Role of Protein Nitration In Altering The Rho/Rac1 Balance In Acute Lung Injury

蛋白质硝化在改变急性肺损伤中 Rho/Rac1 平衡中的作用

• 批准号: 8198062
• 负责人: Stephen M Black
• 金额: $ 36.22万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-10 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8198062
• 关键词: 2-tyrosine; 3-Dimensional; Acute Lung Injury; Attenuated; Binding Sites; Biochemical; Blood; Blood Vessels; Data; Development; Disease; Drops; Endothelial Cells; Endothelium; Enzymes; Equilibrium; Event; Family; Generations; Goals; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; In Vitro; Instruction; Investigation; Lung; Maintenance; Maps; Mass Spectrum Analysis; Mediating; Modification; Molecular; Molecular Conformation; Molecular Models; Mus; N,N-dimethylarginine; Nitrates; Nucleotides; Outcome; Permeability; Peroxonitrite; Pharmacotherapy; Phosphorylation; Physiological; Play; Property; Proteins; Publishing; Reactive Nitrogen Species; Reagent; Regulation; Research; Role; Secondary to; Signal Transduction; Surface; Surgical Flaps; Testing; Therapeutic Agents; Tissues; Tyrosine; Tyrosine Phosphorylation; Work; base; clinically relevant; dimethylargininase; human NOS3 protein; in vivo; lung injury; member; molecular dynamics; molecular modeling; mortality; mouse model; nitration; novel; novel therapeutics; prevent; repaired

PROJECT SUIVIMARY (See instructions): An important function of the endothelium lining the inner surface of blood vessels is to provide a selective barrier between blood and the surrounding tissues. During the development of acute lung injury (ALI) the endothelial barrier is weakened, leading to increased permeability. It is well known that the family of small ras homology (Rho) GTPases (RhoA, Rad, Cdc42) play a crucial role in the maintenance of endothelial barrier properties. The two best-characterized members of the Rho GTPases: Rho A and Rad, appear to regulate endothelial barrier function in an antagonistic manner. Thus, the activation of Rho A impairs barrier function whereas Rad appears to support barrier integrity. In addition, our recent studies have shown that during the development of ALI in the mouse lung the activities of RhoA and Rad are regulated in an opposing manner such that RhoA activity is increased and Rad activity is attenuated. Together these changes would favor barrier disruption. However, the mechanism by which this opposing regulation occurs unresolved and is the major focus of this project. We will evaluate the mechanisms by which uncoupled eNOS leads to modulation of RhoA/Rad balance through nitration-mediated modifications. We will also determine if preventing RhoA and Rad nitration is barrier protective in vitro and reduces lung injury in both G' and G^-mouse models of ALI, in vivo. It is anticipated that this Project using state-of-the-art cellular, molecular, biochemical, and physiological approaches that will not only increase our understanding of the mechanisms by which RhoA and Rad are regulated during both G(-)- and G(+) -induced ALI but will facilitate the development of new strategies and targets for the treatment of a disease that has not seen a significant drop in mortality in 40 years.

项目随附资料（见说明）： 内衬血管内表面的内皮的重要功能是在血液和周围组织之间提供选择性屏障。在急性肺损伤（ALI）的发展过程中，内皮屏障被削弱，导致通透性增加。众所周知，小ras同源性（Rho）GTP酶家族（RhoA、Rad、Cdc 42）在维持内皮屏障特性中起关键作用。Rho GTP酶的两个最佳表征成员：Rho A和Rad似乎以拮抗方式调节内皮屏障功能。因此，Rho A的激活损害屏障功能，而Rad似乎支持屏障完整性。此外，我们最近的研究表明，在小鼠肺中ALI的发展过程中，RhoA和Rad的活性以相反的方式调节，使得RhoA活性增加而Rad活性减弱。所有这些变化都有利于屏障的破坏。然而，这种相反的调节发生的机制尚未解决，这是本项目的主要焦点。我们将评估通过硝化介导的修饰，解偶联的eNOS导致RhoA/Rad平衡调节的机制。我们还将确定预防RhoA和Rad硝化是否在体外具有屏障保护作用，并在体内减少ALI的G'和G^-小鼠模型中的肺损伤。预计该项目使用最先进的细胞，分子，生物化学和生理学方法，不仅将增加我们对G（-）和G（+）诱导的ALI期间RhoA和Rad调节机制的理解，而且将促进新策略和靶点的开发，用于治疗40年来死亡率未显着下降的疾病。