Peroxiredoxin 6 as an anti-oxidant enzyme

过氧化还原蛋白 6 作为抗氧化酶

• 批准号: 8445277
• 负责人: Aron B. FISHER
• 金额: $ 63.72万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8445277
• 关键词: Active Sites; Acute Lung Injury; Agonist; Alveolar; Antibodies; Antioxidants; Back; Binding; Biochemical; C-terminal; Cell membrane; Cell model; Cells; Characteristics; Complex; Cysteine; Cytoplasmic Protein; Cytosolic Phospholipase A2; DNA Sequence Rearrangement; Data; Dexamethasone; Docking; Elements; Endothelial Cells; Enzyme Activation; Enzymes; Epithelial; Epithelial Cells; Epithelium; Fatty Acids; Genes; Glucocorticoids; Glutathione Disulfide; Goals; Grant; Hydrogen Peroxide; Hyperoxia; In Vitro; Injury; International; Knock-in Mouse; Knock-out; Knockout Mice; Laboratories; Lead; Lipid Binding; Lipid Peroxidation; Lipid Peroxides; Lipids; Liposomes; Lung; Lysophospholipids; Mediating; Membrane; Membrane Lipids; Metabolism; Methods; Modeling; Molecular; Molecular Conformation; Mus; Mutate; Mutation; NADPH Oxidase; Natural regeneration; Nonesterified Fatty Acids; Null Lymphocytes; One-Step dentin bonding system; Oxidants; Oxidative Stress; Paraquat; Pathway interactions; Peroxidases; Peroxides; Phospholipase; Phospholipase A2; Phospholipids; Phosphorylation; Physiological; Play; Positioning Attribute; Post-Translational Protein Processing; Prevention; Progress Reports; Property; Protein Array; Protein Truncation; Proteins; Publications; Publishing; Pulmonary Surfactants; Reaction; Reagent; Reapplication; Recombinant Proteins; Reduced Glutathione; Regulation; Relative (related person); Research; Respiratory physiology; Response Elements; Role; Seminal; Site; Structure; Surface; Systems Analysis; Technology; Transferase; Transgenic Organisms; alveolar lamellar body; ascorbate; base; cell type; enzyme activity; enzyme pathway; glutathione peroxidase; hydroxy fatty acid; keratinocyte growth factor; knock-down; lung injury; mutant; novel; novel strategies; overexpression; oxidant stress; oxidation; peroxidation; peroxiredoxin; peroxiredoxin I; programs; promoter; protein expression; protein structure; public health relevance; repaired; response; surfactant; tert-Butylhydroperoxide; transcription factor

DESCRIPTION (provided by applicant): This project will evaluate the properties of a novel glutathione peroxidase enzyme that is considerably enriched in lungs and plays an important role in lung antioxidant defense and lung surfactant metabolism. This recently described protein called peroxiredoxin 6 (Prdx6 or 1-cys Peroxiredoxin) represents the only non-selenium glutathione peroxidase of relatively high specific activity. An important characteristic in the activity spectrum for this enzyme is its ability, unlike classical glutathione peroxidase, to reduce phospholipid hydroperoxides as, for example, peroxidation of membrane phospholipids during oxidant stress. During the past 4 years of support as a component project of a P-01 grant, we have demonstrated that Prdx6 plays a seminal role in defense of lungs against oxidant stress (hyperoxia, paraquat), that the promoter of the gene contains an antioxidant response element that is sensitive to the transcription factor Nrf2, and have developed a model based on lipid binding studies for the coordination of the peroxidase (Prx) and phospholipase (PLA2) activities of the protein. We are now seeking 5 years of support to extend these studies. Specific Aim 1 will utilize "knock-in" technology in the Prdx6 null cells to evaluate the respective contributions of the 2 activities (Prx, PLA2) to antioxidant protection. This aim will also directly compare the relative roles of Prdx6 and the other major glutathione peroxidase (GPx1) in antioxidant protection. Specific Aim 2 will study induction of Prdx6 by combined KGF and dexamethasone treatment, which our preliminary data indicate have a synergistic effect on Prdx6 expression. Specific Aim 3 will continue studies to evaluate structure-function characteristics of the protein with a special focus on phospholipid binding as a requirement for the phospholipid hydroperoxide peroxidase activity and the role of Prdx6 post-translational modifications. Specific Aim 4 will utilize cellular systems for analysis of the role of pGST in activation of Prdx6 activity. The proposed studies will provide a coordinated effort to investigate the role of this novel antioxidant enzyme in lung defense against oxidant stress and will provide new information concerning the biochemical regulation of its enzymatic activity. This information could lead to new approaches to increasing the ability of the lung to tolerate oxidant stress.

描述（由申请人提供）：该项目将评估一种新型谷胱甘肽过氧化物酶的特性，该酶在肺部大量富集，在肺抗氧化防御和肺表面活性剂代谢中发挥重要作用。最近描述的这种称为过氧化还原蛋白 6（Prdx6 或 1-cys 过氧化还原蛋白）的蛋白质代表了唯一具有相对较高比活性的非硒谷胱甘肽过氧化物酶。与经典的谷胱甘肽过氧化物酶不同，该酶的活性谱的一个重要特征是其减少磷脂氢过氧化物的能力，例如在氧化应激期间膜磷脂的过氧化。在过去 4 年作为 P-01 资助项目的一个组成部分的支持期间，我们证明了 Prdx6 在保护肺部抵御氧化应激（高氧、百草枯）方面发挥着重要作用，该基因的启动子含有对转录因子 Nrf2 敏感的抗氧化反应元件，并开发了一个基于脂质结合研究的模型，用于协调过氧化物酶 (Prx) 和 蛋白质的磷脂酶 (PLA2) 活性。我们现在正在寻求 5 年的支持来扩展这些研究。具体目标 1 将在 Prdx6 无效细胞中利用“敲入”技术来评估 2 种活性（Prx、PLA2）各自对抗氧化保护的贡献。该目标还将直接比较 Prdx6 和其他主要谷胱甘肽过氧化物酶 (GPx1) 在抗氧化保护中的相对作用。具体目标 2 将研究通过 KGF 和地塞米松联合治疗诱导 Prdx6，我们的初步数据表明这对 Prdx6 表达具有协同作用。具体目标 3 将继续研究以评估蛋白质的结构功能特征，特别关注磷脂结合作为磷脂氢过氧化物过氧化物酶活性的要求以及 Prdx6 翻译后修饰的作用。具体目标 4 将利用细胞系统分析 pGST 在激活 Prdx6 活性中的作用。拟议的研究将共同​​努力研究这种新型抗氧化酶在肺防御氧化应激中的作用，并将提供有关其酶活性的生化调节的新信息。这些信息可能会带来新的方法来提高肺部耐受氧化应激的能力。