ENDOTHELIAL CELL HYPOXIA-ASSOCIATED PROTEINS

在体内，生物体和它们的驻留组织和细胞经常被 暴露于环境氧气的减少。 因此，急性耐受性 缺氧以及在某些情况下对慢性缺氧的适应，对于 生存和不同细胞之间的差异。 一些适应性 已经提出了允许低氧耐受性发展的策略 其他的，都是未知数。 我们最近发现了一个 耐缺氧内皮细胞诱导特异性应激蛋白 细胞暴露于急性和慢性缺氧。 这些蛋白质， 称为缺氧相关蛋白（HAP），是由 环境缺氧，不同于其他描述的应激蛋白， 如热休克或葡萄糖调节蛋白， 在RNA水平上进行调节。 为了表征内皮细胞HAP， 为了更全面地规管长者住屋计划，我们会：1）研究长者住屋计划本地化的问题 肺动脉内皮细胞内; 2）研究 HAPs和其他应激蛋白的关系，作为保护免受 缺氧、热休克和葡萄糖剥夺的毒性作用; 通过直接蛋白质测序和从低氧细胞中克隆HAPs cDNA， 刺激的肺动脉内皮细胞cDNA文库; 4）使用 分离的蛋白或从HAPs cDNA推导的蛋白序列， 开发分子探针; 5）利用抗体和cDNA探针， 研究HAPs转录和翻译调控， 培养的细胞和缺氧组织，并确定 改变HAPs表达对内皮细胞缺氧反应的影响。 这些 研究将提供深入了解HAPs在生物学上的调节， 和遗传水平，并可提供细胞中缺氧的标志物， 组织中

In vivo, organisms and their resident tissues and cells are frequently exposed to decreases in ambient oxygen. Therefore, tolerance to acute hypoxia and, in some cases, adaptation to chronic hypoxia, is critical to survival and varies among different cells. Some of the adaptive strategies permitting development of hypoxia tolerance have been proposed and evaluated; others are likely unknown. We have recently identified a specific set of stress proteins induced in hypoxia tolerant endothelial cells during exposure to both acute and chronic hypoxia. These proteins, termed hypoxia associated proteins (HAPs), are specifically induced by environmental hypoxia, are distinct from other described stress proteins, such as heat shock or glucose-regulated proteins, and are likely regulated at the RNA level. To characterize endothelial cell HAPs and their regulation more fully, we will: 1) examine the localization of HAPs within pulmonary arterial endothelial cells; 2) investigate the relationship of HAPs and other stress proteins as protection against the toxic effects of hypoxia, heat shock, and glucose deprivation; 3) isolate HAPs by direct protein sequencing and by cloning HAPs cDNA from a hypoxia stimulated pulmonary arterial endothelial cell cDNA library; 4) use the isolated protein or the protein sequence deduced from HAPs cDNA to develop molecular probes; and 5) use antibody and cDNA probes to investigate transcriptional and translational regulation of HAPs in cultured cells and hypoxic tissues and to determine the effect of altering HAPs expression on endothelial cell response to hypoxia. These studies will provide insight into the regulation of HAPs at the biologic and genetic level and may provide markers for hypoxia in cells and tissues.