Vesicular transport of the amyloid precursor protein

描述（由申请人提供）：阿尔茨海默病（AD）的主要特征之一是老年斑，其含有通过淀粉样前体蛋白（APP）加工形成的Ab。细胞内运输在APP加工中起着至关重要的作用，但参与控制APP细胞内交通的机制仍然没有得到充分的表征。两种蛋白质Mint和FE65在APP加工中发挥相反的作用：Mint抑制Ab的产生，FE65增加Ab的产生。最近的数据表明，薄荷糖可能作为囊泡外壳蛋白，因此薄荷糖可能通过控制APP流量来调节APP处理。这项建议将建立生化分析，将确定参与APP囊泡运输的分子组成。将使用膜募集测定来测试APP作为货物受体的假设，该货物受体以被FE 65阻止的ARF依赖性方式将薄荷募集到膜上。将进行体外出芽试验，以确定含APP囊泡的出芽是否需要ARF和薄荷。由于APP加工依赖于APP运输，这些数据将阐明APP靶向非致病途径的机制，从而确定AD的新治疗靶点。

DESCRIPTION (provided by applicant): One of the primary characteristics of Alzheimer's disease (AD) is senile plaques containing Ab which is formed by the processing of amyloid precursor protein (APP). Intracellular transport plays a crucial role in APP processing, but the mechanisms involved in controlling APP intracellular traffic remain insufficiently characterized. Two proteins, Mints and FE65 play opposite roles in APP processing: Mints inhibit and FE65 increases the production of Ab. Recent data suggest that Mints may function as a vesicular coat protein and thus Mints may regulate APP processing by controlling APP traffic. This proposal will establish biochemical assays that will define the molecular components involved APP vesicular trafficking. Membrane recruitment assays will be used to test the hypothesis that APP acts as a cargo receptor that recruits Mints to the membrane in an ARF-dependent manner that is prevented by FE65. In vitro budding assays will be performed to determine if the budding of APP-containing vesicles requires ARF and Mints. Because APP processing depends on APP trafficking, these data will elucidate the mechanisms by which APP is targeted to a nonpathogenic pathway and consequently identify novel therapeutic targets for AD.