REGULATION OF NF-KAPPA B BY NITRIC OXIDE IN THE LUNG

肺中一氧化氮对 NF-KAPPA B 的调节

• 批准号: 6652525
• 负责人: HARVEY E MARSHALL
• 金额: $ 12.51万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6652525
• 关键词: cell line; lung; nitric oxide; nuclear factor kappa beta; protein kinase; recombinant proteins; respiratory epithelium; thiols

NF-Kappa B is a transcription factor which regulates numerous inflammatory response genes. Cellular production of nitric oxide (NO) is increased in such inflammatory states. Depending upon the cell type, stimulus, concentration, and form of delivery, NO has been found to be either stimulatory or inhibitory to NF-Kappa B. This proposed research is designed to test the effects of NO or NO-related molecules on NF-Kappa B activity in the respiratory epithelium. Our preliminary studies indicate that NO has an inhibitory effect on NF-Kappa B in respiratory epithelial cells. The mechanism(s) by which NO exerts this effect is not understood. We hypothesize that NO regulates NF-Kappa B via the formation of a nitrosothiol on the p50-p65 heterodimer or another protein in the activation pathway. To investigate this hypothesis we plan to: 1. Determine the effects of exogenous and endogenous NO or NO-related molecules on NF-Kappa B activity in respiratory epithelial cells and determine whether these effects are sensitive to intracellular thiol levels. 2. Analyze recombinant NF-Kappa B proteins in vitro to determine whether they can be nitrosylated and at which site(s) this occurs on the molecule. 3. Determine whether these S-nitrosylated NF-Kappa B proteins are present intracellularly. 4. Determine the mechanism by which S-nitrosylation of these proteins alters their cellular function. The applicants long term goals are to become an independent investigator studying the regulation of gene transcription in the lung, in particular, as it relates to oxidative and nitrosative stress. With this award, the applicant will obtain the necessary training and experience in molecular biological techniques to achieve this goal.

NF-KAPPA B是调节大量炎症反应基因的转录因子。 在这种炎症状态下，一氧化氮（NO）的细胞产生增加。 根据细胞类型，刺激，浓度和递送形式，NO被发现对NF-kappaB是刺激性或抑制性B。该提出的研究旨在测试NO或NO相关分子对NF-KAPPA B活性在呼吸性上皮中的影响。 我们的初步研究表明，NO对呼吸性上皮细胞中的NF-kappa B具有抑制作用。不理解不发挥这种作用的机制。 我们假设NO通过P50-P65异二聚体或激活途径中的另一种蛋白质在P50-P65异二聚体上形成NF-KAPPA B。 为了研究这一假设，我们计划：1。确定呼吸性上皮细胞中外源性和内源性或无关分子对NF-kappa B活性的影响，并确定这些作用是否对细胞内硫醇水平敏感。 2。在体外分析重组NF-kappa B蛋白，以确定它们是否可以被硝基基化以及在哪个位点发生在分子上。 3。确定这些S-亚硝基化的NF-kappa B蛋白是否存在细胞内。 4。确定这些蛋白质的S-亚硝基化改变其细胞功能的机制。申请人的长期目标是成为研究肺中基因转录调节的独立研究者，因为它与氧化和硝化应激有关。 通过此奖项，申请人将获得必要的分子生物学技术培训和经验，以实现这一目标。