Peroxidation Profiles and Antioxidants

过氧化谱和抗氧化剂

• 批准号: 8375461
• 负责人: Ned Allen Porter
• 金额: $ 31.14万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-12 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8375461
• 关键词: Aerobic; Affinity; Alkynes; Amino Acid Sequence; Animals; Antioxidants; Arachidonic Acids; Azides; Biological; Biological Markers; Biological Models; Carbon Tetrachloride; Cells; Chemicals; Cholesterol Esters; Collaborations; Complex; Complex Mixtures; Coupled; Diet; Dietary intake; Disease; Docosahexaenoate; Drug toxicity; Environment; Equilibrium; Esters; Fatty Acids; Fish Oils; Fishes; Free Radicals; Glycerol; Glycerophosphates; Head; Health Benefit; High Pressure Liquid Chromatography; Human; Human Resources; Individual; Injury; Life; Link; Linoleic Acids; Lipid Peroxidation; Lipids; Liquid substance; Membrane; Membrane Lipids; Methodology; Methods; Molecular; Nature; Nucleic Acids; Organism; Oxidative Stress; Oxygen; Ozone; Pathway interactions; Peroxides; Phospholipids; Polyunsaturated Fatty Acids; Positioning Attribute; Process; Protein Chemistry; Proteins; Proteomics; Protocols documentation; Reaction; Reagent; Reducing Agents; Research; Residual state; Rodent; Screening procedure; Signal Transduction; Stress; Stress-Induced Protein; Tissues; Toxic effect; Unsaturated Fats; Vitamin E; adduct; analog; arachidonate; base; cycloaddition; environmental agent; environmental chemical exposure; fatty acid oxidation; free radical oxygen; human disease; hydroxypyridine; in vivo; interest; membrane model; novel; oxidation; peroxidation; polyunsaturated fat; programs; research study; response; tool

Peroxidation of polyunsaturated fatty acids (PUFAs) and their biologically relevant phospholipid esters is a complex reaction giving scores of possible products from a single molecular species. This process is a hallmark of diverse environmental chemical exposures, drug toxicities and oxidative stresses. In addition to the many peroxide products that form from polyunsaturated lipids, a set of reactive electrophiles is also generated. These electrophilic residuals of lipid peroxidation modify nucleic acids and proteins and in this way, the consequence of lipid oxidative degradation is distributed to other important biomolecules. This proposal outlines experiments that probe the chemical mechanisms of lipid peroxidation, provides a framework for understanding the oxidation of highly unsaturated lipids present in fish oils, describes novel new affinity-tags useful in the isolation of lipid electrophile-protein adducts and examines new phenolic antioxidants more potent than vitamin E. Highly unsaturated co-3 PUFAs are better reducing agents than more saturated co-6 lipids and we will look for consequences of this difference by analyzing peroxidation biomarkers formed in model membrane oxidations and in tissues and fluids of stressed animals on fish oil diets. Our affinity-tag lipids are analogs of natural lipids having a terminal alkyne substituted at the 00 position (co-yne) of fatty acid chains. This terminal alkyne undergoes "click" cycloaddition with biotinsubstituted azides, permitting "pull-down" of any proteins covalently attached to lipid-derived electrophiles bearing an (co-yne). The proposed research is based on the hypothesis that the chemical mechanisms of lipid peroxidation and the formation of electrophilic byproducts that are a hallmark of this process can be rationally defined. The affinity tags when coupled to powerful HPLC/MS/MS proteomics methods permit the structural identification of individual lipid-protein adducts even though such species are only a small part of a very complex mixture. Profiling of human THP-1 cells exposed to an oxidative stress will include studies in which affinity tag (co-yne) lipids are incorporated into the cells, permitting isolation of lipid-protein adducts. The electrophiles identified from phospholipids will form the basis of a screening program in collaboration with Projects 3 and 4 of the Program Project. New powerful pyridinol antioxidants will be studied in "proof of concept" in vivo rodent experiments.

多不饱和脂肪酸（PUFA）及其生物学相关的磷脂酯的过氧化是一种 复杂的反应从单个分子物种提供了数十种可能的产物。这个过程是 各种环境化学暴露，药物毒性和氧化应激的标志。此外 由多不饱和脂质形成的许多过氧化物产物，一组反应性电力也是 生成。这些脂质过氧化的亲电残留修饰核酸和蛋白质，在此中 方式，脂质氧化降解的结果分布在其他重要的生物分子上。这 提案概述了探测脂质过氧化化学机制的实验，提供了一种 了解鱼油中存在高度不饱和脂质的氧化的框架，描述了新型 新的亲和力标签可用于隔离脂质电力 - 蛋白质加合物并检查新酚类 抗氧化剂比维生素E更有效。 更多饱和的CO-6脂质，我们将通过分析过氧化来寻找这种差异的后果 在模型膜氧化以及鱼油上压力动物的组织和流体中形成的生物标志物 饮食。我们的亲和力标签脂质是碱替代末端的天然脂质的类似物 脂肪酸链的位置（二合一）。这个炔烃的末端经历了“单击” Cycloadition，用生物固定的构成 叠氮化物，允许在脂质衍生的电力的共同附着的任何蛋白质的“下拉” 带有（共同）。拟议的研究基于以下假设 脂质过氧化和这是该过程标志的亲电副产品的形成可以是 理性定义。当耦合到功能强大的HPLC/MS/MS蛋白质组学方法时，亲和力标签允许 即使这种物种只是一个的一小部分，即使单个脂质蛋白加合物的结构鉴定 非常复杂的混合物。暴露于氧化应激的人类THP-1细胞的分析将包括研究 将哪些亲和力标签（Co-Yne）脂质掺入细胞中，从而允许分离脂质 - 蛋白质加合物。 从磷脂中鉴定出的亲电器将构成合作中筛选程序的基础 随着计划项目的项目3和4。将研究新的强大吡啶醇抗氧化剂 概念“体内啮齿动物实验。