CORE--SCIENTIFIC

淀粉样前体蛋白(APP)的代谢命运受到调控 通过蛋白质磷酸化。蛋白激酶C(PKC)的激活，或 抑制蛋白磷酸酶-1(PP-1)刺激APP加工 通过非淀粉样变性的α-分泌酶途径。一个更伟大的 对控制细胞死亡的信号转导通路的理解 APP的细胞内贩运和处理，并管理 A-β淀粉样蛋白(A/β)的产生和分泌可能导致 为治疗阿尔茨海默病提供新的治疗靶点。 在该计划项目中提出了多学科的方法来 明确APP的细胞作用和APP的分子基础 依赖磷酸化的APP代谢调节。研究将会是 在组织复杂性的多个级别执行，包括 纯化分子的体外生化研究，细胞生物学 探索细胞内贩运事件的系统，以及 完整的动物分析APP代谢的变化 靶向缺失特定的PP-1亚型和两个内源性抑制物 PP-1 核心B旨在提供一系列技术支持服务，以 计划项目的其他成员。中央集权组织 支持功能将有助于实现可靠、高效且成本低廉的- 确保物资供应充足的有效手段。核心意志 负责充足物资的准备和维护 关键试剂(特定目标1)，包括纯化的蛋白激酶， 蛋白质磷酸酶以及这些酶的蛋白质和多肽抑制剂。 核心将提供建议并协调利用其他 蛋白质的化学支持功能，如多肽合成、蛋白质 测序和质谱学。APP的域特异性抗体 其代谢产物已成为APP研究的重要工具 正在处理。核心将保持目前使用的抗体库存， 并将产生额外的多克隆和磷酸化状态特异的 并协助开发其使用的分析方法 (具体目标2)。核心将维持基因敲除小鼠的群体 并提供建议研究的动物的所需数量 在项目3(具体目标3)中。这些将包括基因敲除小鼠 磷酸酶抑制物-1(已创建)、抑制物-2(待生产 在项目3中)、PP-1α和PP-1伽马(由其他 合作者)。L-1/L-2和PP-1阿尔法/伽马的“双”淘汰赛 是通过杂交繁殖产生的。

The metabolic fate of the amyloid precursor protein (APP) is regulated by protein phosphorylation. Activation of protein kinase C (PKC), or inhibition of protein phosphatase-1 (PP-1) stimulates processing of APP through the non-amyloidogenic, alpha-secretase pathway. A greater understanding of the signal transduction pathways that control the intracellular trafficking and processing of APP, and govern the production and secretion of the A-beta amyloid protein (A/beta), may lead to novel therapeutic targets for the treatment of Alzheimer's disease. A multi-disciplinary approach is proposed in this Program Project to define the cellular role of APP and the molecular basis of the phosphorylation-dependent regulation of APP metabolism. Studies will be performed at several levels of organizational complexity, encompassing in vitro biochemical studies with purified molecules, cell biological systems to explore intracellular trafficking events, and studies in intact animals analyzing changes in APP metabolism resulting from targeted deletion of specific PP-1 isoforms and two endogenous inhibitors of PP-1 Core B is designed to provide a range of technical support services to the other members of the Program Project. The centralized organization of support functions will facilitate a reliable, efficient, and cost- effective means to ensure an adequate supply of materials. The Core will be responsible for the preparation and maintenance of adequate supplies of key reagents (Specific Aim 1), including purified protein kinases, protein phosphatases and protein and peptide inhibitors of these enzymes. The Core will provide advice and coordinate the utilization of other protein chemical support functions, such as peptide synthesis, protein sequencing, and mass spectroscopy. Domain-specific antibodies for APP and its metabolites have been essential tools for the study of APP processing. The Core will maintain stocks of antibodies in current use, and will produce additional polyclonal and phosphorylation state-specific antibodies and assist in the development of assays for their use (Specific Aim 2). The Core will maintain the colonies of knock-out mice and provide the required number of animals that are proposed for study in Project 3 (Specific Aim 3). These will include the knock-out mice for phosphatase inhibitor-1 (already created), inhibitor-2 (to be produced in project 3), PP-1alpha, and PP-1gamma (to be provided by other collaborators). "Double" knock-outs of l-1/l-2 and PP-1alpha/gamma will be produced by cross-breeding.