Structural Determinants of Mammalian Prion Aggregation

哺乳动物朊病毒聚集的结构决定因素

• 批准号: 10239229
• 负责人: Calina Glynn
• 金额: $ 1.27万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-09-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10239229
• 关键词: Affect; Amyloid; Amyloidosis; Animal Diseases; Behavioral Symptoms; Biochemical; Biology; Caring; Characteristics; Chemicals; Code; Communicable Diseases; Contracts; Cryoelectron Microscopy; Crystallization; Crystallography; Dementia; Diagnosis; Disease; Disease Outbreaks; Disease susceptibility; Electrons; Endopeptidase K; Environmental Health; Event; Exhibits; FDA approved; Fiber; Filament; Genetic; Genetic Polymorphism; Goals; Human; Infectious Agent; Length; Link; Mammals; Methods; Micro Electron Diffraction; Microtus; Molecular; Morphology; Neurodegenerative Disorders; Organism; Pathology; Patients; Pattern; Peptide Hydrolases; Peptides; Play; Polymorph; Population; PrP; PrP sequence; Predisposition; Prion Diseases; Prions; Property; Proteins; Proteolysis; Reaction; Recombinants; Resistance; Resolution; Role; Sampling; Sequence Alignment; Stretching; Structure; Therapeutic; Variant; base; beta pleated sheet; disease phenotype; disease transmission; disorder prevention; experimental study; frontier; interest; non-genetic; particle; pathogen; prevent; psychiatric symptom; response; structural biology; therapeutic development; therapy design; therapy development; transmission process; wild-type PrP

Project Summary: Prion diseases are neurodegenerative disorders that pose a threat to human and environmental health. These illnesses have genetic and nongenetic causes, can be environmentally acquired, and have the ability to remain infectious in the absence of a host organism for extended periods. Aggregation of misfolded prion protein (PrPSc) correlates with disease, but the mechanism behind aggregation is not fully understood, gravely hindering the development of therapeutics to prevent fibrillation and sickness. The β2α2 loop of mammalian prion protein has previously been demonstrated to be a key region implicated in disease transmission. A crystal structure from a nine-residue segment encoding the β2α2 loop of the bank vole prion has demonstrated structural characteristics and stability characteristic of full-length prion fibers. Building on this structure, structures of PrP aggregates that convey information on stability and infectivity will be pursued. To achieve this goal, ordered aggregates of PrP segments that have been described to play a role in disease transmission in various species with a range of prion disease susceptibility will be biochemically and structurally characterized. Constructs incorporating disease modulating regions will be recombinantly expressed, purified, and fibrillized to assess stability, proteinase K resistance, and fiber morphologies. These properties will be compared against those observed in fibrils derived from diseased animals. A combination of crystallography and single particle Cryo-EM will be used to determine the atomic arrangement of each misfolded PrP. These aims will be achieved through the application of frontier methods in macromolecular crystallography including electron micro-diffraction (MicroED). The resulting structures will provide a molecular explanation for a nearly three- hundred-year-old mystery in prion biology and protein pathology and will help distinguish infectious from non- infectious amyloids, revealing structural code for prion transmission barriers. !

项目总结： Pron病是一种神经退行性疾病，对人类和环境健康构成威胁。这些 疾病有遗传和非遗传原因，可以在环境中获得，并有能力保持 在长时间没有寄主生物体的情况下具有传染性。错误折叠的Pron蛋白的聚集 (PrPSc)与疾病相关，但聚集背后的机制尚未完全了解，严重 阻碍预防纤颤和疾病的治疗方法的发展。哺乳动物的β-2-α-2环 Prion蛋白此前已被证明是参与疾病传播的关键区域。一个 编码银行田鼠β2α2环的9个残基片段的晶体结构已经证明 全长Pron纤维的结构特征和稳定性特征。在这个结构的基础上， 将寻求PrP聚集体的结构，以传达关于稳定性和传染性的信息。要做到这一点 目标，PrP片段的有序聚集，已被描述为在疾病传播中发挥作用 将从生化和结构上描述具有一系列Pron疾病易感性的各种物种。 含有疾病调节区的构建物将被重组表达、纯化和纤维连接以 评估稳定性、对蛋白酶K的抵抗力和纤维形态。这些属性将与 在患病动物的纤维中观察到的那些。结晶学和单粒子的结合 冷冻电子显微镜将被用来确定每个错误折叠的PrP的原子排列。这些目标将是 通过在包括电子在内的大分子结晶学中应用前沿方法实现的 微电子衍射(MicroED)。由此产生的结构将为近三个- 在普里子生物学和蛋白质病理学中的百年谜团，将有助于区分感染性和非传染性 传染性淀粉样蛋白，揭示了Pron传递障碍的结构代码。 好了！