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: 6372116
• 负责人: Mark E Fortini
• 金额: $ 31.25万
• 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/6372116
• 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（来自申请人的摘要）：本研究中提出的研究 申请旨在继续申请人的遗传和分子研究 早老素功能及其在细胞内运输和运输中的作用 其底物蛋白的蛋白水解加工。突变型人类早老素 影响淀粉样前体蛋白（APP）的蛋白水解，导致 阿尔茨海默病期间神经毒性淀粉样肽的加速积累 疾病。在模式生物秀丽隐杆线虫和果蝇中，早老素是 Notch/Lin-12 发育信号传导所需。早老素最近 已被证明可调节 Notch 受体期间的蛋白水解加工事件 成熟和信号传导可能类似于早老素依赖性 阿尔茨海默病中 APP 的裂解。最后，早老素也被 与哺乳动物细胞对凋亡刺激的细胞反应有关 细胞和果蝇。 将使用新分离的早老素在体内进行马赛克组织研究 基因突变体。初步实验揭示了整合素样表型 突变组织克隆，促使申请人分析其作用 使用遗传和生化方法研究早老素在整合素裂解中的作用 之前已被用来证明早老素对 缺口处理。这些研究可能揭示了早老素的共同特征 底物并导致更好地了解特定途径 蛋白质加工由早老素控制。该提案的中心目标是 开发大量新分子探针来剖析Notch 以比目前更高的分辨率进行处理。这些 试剂，包括新的抗体和表位标记的构建体，可以 区分Notch裂解产物，将与突变结合 和蛋白水解抑制研究以确定 Notch 的生化步骤 涉及早老素的加工。遗传和分子筛选 还将利用早老素相互作用因子进行 申请人最近发现 Presenilin 的保守 C 末端是 关键功能域。最后，详细的平行研究 Notch 和其他蛋白质的贩运将在缺乏的组织中进行 早老素或另一种具有已知亚细胞作用的蛋白质 运输，SERCA 型钙-ATP 酶。这些实验成为可能 通过申请人最近分离的钙-ATP酶突变体，他们将 解决蛋白质是否需要早老素这一尚未解决的问题 贩运或仅蛋白水解。这里提出的研究将澄清 Presenilin在Notch、APP等加工中的生化活性 蛋白质，并可能最终增加我们对分子原因的理解 阿尔茨海默病。