NO production and S-nitrosylation in liver vasculature

肝血管系统中 NO 的产生和 S-亚硝基化

基本信息

批准号：
7431778
负责人：
YASUKO IWAKIRI
金额：
$ 13.68万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-15 至 2010-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The candidate has a three-year post-doctoral experience and is currently being proposed for an Associate Research Scientist appointment at the School of Medicine of Yale University. This application for a mentored K01 award is submitted with the goal of providing the candidate with the further training experience to function as an independent investigator working in the field of molecular biology and biochemistry of the liver. The project's long-term objective is to advance our understanding of nitric oxide (NO) signaling in liver physiology and pathophysiology. This is important for developing novel strategies to treat complications associated with liver diseases, since abnormal NO production by endothelial NO synthase (eNOS) plays a key role in vascular abnormalities observed in portal hypertension in liver cirrhosis. S-nitrosylation of proteins is an emerging area of investigation for NO-mediated physiological responses. We propose to investigate the mechanisms of S-nitrosylation, focusing on subcellular localization of eNOS and NO production, and identify the profiles of S-nitrosylated proteins in endothelial cells and liver cells using a proteomic approach. Our preliminary results suggest that there is a high concentration of NO gas at the eNOS localized area on the Golgi. This leads us to hypothesize that a gradient of NO formed in proximity to eNOS on the Golgi may be favorable to S-nitrosylation of proteins. The candidate proposes to 1) Elucidate if eNOS generates a gradient of NO in living cells, 2) Identify S-nitrosylated proteins in/on the Golgi complex using a proteomic approach, and 3) Examine protein S-nitrosylation in sinusoidal endothelial cells and hepatocytes. The Section of Digestive Diseases and the Department of Pharmacology as well as the Department of Cell Biology at Yale University are ideal for carrying out these studies because of the quality of their faculty, their experience as mentors, and the core facility at the Yale Liver Center. The School of Medicine has pledged protected time for the candidate during this further training period prior to functioning as an independent investigator.

描述（由申请人提供）：该候选人拥有三年的博士后经验，目前正在耶鲁大学医学院的副研究科学家任命。提出了这项指导K01奖的申请，目的是为候选人提供进一步的培训经验，以作为在分子生物学和肝脏生物化学领域工作的独立研究者的工作。该项目的长期目标是促进我们对肝生理和病理生理学中一氧化氮（NO）信号传导的理解。这对于制定与肝病相关的并发症的新型策略很重要，因为内皮NO合酶（ENOS）异常NO产生异常在肝肝硬化中观察到的血管异常中起关键作用。蛋白质的S-亚硝基化是无介导的生理反应的新兴领域。我们建议研究S-亚硝基化的机制，重点是eNOS和无生产的亚细胞定位，并使用蛋白质组学方法鉴定内皮细胞和肝细胞中S-亚硝基化蛋白的谱。我们的初步结果表明，高尔基体上的eNOS局部区域没有高浓度的气体。这使我们假设在高尔基体上与eNOS相邻的NO梯度可能有利于蛋白质的硝基化。候选人提议1）阐明eNOS是否在活细胞中产生NO的梯度，2）使用蛋白质组学方法鉴定高尔基体复合物中/上的S-硝基基化蛋白，3）3）检查正弦内皮细胞和肝细胞中蛋白S-硝基化的蛋白S-硝基化。耶鲁大学的消化系统疾病，药理学系以及耶鲁大学细胞生物学系非常适合进行这些研究，因为他们的教师的质量，作为导师的经验以及耶鲁大学肝脏中心的核心设施。在此进一步的培训期间，医学院已承诺为候选人提供保护时间，然后作为独立研究员的工作。