STRUCTURE AND GENESIS OF TAU FILAMENTS

申请者发现了一种全新的、强大的 将tau蛋白聚合成类似直丝的方法 在阿尔茨海默病的神经纤维性病理中发现的细丝。 诱导剂是膜磷脂和脂肪酸，它们是 已知在体内作为信号转导分子发挥作用。由此产生的 长丝是规则的，并且有大量的供应。通过分析 这些细丝的结构和起源，它们将解决重要的 有关tau丝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 自缔合和聚合？由此产生的 结构-活性-关系指向已知信号的参与 纤维生物发生中的转导机制？它的作用是什么 对tau自组装和细胞内稳定性的磷酸化？四个目标 都是为了解决这些问题。1)牛磺酸长丝的生产工艺 编队将被量化。使用一种新的组装测试，他们将 量化tau纤维形成的速率、程度和配体依赖性 在试管中。2)纤丝形成所需的tau的结构特征 将会被确定。这些研究将澄清结构要求 Tau聚合和潜在的配体结合部位的位置 用于开发治疗和治疗的高亲和力试剂 诊断应用程序。3)后翻译的影响 将评估对tau聚合动力学的修正。这些研究 将阐明磷酸化和糖基化对tau细丝的影响 队形。4)聚合反应对tau结构的影响 下定决心。使用圆二色光谱组合旋光法 和水动力测量，将确定tau是否采用 在长丝形成时定义的结构。结果将具有重要的意义 开发治疗药物的可行性的含义 预防和能够检测到的死前诊断试剂 纤维状病理的形成。

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.