序蛋白聚集体在神经退行性疾病，如阿尔茨海默氏症和帕金森氏病，从我们的研究结果也应该有重要的意义，了解这些更常见的神经退行性疾病的致病过程。