Peroxiredoxin 6 as an anti-oxidant enzyme

过氧化还原蛋白 6 作为抗氧化酶

• 批准号: 7861872
• 负责人: Aron B. FISHER
• 金额: $ 69.7万
• 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 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7861872
• 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; Hydrolysis; 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; Selenium; 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; 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. PUBLIC HEALTH RELEVANCE: We have characterized a novel enzyme which has two important activities that serve to protect the lungs against oxidant stress and also regulate metabolism of the lung surfactant. Our goals for the program are to evaluate the two activities of the enzyme to determine their relative importance in antioxidant defense, to investigate how the structure of the protein influences activity, and to determine methods for increasing the expression of the protein in lung cells. Understanding this role of peroxiredoxin 6 will facilitate this as a new target for increasing the ability of the lung cells to survive oxidant stress.

描述（由申请人提供）：该项目将评估一种新型的谷胱甘肽过氧化物酶的特性，该酶在肺中大大富集，并在肺抗氧化剂防御和肺表面活性剂代谢中起重要作用。最近描述的蛋白质称为过氧蛋白6（PRDX6或1-CYS过氧蛋白）代表唯一具有相对较高特异性活性的非亚度谷胱甘肽过氧化物酶。与经典的谷胱甘肽过氧化物酶不同，该酶的活性光谱中的一个重要特征是减少磷脂氢过氧化物的能力，例如在氧化剂胁迫期间膜磷脂的过氧化。 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蛋白质的磷脂酶（PLA2）活性。我们现在正在寻求5年的支持以扩展这些研究。具体目标1将利用PRDX6无效细胞中的“敲入”技术来评估两种活动（PRX，PLA2）对抗氧化剂保护的各自贡献。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）在抗氧化剂保护中的相对作用。具体目标2将通过KGF和地塞米松治疗研究PRDX6的诱导，我们的初步数据表明，这对PRDX6表达有协同作用。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，这是对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将利用细胞系统来分析PGST在PRDX6活性激活中的作用。拟议的研究将提供协调的努力，以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生化调节的新信息。这些信息可能会导致新的方法来提高肺耐受氧化应激的能力。 公共卫生相关性：我们已经描述了一种新型酶，该酶具有两种重要的活动，可保护肺部免受氧化剂胁迫，并调节肺表面活性剂的代谢。该程序的目标是评估酶的两种活性，以确定它们在抗氧化剂防御中的相对重要性，以研究蛋白质的结构如何影响活性，并确定增加肺部细胞中蛋白质表达的方法。了解过氧蛋白6的这一作用将有助于这是增加肺细胞在氧化剂胁迫下生存能力的新目标。