Degenerative and Dementing Diseases of Aging

描述(由申请人提供)：总体来说，在四个科学项目和四个核心中，我们建议研究导致神经退化的A？PrP和tau蛋白。Prion导致阿尔茨海默病和其他神经退行性疾病，包括额颞叶痴呆、帕金森氏病和ALS的发现，为破解这些疾病的发病机制和开发新的治疗方法打开了新的研究策略。理解所有普恩的基础是确定特定蛋白质在成为普恩时所经历的结构转变。在这个P01更新申请中，我们计划利用我们和其他人发表的新数据，这些数据认为A？和tau，像PrP一样，可以获得自繁殖的构象；因此，它们是普恩。在每种情况下，这些可供选择的构象都富含？片状结构，并容易在淀粉样原纤维中聚合，这些纤维经常凝结成斑块或缠结。在项目1中，我们建议使用我们创建的双基因小鼠来研究Aüpron菌株的特性，开发能够检测Aü和tau蛋白的培养细胞生物检测方法，使用表达BV PrP或嵌合的Hu/BVPrP转基因的双基因小鼠来研究人类(Hu)PrP Prion，并确定表达BVPrP的培养细胞是否能够支持人类Pron菌株的复制。在项目2中，我们建议继续研究由重组PrP形成的PrPSc的结构，利用启动无锚定PrPSc Prion复制的89聚体片段，以继续结构研究 重点是自然发生的突变，并启动tau蛋白的结构研究。在项目3中，我们建议通过溶液和固态核磁共振来确定Aüpron的结构；使用对全局和局部构象敏感的多个探针来监测Aü组装；创建增强或抑制淀粉样蛋白起始、延伸和碎裂的单个步骤的含有硫代酰胺的多肽和折叠分子；以及合成一系列交联剂来探测A？、PrP和tau蛋白中Lys和Arg残基之间的距离分布。在项目4中，我们建议将集成结构建模应用于蛋白质自组装，从而促进基于不同类型的稀疏、噪声、歧义和不连贯数据的多个结构状态的同时建模。

DESCRIPTION (provided by applicant): Overall In four scientific projects and four cores, we propose to study Aß PrP, and tau prions causing neurodegeneration. The discovery that prions cause Alzheimer's disease and other neurodegenerative diseases including frontotemporal dementias, Parkinson's disease, and ALS opens new research strategies for deciphering the pathogenesis of these illnesses and developing novel therapeutic approaches. Fundamental to understanding all prions is defining the structural transition that a particular protein undergoes when it becomes a prion. In this P01 renewal application, we plan to exploit new data published by us and others contending that Aß and tau, like PrP, can acquire conformations that are self-propagating; thus, they are prions. In each case, these alternative conformations are enriched for ß-sheet structure and readily polymerize in amyloid fibrils that often condense into plaques r tangles. In Project 1, we propose to study the properties of strains of Aß prions using bigenic mice that we created, to develop cultured cell bioassays that can detect Aß and tau prions, to investigate human (Hu) PrP prions using bigenic mice expressing bank vole (BV) PrP or chimeric Hu/BVPrP transgenes, and to determine if cultured cells expressing BVPrP can support the replication of human prion strains. In Project 2, we propose to continue structural studies of PrPSc formed from recombinant PrP utilizing an 89-mer fragment that initiated replication of anchorless PrPSc prions, to continue structural studies of Aß with emphasis on naturally occurring mutants, and to initiate structural studies of tau prions. In Project 3, we propose to determine the structure of Aß prions by solution and solid-state NMR; to monitor Aß assembly using multiple probes that are sensitive to both global and local conformations; to create thioamide- containing peptides and foldamers that enhance or inhibit individual steps of amyloid initiation, elongation, and fragmentation; and to synthesize a series of crosslinking reagents to probe the distribution of distances between Lys and Arg residues in Aß, PrP and tau prions. In Project 4, we propose to adapt and apply integrative structural modeling to protein self-assembly, thus facilitating simultaneous modeling of multiple structural states based on sparse, noisy, ambiguous and incoherent data of different kinds.