Molecular Mechanisms of Prion and Amyloid Propagation

朊病毒和淀粉样蛋白传播的分子机制

• 批准号: 8667470
• 负责人: Christopher P Jaroniec
• 金额: $ 28.98万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8667470
• 关键词: Amino Acids; Amyloid; Amyloid Fibrils; Architecture; Atomic Force Microscopy; Biochemical; Biology; Bovine Spongiform Encephalopathy; C-terminal; Cattle; Characteristics; Chronic Wasting Disease; Creutzfeldt-Jakob Syndrome; Data; Deer; Deuterium; Disease; Exhibits; Fingerprint; Generations; Hamsters; Hereditary Cerebral Amyloid Angiopathy; Human; Hydrogen; In Vitro; Individual; Infectious Agent; Link; Magic; Mammals; Measures; Mesocricetus auratus; Methods; Molecular; Molecular Conformation; Mus; Mutation; N-terminal; NMR Spectroscopy; Nature; Neurodegenerative Disorders; Neurologic; PrP; PrP variant; PrPSc Proteins; Prion Diseases; Prions; Protein Conformation; Proteins; Public Health; Publishing; Relaxation; Research; Resolution; Role; Scheme; Scrapie; Seeds; Sheep; Side; Site; Solutions; Specificity; Structural Models; Structure; Techniques; Transmission Electron Microscopy; Validation; Variant; Vertebral column; Wild Type Mouse; Work; Yeasts; base; conformational conversion; conformer; design; disease phenotype; flexibility; fungus; in vitro Model; insight; meetings; methionylmethionine; monomer; mutant; novel; prevent; prion hypothesis; prion-based; protein aggregate; restraint; scaffold; solid state nuclear magnetic resonance; transmission process

DESCRIPTION (provided by applicant): The prion concept, according to which proteins alone can be infectious, represents a new paradigm in biology. Such protein-only infectious agents are believed to be responsible for transmissible spongiform encephalopathies, a group of fatal neurodegenerative disorders that include Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy in cattle. Mammalian prions propagate by a mechanism involving the conformational conversion of a largely a-helical cellular prion protein, PrPC, to misfolded, ¿-rich amyloid-like aggregates, PrPSc. More recently, the prion hypothesis has been extended further to include the phenomenon of protein conformation-based inheritance in yeast and other fungi. Particularly puzzling features of such conformational infectivity and inheritance are the existence of so-called prion strains linked to discrete disease phenotypes within the same host species, and their role in transmissibility barriers which frequently prevent efficient prion transmission between different species. While recent studies indicate that prion strains originate from the ability of proteins to misfold into multiple distinct conformers in forming self-propagating amyloid aggregates, molecular-level understanding of these phenomena has been hampered by the paucity of high-resolution structural data. In this project we aim to advance the understanding of fundamental aspects of protein conformation-based inheritance and infectivity by providing detailed insights into the molecular mechanisms and structural basis of amyloid propagation and transmissibility barriers for a non-infectious Y145Stop PrP variant (PrP23-144). This C-truncated PrP mutant is associated with hereditary cerebral amyloid angiopathy in humans and exhibits in vitro some of the most fundamental aspects of PrP propagation including the phenomena of prion strains and species barriers. The specific aims focus on the determination and validation of a detailed structural model for human PrP23-144 amyloid aggregates, and elucidation of the structural differences related to the emergence of distinct PrP23-144 amyloid strains with unique transmission characteristics (i.e., seeding specificities) linked to mutations of two critical amino acid residues near the C-terminus of PrP23-144. Modern multidimensional solid-state NMR spectroscopy will be the primary experimental structural technique employed in the study. In addition, we will use hydrogen/deuterium exchange solution NMR methods, atomic force microscopy and tilted-beam transmission electron microscopy.

描述（由适用提供）：根据其单独蛋白质可以具有感染性的prion概念，代表了生物学的新范式。据信这种仅蛋白质传染剂是导致可传播的海绵状脑病，这是一群致命的神经退行性疾病，其中包括人类和牛在牛中的牛皮刺激性脑病。哺乳动物的prions通过一种机制传播，涉及大部分A螺旋细胞蛋白PRPC的构象转化，以折叠折叠，�-富丽淀粉样蛋白的聚集体Prpsc，PRPSC。最近，王室假设已进一步扩展，以包括酵母和其他真菌中基于蛋白质会议的遗传的现象。这种构象感染和遗传性的拼图特征特别是存在与同一宿主物种中离散疾病表型相关的所谓的prion菌株，以及它们在传播障碍中的作用，这些菌株经常阻止不同物种之间有效的prion传播。尽管最近的研究表明，素菌株源于蛋白质在形成自传播的淀粉样蛋白聚集体中将蛋白错误折叠成多种不同的构象体的能力，但分子水平对这些现象的理解受到高分辨率结构数据的匮乏的阻碍。在该项目中，我们旨在通过提供对淀粉样蛋白机制的详细见解和淀粉样蛋白传播的分子机制和结构基础的详细见解，以促进对基于蛋白质构象的遗传和感染的基本方面的理解。该C截断的PRP突变体与人类的遗传性脑淀粉样蛋白血管病有关，并在体外表现出PRP传播的一些最基本方面，包括主要菌株和物种障碍的现象。该具体目的侧重于确定和验证人类PRP23-144淀粉样蛋白聚集体的详细结构模型，并阐明与与独特的传输特性（即，播种规格）相连的prp23-144淀粉样蛋白差异的不同质量差异的不同PRP23临近prp的突变的prp23 prp的突变的结构差异。现代的多维固态NMR光谱将是研究中采用的主要实验结构技术。此外，我们将使用氢/氘交换溶液NMR方法，原子力显微镜和倾斜梁透射电子显微镜。