NEW MOLECULAR INTERACTOR FOR TAU PROTEIN

在衰老过程中，神经元细胞中的一些蛋白质结构发生变化， 组织中 研究得最充分的变化是发生在 与年龄相关的神经退行性疾病。 这一建议的重点是tau 蛋白质，一种神经元微管相关蛋白， 神经元缠结的组成部分，并在一个新发现的人类 与tau相互作用的蛋白质。 神经纤维缠结不正常 在几种神经退行性疾病中发现的神经元内细丝， 例如阿尔茨海默氏病，以及正常的老年大脑。 这些 缠结代表了tau蛋白结构在 衰老 因为缠结形成的细胞机制 尚未阐明，我们使用酵母双杂交系统来鉴定 与tau蛋白相互作用的蛋白质。 这些蛋白质可能在 在缠结形成之前或之后的病理事件中。 我们有 鉴定了一种含有WD重复序列的新的人类蛋白质， τ的 WD重复序列是仅在调节蛋白中发现的基序。 为了支持这种新的人类蛋白质的调节作用，数据库 研究表明，这种蛋白质的小鼠同源物存在， 在小鼠发育过程中形成模式至关重要。 我们 我认为这种新的蛋白质除了它的作用外还有其他的功能 在发展中，它将具有更多的重要职能， 正常成人大脑和阿尔茨海默病的神经病理学。 本提案的具体目标是：调查相互作用 tau和WD重复蛋白在体内和体外之间的关系，以及II. 人胎儿WD重复序列蛋白的空间定位， 中年人、老年人和患病的大脑。 在具体目标I中， 我们将使用哺乳动物双杂交系统来证实这种相互作用。 在细胞中。 我们将使用体外结合测定来1）定位 2）确定每个蛋白质的化学计量， 结合，和3）确定微管是否干扰 互动 我们还将确定WD重复蛋白是否可以 促进tau的聚合。 在具体目标II中，我们将 制备几种特异性抗WD重复序列蛋白的抗血清，并进行鉴定 并将内源性蛋白定位在培养的人类细胞中。 在 此外，人胎儿、中年成人、老年成人和AD脑 将探测组织切片以确定定位 WD重复蛋白相对于异常磷酸化的tau蛋白的表达， 平野体。 我们还将确定是否存在与年龄相关的变化 WD重复序列蛋白的结构和定位。 这项建议 试图阐明tau蛋白与其他蛋白相互作用的分子基础， 和WD重复序列蛋白，并提供对 这种新的人类蛋白质。

During aging, several protein structural changes take place in neuronal tissues. Among the most well studied changes are those which occur in age-related neurodegenerative diseases. This proposal focuses on tau protein, a neuronal microtubule associated protein that is a primary component of neurofibrillary tangles, and on a newly identified human protein that interacts with tau. Neurofibrillary tangles are abnormal intraneuronal filaments found in several neurodegenerative diseases, such as Alzheimer's disease, and also in normal aged brain. These tangles represent a dramatic change in tau protein structure during aging. Because the cellular mechanisms underlying tangle formation have yet to be elucidated, we used the yeast two hybrid system to identify proteins that interact with tau. Such proteins potentially have a role in pathological events preceding or following tangle formation. We have identified a new human protein containing WD repeats that interacts with tau. WD repeats are motifs exclusively found in regulatory proteins. In support of a regulatory role for this new human protein, database searching revealed that the mouse homolog of this protein exists and is critically required for pattern formation during mouse development. We propose that this new protein had additional functions beyond its role in development and that it will have additional important functions in normal adult brain and in the neuropathology of Alzheimer's disease. The Specific Aims of this proposal are I. Investigate the interaction between tau and the WD repeat protein in vivo and in vitro and II. Determine the spatial localization of WD repeat protein in human fetal, middle aged adult, aged adult, and diseased brain. In Specific Aim I, we will use the mammalian two hybrid system to confirm the interaction in cells. We will use in vitro binding assays to 1) map the site(s) of interaction on each protein, 2) determine the stoichiometry of the binding, and 3) determine if microtubules interfere with the interaction. We will also determine if the WD repeat protein can facilitate the polymerization of tau. In Specific Aim II, we will prepare several specific antiserum to the WD repeat protein and identify and localize the endogenous protein in cultured human cells. In addition, human fetal, middle aged adult, aged adult, and AD brain tissue sections will be probed in order to determine the localization of the WD repeat protein relative to abnormally phosphorylated tau and Hirano bodies. We will also determine if there are age related changes in the WD repeat protein structure and localization. This proposal seeks to elucidate the molecular basis for the interaction between tau and the WD repeat protein and to provide insights into the function of this new human protein during aging.