PHOSPHORYLATION AND SPATIAL LOCALIZATION OF TAU PROTEIN

TAU 蛋白的磷酸化和空间定位

• 批准号: 2546430
• 负责人: Gloria Lee
• 金额: $ 20.17万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2546430
• 关键词: PC12 cells; axon; biological signal transduction; cell differentiation; cell membrane; cytoskeleton; immunocytochemistry; immunofluorescence technique; microtubules; mutant; neurons; phosphorylation; tau proteins

Tau protein is a neuronal microtubule associated protein that promotes microtubule assembly and functions in axonal development. Additionally, abnormally phosphorylated tau is an integral component of neurofibrillary tangles and thus may contribute to Alzheimer's disease pathogenesis. While much is known about the structure and function of tau in microtubule assembly, the mechanisms that govern tau's spatial localization and that regulate tau phosphorylation, and thus its activity, are poorly understood. The applicant has determined that in transfected neuronal cells, the amino terminal of tau localizes to the cell membrane and creates a dominant negative effect on neurite outgrowth. She hypothesizes that tau mediates microtubule-membrane interactions that are required for neuritic development. Here she proposes to test this hypothesis. The molecular basis for tau's association with the membrane will be elucidated and the significance of this association in neuronal differentiation will be determined. To determine the molecular basis of tau's association with the membrane, the applicant will 1) identify the area of the amino terminus of tau that binds to the membrane, 2) determine the biochemical characteristics of the binding between tau and purified membrane, and 3) identify the component(s) of the membrane that interact with tau using co-immunoprecipitation. To investigate the role of membrane associated tau during neuronal differentiation, she will 1) examine membrane associated tau in non-transfected differentiated neuronal cells and in primary cultured neurons and 2) investigate the effect of over expressing the membrane associated tau domain during neuronal differentiation. These studies will provide new insights into the function of tau's amino terminus and into tau's role in the transduction of extracellular signals to the cytoskeleton.

Tau蛋白是一种神经元微管相关蛋白， 微管组装和功能在轴突发育。此外，本发明还 异常磷酸化的tau蛋白是神经胶质瘤的一个组成部分， 缠结，因此可能有助于阿尔茨海默病的发病机制。 虽然对微管中tau蛋白的结构和功能了解很多， 组装，控制tau蛋白空间定位的机制， 调节tau蛋白磷酸化，从而调节其活性， 明白申请人已经确定，在转染的神经元中， 在细胞中，tau的氨基末端定位于细胞膜， 对神经突生长产生了显著的负面影响。她假设 tau蛋白介导微管-膜相互作用， 神经炎发展在这里，她打算检验这个假设。 tau蛋白与细胞膜结合的分子基础是 阐明了这种关联在神经元中的意义， 差异化将被确定。来确定 tau与膜的结合，申请人将1）确定 与膜结合的tau的氨基末端的面积，2）确定 tau蛋白与纯化的 膜，以及3）识别相互作用的膜的组分 使用免疫共沉淀法与tau结合。调查的作用 在神经元分化期间，她将1） 在未转染的分化的神经元中检查膜相关的tau 细胞和原代培养的神经元，2）研究 在神经元生长过程中过度表达膜相关的tau结构域 分化 这些研究将为tau蛋白氨基的功能提供新的见解 末端并进入tau蛋白在细胞外信号转导中的作用 到细胞骨架。