CHARACTERIZATION GAMMA-SEC ACTIVITY CELL FREE SYSTEM

描述（来自申请人的摘要）：淀粉样蛋白的神经元产生 淀粉样前体蛋白（APP）的β（beta）似乎是一种必需的 在阿尔茨海默病（AD）的发展过程中是一个恒定的步骤。三个基因 与早发性AD有关这些基因是早老素1（PS1）， 早老素2（PS2）和APP本身。这些基因中的每一个突变都会导致 APP向聚集的Abeta种类的蛋白水解加工增加 更容易形成淀粉样斑块。三个独立的基因 暗示Abeta是阿尔茨海默病的致病因子， 治疗干预的重要目标。一种治疗方法是 AD是阻断产生Abeta的蛋白酶。这些蛋白酶还没有被 但产生Abeta的活动被称为beta和gamma 分泌酶γ分泌酶的作用是特别重要的， AD的进展，因为这种活动可以产生或多或少的聚集 Abeta的形式。本研究的目的是描述 的PS1 γ分泌酶活性，确定的亚细胞定位， γ分泌酶活性，最后鉴定γ分泌酶。这 将通过细胞提取物的生化操作来完成， 在体外测定中监测γ分泌酶活性。的识别 参与在γ分泌酶位点产生Abeta的蛋白酶将 进一步深入了解AD的分子机制， 增加了产生治疗抑制剂的机会， AD的进展。

DESCRIPTION (From the applicant's abstract): The neuronal production of amyloid beta (beta) from the amyloid precursor protein (APP) appears to be a necessary and invarieant step in the progression of Alzheimer's disease (AD). Three genes have been linked to early-onset AD. These genes are presenilin 1 (PS1), presenilin 2 (PS2), and APP itself. Mutations in each of these genes lead to the elevated proteolytic processing of APP into Abeta species that aggregate more readily and form amyloid plaques. The fact that three separate genes implicate Abeta as the causative agent of Alzheimer's disease makes this an important target for theraputic intervention. One way to create therapies for AD is to block the proteases that generate Abeta. These proteases have not been unidentified but the activities that create Abeta are termed beta and gamma secretase. The role of gamma secretase is especially important in the progression of AD since this activity can produce more or less aggregating forms of Abeta. It is the aim of this proposed study to characterize the role of PS1 in gamma secretase activity, identifying the subcellular localization of gamma secretase activity, and lastly to identity the gamma secretases. This will be accomplished through biochemical manipulation of cell extracts and monitoring gamma secretase activity in an in vitro assay. The identification of the protease(s) involved in producing abeta at the gamma secretase site will drastically further knowledge of the molecular mechanisms underlying AD and improve chances of creating theraputic inhibitors that slow or stop the progression of AD.