ACTIVITY OF THE ALZHEIMERS DISEASE PRESENIL IN PROTEIN

蛋白质中阿尔茨海默病早老性的活性

• 批准号: 6124218
• 负责人: Mark E Fortini
• 金额: $ 31.38万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6124218
• 关键词: Alzheimer's disease; Drosophilidae; amyloid proteins; developmental neurobiology; gene mutation; integrins; posttranslational modifications; presenilin; protein structure function; proteolysis

DESCRIPTION (From the applicant's abstract): Research proposed in this application seeks to continue the applicant's genetic and molecular studies on Presenilin function and its role in the intracellular trafficking and proteolytic processing of its substrate proteins. Mutant human Presenilins influence the proteolysis of amyloid precursor protein (APP), resulting in an accelerated accumulation of the neurotoxic amyloid peptides during Alzheimer's disease. In the model organisms Caenorhabditis and Drosophila, Presenilins are required for Notch/Lin-12 developmental signaling. Presenilins have recently been shown to regulate proteolytic processing events during Notch receptor maturation and signaling that may be analogous to the Presenilin-dependent cleavages of APP in Alzheimer's disease. Finally, Presenilins have also been implicated in the cellular response to apoptotic stimuli in both mammalian cells and Drosophila. Mosaic tissue studies will be performed in vivo using newly isolated Presenilin gene mutants. Preliminary experiments have revealed integrin-like phenotypes in the mutant tissue clones, prompting the applicants to analyze the role of Presenilin in integrin cleavage using the genetic and biochemical approaches that have been used previously to demonstrate the effects of Presenilin on Notch processing. These studies may reveal shared feature of Presenilin substrates and lead to a better understanding of the specific pathway of protein processing controlled by Presenilin. A central goal of this proposal is to develop an extensive collection of new molecular probes to dissect Notch processing at much higher resolution than is currently possible. These reagents, including new antibodies and epitope-tagged constructs that can discriminate among Notch cleavage products, will be combined with mutational and proteolysis inhibition studies to identify the biochemical steps of Notch processing that involve Presenilin. Genetic and molecular screens for Presenilin-interacting factors will also be performed, taking advantage of the applicant's recent finding that the conserved C-terminus of Presenilin is a crucial functional domain. Finally, detailed parallel studies on the trafficking of Notch and other proteins will be undertaken in tissues lacking either Presenilin or another protein with known effect of subcellular trafficking, the SERCA-type Calcium-ATPase. These experiments are made possible by the applicant's recent isolation of Calcium-ATPase mutants, and they will address the unresolved issue of whether Presenilin is required for protein trafficking or only proteolysis. The studies proposed here will clarify the biochemical activity of Presenilin in the processing of Notch, APP and other proteins, and may ultimately increase our understanding of the molecular causes of Alzheimer's disease.

描述（摘自申请人的摘要）：在此提出的研究 申请寻求继续申请人的遗传和分子研究 老年蛋白功能及其在细胞内贩运中的作用和 其底物蛋白的蛋白水解加工。突变的人类老年蛋白 影响淀粉样前体蛋白（APP）的蛋白水解，导致 阿尔茨海默氏症期间神经毒性淀粉样肽的加速积累 疾病。在模型生物体中，caenorhabditis和果蝇，老年蛋白是 Notch/Lin-12发育信号所需。 Presenilins最近有 已显示在Notch受体期间调节蛋白水解处理事件 可能类似于老龄素依赖性的成熟和信号传导 阿尔茨海默氏病中应用的分裂。最后，同龄人也一直在 与两个哺乳动物的细胞对凋亡刺激的反应有关 细胞和果蝇。 镶嵌组织研究将在体内使用新分离的老年蛋白进行体内进行 基因突变体。初步实验揭示了整联蛋白样表型 突变组织克隆，促使申请人分析 使用遗传和生化方法进行整联蛋白裂解中的老年蛋白 以前已用来证明Presenilin对 缺口处理。这些研究可能揭示了Presenilin的共同特征 底物并可以更好地理解 蛋白质加工由Presenilin控制。该提议的一个核心目标是 开发大量的新分子探针来剖析缺口 处理的分辨率比目前可能更高。这些 试剂，包括新抗体和表位标记的构建体 区分缺口裂解产品，将与突变结合 和蛋白水解抑制研究以鉴定Notch的生化步骤 涉及寄生素的处理。遗传和分子筛选 还将执行Presenilin的相互作用因素，利用 申请人最近的发现，即保守的Perenilin的C末端是一个 关键功能域。最后，详细的平行研究 将在缺乏的组织中进行槽口和其他蛋白质的运输 老年蛋白或其他蛋白质具有亚细胞的已知作用 贩运，Serca型钙ATPase。这些实验成为可能 通过申请人最近的钙-ATPase突变体的隔离，它们将 解决了是否需要蛋白质需要的尚未解决的问题 贩运或仅蛋白水解。这里提出的研究将阐明 Presenilin在Notch，App和其他的处理中的生化活性 蛋白质，最终可能会增加我们对分子原因的理解 阿尔茨海默氏病。