Structural Study of Tau and Binding Partners

细胞骨架的基础结构是细胞生命的基本过程，如：运动，分裂，形态和细胞内运输。细胞骨架的微管（MT）成分是一种固有的动态结构，但MT在某些个体的神经系统中稳定超过100年。稳定的MT既维持突触连接，又充当突触功能所需的细胞内运输的高速公路。这些过程对衰老的影响很敏感，并且随着时间的推移与功能性突触的丧失相关。此外，细胞骨架的改变是神经退行性疾病如阿尔茨海默病和帕金森病的特征，其通常在生命后期表现。这项研究的目的是获得一些第一个结构特征的蛋白质参与稳定和运输的MT公路。tau蛋白被神经元用来稳定其MT，也是与阿尔茨海默病相关的两种主要蛋白质沉积物之一。历史上，由于其构象灵活性，tau的结构信息一直缺乏。这里的方法是解决tau与mapmodulin结合的结构，这将把tau锁定为单一构象，并规避微管蛋白的聚集趋势。结构信息将补充其他研究，并提供一个连贯的结构框架，用于理解不同构象状态的tau和微管之间的相互作用，为药物设计和预防或纠正tau沉积提供新的场所。目前还没有关于MT和细胞内转运之间相互作用的结构信息。我建议启动几个额外的MT结合域的研究，以澄清在分子水平上的微管成核，生长，减少，切断，稳定和调节，所需的细胞内运输的相互作用。

The cytoskeletal infrastructure underlies fundamental processes of cellular life such as: motility, division, morphology, and intracellular transport. The microtubule (MT) component of the cytoskeleton is an inherently dynamic structure and yet the MTs are stabilized for more than 100 years within the nervous system of some individuals. The stabilized MTs both maintain synaptic connections and serve as a highway for the intracellular transport required for synapsis function. These processes are sensitive to the effects of aging and correlated with the loss of functional synapsis over time. Additionally, alterations of the cytoskeleton are characteristic to neurodegenerative diseases such as Alzheimer's and Parkinson's disease, which typically manifest later in life. The research in this proposal is aimed at obtaining some of the first structural characterization of the proteins involved in stabilizing and trafficking upon the MT highway. The protein tau is used by neurons to stabilize its MTs and is also one of two major protein deposits associated with Alzheimer's disease. Historically, structural information on tau has be lacking due to its conformational flexibility. The approach here is to solve the structure of tau bound to mapmodulin, which will lock tau into a single conformation and circumvent the aggregation tendencies of tubulin. The structural information will complement other studies and provide a coherent structural framework for understanding the interactions between the different conformational states of tau and microtubules providing new venues for drug design and the prevention or correction of tau deposition. Currently there is an absence of structural information regarding interactions between MTs and intracellular transport. I propose to initiate studies on several additional MT binding domains in order to clarify at the molecular level the interactions involved in microtubule nucleation, growth, reduction, severing, stabilization, and regulation, that are required for intracellular transport.