Structures and Activities of Prions and Prion Proteins

朊病毒和朊病毒蛋白的结构和活性

• 批准号: 7964304
• 负责人: BYRON CAUGHEY
• 金额: $ 130万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7964304
• 关键词: Amyloid Fibrils; Binding; Biochemical; Bovine Spongiform Encephalopathy; Chronic Wasting Disease; Circular Dichroism; Complement; Complement 1q; Complex; Creutzfeldt-Jakob Syndrome; Data; Detection; Deuterium; Disease; Field Flow Fractionation; Fluorescence; Goals; Hydrogen; Infectious Agent; Left; Mass Spectrum Analysis; Membrane; Methodology; Methods; Molecular; Nature; Neurodegenerative Disorders; Nuclear Magnetic Resonance; Pathogenesis; Pentosan Polysulfate; Prion Diseases; Prions; Proteins; Recombinants; Scrapie; Structure; Therapeutic; Time; Work; disease transmission; flexibility; improved; particle; prion hypothesis; prion seeds; research study; three dimensional structure; transmission process

Accomplishments for FY2009: 1) We have shown for the first time that mammalian prions that cause a transmissible spongiform encephalopathy (TSE or prion disease) can be generated solely from bacterially expressed recombinant prion protein. This discovery provides the most compelling evidence so far for the potential protein-only prion hypothesis for the nature of the TSE infectious agent. However, the data leave open the likelihood that other molecules strongly enhance the infectious titers of pathological forms of prion protein. 2) We have extended our characterization of the structure of the flexible amino-terminal domain of PrP when bound to pentosan polysulfate, one of the most potent known anti-TSE therapeutic compounds. Nuclear magnetic resonance, fluorescence and circular dichroism experiments have refined our new three dimensional structure of the PrP:pentosan polysulfate complex. 3)We have refined our understanding of the smallest and most infectious prion particles using improved field-flow fractionation and prion detection methodologies. 4) We have pursued new methods for characterizing and visualizing the pathogenic form of prion protein when bound to membranes. 5) We have detected TSE strain-dependent differences in the interactions between prion protein and C1q, a complement factor. 6) We have use hydrogen-deuterium exchange and mass spectrometry to determined that infectious prion-seeded amyloid fibrils of bacterially expressed recombinant prion protein have a different structure than spontaneously formed (unseeded) fibrils. This work provided strong biochemical evidence of the distinct seeding capacity of infectious prions.

2009财年的成就：1）我们首次表明，引起可传播的海绵状脑病（TSE或PRION病）的哺乳动物prions可以仅由细菌表达的重组prion蛋白产生。这一发现为迄今为止的最引人注目的证据提供了针对TSE感染剂性质的潜在唯一蛋白质prion假说。但是，数据离开开放的可能性是其他分子强烈增强病毒蛋白质形式的传染性滴度。 2）当与Pentosan多硫酸盐（最有效的已知抗TSE治疗化合物之一）结合时，我们扩展了PRP柔性氨基末端结构域的结构的表征。核磁共振，荧光和圆形二分性实验已经完善了我们的PRP：Pentosan多硫酸盐复合物的新三维结构。 3）我们使用改进的场合分馏和prion检测方法来完善对最小，最感染性的prion颗粒的理解。 4）我们采用了新方法来表征和可视化与膜时的病毒蛋白的致病形式。 5）我们已经检测到TSE菌株依赖性差异在Prion蛋白与C1Q之间的相互作用（一种补体因子）之间。 6）我们已经使用氢交换和质谱法来确定细菌表达的重组prion蛋白的感染性亲蛋白种子淀粉样蛋白的结构与自发形成（未种子）原纤维的结构不同。这项工作提供了有力的生化证据，证明了感染性王室的独特播种能力。