CONTROL OF PHAGOCYTE NADPH OXIDASE BY CYTOSOLIC PROTEINS

胞浆蛋白对吞噬细胞 NADPH 氧化酶的控制

• 批准号: 6475875
• 负责人: PAUL G HEYWORTH
• 金额: $ 29.17万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6475875
• 关键词: NAD(P)H dehydrogenase; actin binding protein; biological signal transduction; chronic granulomatous disease; clinical research; cytoplasm; enzyme activity; enzyme biosynthesis; enzyme inhibitors; fluorescence microscopy; guanine nucleotide exchange factors; guanosinetriphosphatase activating protein; guanosinetriphosphatases; human subject; immunofluorescence technique; inflammation; laboratory rabbit; neutrophil; phagocytosis; vesicle /vacuole

DESCRIPTION (Applicant's Description Verbatim): The superoxide(02-)-generating NADPH oxidase of phagocytes is a potent source of reactive oxygen species (ROS). Its importance to innate immunity is manifested by the inherited syndrome chronic granulomatous disease, in which oxidase activity is absent and patients are susceptible to life-threatening microbial infections. O2-and its derivatives also cause severe tissue damage, contributing to inflammatory diseases such as ischemia-reperfusion injury and acute respiratory distress syndrome, and their prolonged generation in chronic inflammation can lead to dysplasia and malignancy. ROS produced by homologous systems in non-phagocytes are implicated in abnormal cell growth. The long-term objectives of the proposed research are to understand the mechanisms that control NADPH oxidase assembly and activity, and to identify steps that can be inhibited to minimize tissue damage. These four specific aims address gaps in knowledge that impede achievement of these goals. (1) Identify the processes of molecular recognition that encode the complex of cytosolic oxidase proteins to associate with the flavocytochrome to form a isolated phagosomes and cellular fractions, will be used to study the formation of the active enzyme and determine the roles of coronin and cofilin in controlling its assembly. 2) Analyze interactions between NADPH oxidase components, and identify factors that promote their association and dissociation. A biosensor instrument will be used to quantitate interactions between oxidase-related proteins and study how they are modified by changes that mimic intracellular signals. Development of this technique will provide an assay for molecules designed to inhibit oxidase activation. 3) Identify and isolate GTPase activating proteins and guanine nucleotide exchange factors for Rac2 and study their involvement in controlling Rac and NADPH oxidase activity. The identities of the regulatory proteins that control Rae activity in the neutrophil by altering its GDP/GTP-bound state are not known. These proteins are likely to be important in the negative and positive regulation of 02- generation. 4) Identify signaling events and molecules that terminate superoxide generation. The processes that switch off the enzyme have important clinical implications for inflammatory disease, but they are not well understood. Cell-free oxidase systems and intact cells will be used to determine if a) phosphoinositides; b) changes in the phosphorylation state of phox proteins; and c) changes in the GDP/GTP-bound state of Rae are negative regulators of the enzyme.

描述（申请人的逐字描述）： 吞噬细胞的NADPH氧化酶是活性氧的有力来源 （ROS）。它对先天免疫的重要性表现在 综合征慢性肉芽肿性疾病，其中氧化酶活性缺失， 患者容易受到威胁生命的微生物感染。O2-及其 衍生物也会导致严重的组织损伤，导致炎症反应。 缺血-再灌注损伤和急性呼吸窘迫等疾病 综合征，以及它们在慢性炎症中的长期生成可导致 发育不良和恶性肿瘤。非吞噬细胞同源系统产生的活性氧 与异常细胞生长有关的长期目标 建议的研究是了解控制NADPH氧化酶的机制， 组装和活动，并确定可以抑制的步骤，以尽量减少 组织损伤这四个具体目标旨在解决阻碍实现这些目标的知识差距。 实现这些目标。(1)识别分子识别的过程 编码胞浆氧化酶蛋白复合物， 黄细胞色素形成一个孤立的吞噬体和细胞部分，将 用于研究活性酶的形成，并确定 coronin和cofilin控制其组装。2)分析互动 NADPH氧化酶组分之间的关系，并确定促进其 结合和解离。生物传感器仪器将用于定量 氧化酶相关蛋白质之间的相互作用，并研究它们如何被修饰 通过模仿细胞内信号的变化。这项技术的发展将 提供了设计用于抑制氧化酶活化的分子的测定。第三章 鉴定和分离GT3活化蛋白和鸟嘌呤核苷酸交换 因子Rac 2和研究他们参与控制Rac和NADPH 氧化酶活性控制Rae的调节蛋白的身份 通过改变其GDP/GTP-结合状态而在中性粒细胞中的活性是未知的。 这些蛋白质很可能是重要的消极和积极的 02代调控。4)识别信号事件和分子， 终止超氧化物生成。关闭酶的过程 重要的临床意义，炎症性疾病，但他们并不好 明白无细胞氧化酶系统和完整细胞将用于 确定是否a）磷酸肌醇; B）磷酸化状态的变化 phox蛋白;和c）Rae的GDP/GTP结合状态的变化是负的 酶的调节剂。