METABOLISM OF UNSATURATED AND HYDROXY FATTY ACIDS

不饱和脂肪酸和羟基脂肪酸的代谢

• 批准号: 2519274
• 负责人: HORST H SCHULZ
• 金额: $ 21.2万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519274
• 关键词: Escherichia coli; bioenergetics; cellular respiration; coenzyme A; enzyme mechanism; fatty acid metabolism; hydro lyase; hydroxy fatty acid; isomerase; laboratory rat; linolenate; microorganism metabolism; mitochondria; mutant; oleate; oxidation; oxidoreductase; peroxisome; respiratory enzyme; tissue /cell culture; transfection; unsaturated fatty acids

This project is designed to provide an in-depth understanding of the beta oxidation of unsaturated fatty adds, including polyunsaturated fatty adds. The focus of this study are the reactions and auxiliary enzymes which are specific to the beta-oxidation of unsaturated fatty acids. Most important is an investigation of the beta oxidation of unsaturated fatty adds with odd-numbered double bonds by the pathway involving delta 3,5, delta 2,4-dienoyl-CoA isomerase and 2,4-dienoyl CoA reductase. The contribution of this pathway to the mitochondrial beta-oxidation of 5- enoyl.CoA intermediates formed from linolenic add and oleic add will be determined. To facilitate such study with whole mitochondria, a mechanism-based inhibitor of delta 3,5, delta 2,4-dienoyl-CoA isomerase will be developed. The operation of this pathway in peroxisomes will be studied by determining the presence of delta 3,5, delta 2,4,-dienoyl-CoA isomerase in this organelle. The ongoing investigation of 2,4-dienoly-CoA reductase will be continued by cloning, sequencing and overexpressing the reductase gene from E. coli. The availability of a sufficent amount of the reductase will permit a study of its reaction mechanism. The substrate specificity of the peroxisomal reductase will be explored to aid in its purification. The notion that 3-hydroxyacyl-CoA epimerase may function in the metabolism of D-3-hydroxymyristic acid in E. coli will be investigated by isolating and characterizing epimerase mutants. The possible function of rat liver D-3-hydroxyacyl-CoA dehydratase, a peroxisomal enzyme essential for epimerase activity, in saturated fatty acid oxidation will be assessed by investigating its copurification with a suspected D-3-hydroxyacyl-CoA dehydrogenase. Finally the beta-oxidation of cis and trans unsaturated fatty acids in mitochondria will be compared with the aim of detecting and analyzing differences between their rates of degradation. This project will significantly increase the current knowledge of unsaturated fatty add beta-oxidation and thereby aid in the diagnosis of inherited diseases of fatty add oxidation and in assessing the metabolic consequences of trans fatty acid ingestion.

该项目旨在深入了解Beta 不饱和脂肪的氧化增加，包括多不饱和脂肪 添加。这项研究的重点是反应和辅助酶 特定于不饱和脂肪酸的β-氧化。最多 重要的是研究不饱和脂肪的β氧化 通过涉及三角洲3,5的途径添加奇数的双键， Delta 2,4-二烯酸-COA异构酶和2,4-二烯酰基COA还原酶。这 该途径对5--线粒体β-氧化的贡献 由Linolenic Add和Oleic添加形成的Enoyl.COA中间体将是 决定。为了促进整个线粒体的研究， 基于机理的抑制剂3,5，三角洲2,4-二烯酰基-COA异构酶 将开发。该途径在过氧化物酶体中的操作将是 通过确定三角洲3,5，三角洲2,4，-Dienoyl-COA的存在研究 该细胞器中的异构酶。正在进行的2,4-二烯coa的调查 还将通过克隆，测序和过表达还原酶继续 大肠杆菌还原酶基因。足够数量的可用性 还原酶将允许研究其反应机制。这 过氧化物酶体还原酶的底物特异性将被探索到 协助净化。 3-羟基酰基-COA分配酶可能的观念可能 在大肠杆菌中D-3-羟基甲酸的代谢中的功能 可以通过分离和表征分化酶突变体来研究。这 大鼠肝D-3-羟基乙酰辅酶A脱氢酶的可能功能，A 饱和脂肪中的分配酶活性必不可少的过氧化物酶体酶 酸氧化将通过研究其与 可疑的D-3-羟基酰基-COA脱氢酶。最后，β-氧化 将比较线粒体中的顺式和反式不饱和脂肪酸 目的是检测和分析其费率之间的差异 退化。该项目将大大增加电流 不饱和脂肪的知识增加了β-氧化，从而有助于 诊断脂肪的遗传疾病增加氧化并评估 反式脂肪酸摄入的代谢后果。