PATHOGENESIS OF ALZHEIMERS DISEASE

这项拟议研究的长期目标是了解 B-淀粉样前体蛋白(APP)在 阿尔茨海默病的发病机制。最近的研究证实了 该APP是一种巡回膜蛋白，它展示了多个 细胞质分选信号。APP的细胞质尾部含有一个 与6-残基LDLR内化信号相关的序列GYENPTY FDNPVY以及符合4-残基的片段YTSI 基于酪氨酸的内化共识序列。这些信号是 由位于不同地点的分子分选机器识别 并确定细胞内的运输途径 应用程序就沿着这条路由。尽管有证据表明AB可以是 从细胞表面表达的APP产生，并且 需要内部化，目前还不清楚这是否是独家的 甚至是导致AB产生的主要贩运途径。 本申请的主要目标是确定如何 APP的贩运影响AB的生产以及APP是否 贩运受到其他基因产品的监管，这些基因产品与 阿尔茨海默病发病机制。为了实现这些目标，PI将采取 新出现的概念和方法进步的优势 膜蛋白分选领域，并将其应用于APP交易。 主要的实验方法将是开发转铁蛋白受体 (Tr)嵌合体，包括APP-tr嵌合体，所有定量生化 并对膜蛋白进行了形态研究。它的用途 受体嵌合体的特性允许这种受体配体的独特性质 用于定义在多种情况下的膜蛋白运输的系统 实验条件。发展起来的蛋白质修饰方法 通过研究tr嵌合体的贩运将被用来确定 修改后的应用程序交易如何影响AB的生产。

The long term goal of the proposed research is to understand the role of the B- amyloid precursor protein (APP) trafficking in the pathogenesis of Alzheimer's disease. Recent studies have established that APP is an itinerant membrane protein which displays multiple cytoplasmic sorting signals. The cytoplasm tail of APP contains a sequence GYENPTY related to the 6-residue LDLR internalization signal FDNPVY as well as the segment YTSI which conforms the 4-residue tyrosine-based internalization consensus sequence. These signals are recognized by molecular sorting machinery located at different sites within the cell and determine the intracellular trafficking pathways along which APP is routed. Although there is evidence that AB can be generated from APP expressed on the cell surface and that internalization is required, it is unclear whether this is the exclusive or even the major trafficking pathway leading to the production of AB. The primary objectives of the present application are to determine how trafficking of APP influences AB production and whether APP trafficking is regulated by other gene products implicated in the pathogenesis of AD. To accomplish these goals, the PI will take advantage of emerging conceptual and methodological advances in the field of membrane protein sorting and apply them to APP trafficking. The major experimental approach will be to exploit transferrin receptor (TR) chimers including APP-TR chimera that all quantitative biochemical and morphological studies of membrane proteins to performed. The use of the TR chimeras allows unique properties of this receptor-ligand system to be used to define membrane protein trafficking under a variety of experimental conditions. The methods developed to modify protein trafficking through the study of TR chimeras will be used to determine how altered APP trafficking affects AB production.