STRUCTURE AND GENESIS OF TAU FILAMENTS

申请人发现了一种全新的、强大的 一种将tau蛋白聚合成类似于直丝的细丝的方法 在阿尔茨海默病的神经病理学中发现的细丝。 诱导剂是膜磷脂和脂肪酸， 已知在体内起信号转导分子的作用。 所得 长丝是规则的并且可大量获得。 通过分析 这些细丝的结构和起源，他们将解决重要的 关于tau纤维形成和神经退行性疾病的问题。 哪些膜相关成分能够刺激tau蛋白 自缔合和聚合 结果是否 构效关系指向已知信号的参与 丝生物发生的转导机制？ 的作用是什么 磷酸化对tau蛋白自组装和细胞内稳定性的影响？ 四目 是为了解决这些问题而提出的。 1)tau纤维的过程 将形成量化。 使用一种新的组装试验，他们将 量化tau蛋白丝形成的速率、程度和配体依赖性 体外 2)丝形成所需的tau蛋白结构特征 将被确定。 这些研究将阐明结构要求 tau蛋白聚合和配体结合位点的位置 可用于开发高亲和力的治疗剂， 诊断应用。 3)翻译后的影响 将评估对τ聚合动力学的改变。 这些研究 将阐明磷酸化和糖化对tau蛋白丝的影响 阵 4)聚合对tau结构的影响将是 测定 使用圆二色谱偏振光谱法 和水动力测量，将确定tau是否采用 在长丝形成时具有限定的结构。 结果将具有重要意义 对开发能够 预防，和能够检测的死前诊断剂， 纤维病理学的形成。

DESCRIPTION The applicants have discovered an entirely new and powerful method of polymerizing tau protein into filaments resembling the straight filaments found in the neurofibrillary pathology of Alzheimer's disease. The inducing agents are membrane phospholipids and fatty acids that are known to function as signal transduction molecules in vivo. The resultant filaments are regular, and available in large quantities. By analyzing the structure and genesis of these filaments, they will address important questions regarding tau filament formation and neurodegenerative disease. Which membrane-associated components are capable of stimulating tau self-association and polymerization? Does the resultant structure-activity-relationship point toward the involvement of known signal transduction mechanisms in filament biogenesis? What is the role of phosphorylation on tau self-assembly and intracellular stability? Four Aims are proposed to address these questions. 1) The process of tau filament formation will be quantified. Using a novel assembly assay, they will quantify the rate, extent, and ligand dependence of tau filament formation in vitro. 2) The structural features on tau required for filament formation will be determined. These studies will clarify the structural requirements of tau polymerization and the location of a ligand-binding site potentially useful for development of high-affinity agents for therapeutic and diagnostic applications. 3) The influence of post-translational modification on tau polymerization kinetics will be assessed. These studies will clarify the impact of phosphorylation and glycation on tau filament formation. 4) The effects of polymerization on tau structure will be determined. Using a combination of circular dichroism spectropolarimetry and hydrodynamic measurements, it will be determined whether tau adopts a defined structure upon filament formation. The results will have important implications for the feasibility of developing therapeutic agents capable of preventing, and premortem diagnostic agents capable of detecting, the formation of fibrillar pathology.