p47 binding partners in endothelial cell function

描述（由申请人提供）：缺血性和炎症性疾病 靶向血管内皮，并以增加的 细胞因子、强烈的氧化应激和增加的血管通透性。的 血管的这三个特征之间的关系的确切性质 然而，疾病定义不清，阻碍了治疗这些常见疾病的努力。 综合征 氧化剂只是最近才被认为是参与生理学过程。 信号传导过程，特别是由TNF-α启动的信号传导过程， 这些氧化剂的来源和目标是未知的。在简单的体外系统中， 氧化剂的易逝性质使得它们的作用具有高度的位点特异性， 使得它们的起始位点通常决定了它们的分子靶标。 最近的研究还表明，信号特异性，特别是在 MAP激酶系统也依赖于信号元件的适当共定位。 我们的初步数据表明， TNF-α诱导c-Jun氨基末端激酶（JNK）（一种MAP激酶）的活化 其活性形式与细胞骨架有关，在人内皮细胞中 细胞我们克隆了一个重要的信号接收衔接蛋白p47 phox 对于氧化酶，从人类内皮细胞库，并发现， p47 phox蛋白也定位于细胞骨架。此外，我们还 并筛选了HUVEC GAL 4文库中p47 phox的结合伴侣， 恢复了几个重要的基因，详细说明在身体的赠款。 我们假设，炎症细胞因子和生长因子的炎症信号传导可能与炎症反应有关。 血管细胞涉及吞噬细胞的氧化剂的位点特异性产生 氧化酶样复合物，其导致JNK和NF-κ B的下游活化。我们 进一步提出，氧化酶是拴在细胞色素相关的 通过与信号蛋白的特异性相互作用。 我们的总体目标是研究p47 phox的性质。 与三个潜在的结合靶点的相互作用，并探索 这些相互作用的功能意义，通过检查上游和 下游信号事件。通过更详细的了解 由于尼古丁诱导的促炎事件， 设计针对缺血性和炎症性血管的特定干预措施 疾病

DESCRIPTION (provided by the applicant): Ischemic and inflammatory diseases target the vascular endothelium and are marked by increased levels of cytokines, intense oxidative stress, and increased vascular permeability. The precise nature of the relationship between these three features of vascular diseases is ill defined, however, impeding efforts to treat these common syndromes. Oxidants have only recently been appreciated as participating in physiologic signaling processes, particularly those initiated by TNF-alpha, although the source and targets of such oxidants are not known. In simple in vitro systems, the evanescent nature of oxidants renders their effects highly site-specific, such that their site of origin generally dictates their molecular target. Recent studies have also revealed that signal specificity, particularly in the MAP Kinase system, also depends on proper colocalization of signaling elements. Our preliminary data implicate a phagocyte oxidase-like complex in the TNF-alpha induced activation of c-Jun amino terminal kinase (JNK), a MAP kinase whose active form is associated with the cytoskeleton, in human endothelial cells. We have cloned p47phox, an important signal-receiving adapter protein for the oxidase, from a human endothelial cell library, and found that the p47phox protein is also localized to the cytoskeleton. Further, we have made and screened a HUVEC GAL4 library for binding partners of p47phox, and have recovered several important genes detailed in the body of the grant. We hypothesize that inflammatory signaling by cytokines and growth factors in vascular cells involves site-specific production of oxidants by a phagocyte oxidase-like complex, which leads to downstream activation of JNK and NF-KB. We further propose that the oxidase is tethered to cytoskeleton-associated elements through specific interactions with signaling proteins. Our overall objectives will be to investigate the nature of p47phox interactions with three potential binding targets, and to explore the functional significance of these interactions by examining upstream and downstream signaling events. Through a more detailed understanding of cytokine-induced proinflammatory events, more opportunities will arise for designing specific interventions against ischemic and inflammatory vascular diseases.