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: 3341868
• 负责人: HORST H SCHULZ
• 金额: $ 13.09万
• 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 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3341868
• 关键词: Escherichia coli; acid thiol ligase; antibody; cis trans isomerization; eicosanoid metabolism; enzyme inhibitors; enzyme mechanism; fatty acid metabolism; heart metabolism; hydroxy fatty acid; isomerase; laboratory rat; leukotrienes; liver metabolism; messenger RNA; mitochondria; oxidation reduction reaction; oxidoreductase; peroxisome; somatotropin; unsaturated fatty acids

This study is designed to answer some unresolved questions about the metabolism of polyunsaturated fatty acids (PUFA) and of hydroxy fatty acids related to leukotrienes. In order to fully understand the function of the peroxisomal 3-hydroxyacyl-CoA epimerase in the degradation of PUFA, the rat liver enzyme will be purified and characterized. A specific inhibitor of 2,4-dienoyl-CoA reductase will be designed and an E. coli mutant lacking 2,4-dienoyl-CoA reductase will be isolated. Both the reductase inhibitor and the E. coli mutant will be used to determine the contribution of the old epimerase-dependent pathway to the degradation of PUFA in rat liver peroxisomes and E. coli. The inhibitor of 2,4-dienoyl-CoA reductase will also be used to determine, if the reaction catalyzed by the reductase is rate-limiting in the oxidaiton of PUFA. The presence or absence of cis-3-trans-2-enoyl-CoA isomerase in rat liver peroxisomes will be established and if present, the enzyme will be isolated and characterized. We will continue to study the hormonal regulation of 2,4-dienoyl-CoA reductase and of the oxidaiton of PUFA. This study necessitates the purification of mitochondrial 2,4-dienoyl-CoA reductase and the preparation of antireductase antibodies which will be used to determine the levels of reductase in mRNA. Additionally, differences between the rates of oxidation of cis and trans unsaturated fatty acids will be determined with rat heart mitochondria. The oxidation of hydroxy fatty acids including 5-hydroxy-6,8,11,13-eicosatetraenoic acid (5-HETE) in mitochondria and peroxisomes will be studied, as will be the inhibition of Beta-oxidation by these and some other potential inhibitory compounds. An assessment of the effectiveness of inhibitors of Beta-oxidation in liver and heart necessitates the purification and characterization of acyl-CoA synthetases present in the mitochondrial matrix. Finally, we will test 3-decynoic acid and other possible inhibitors of acyl-CoA oxidase for their effectiveness as inhibitors of peroxisomal fatty acid oxidation. If effective, such compounds will be used to evaluate the contribution of peroxisomal Beta-oxidation to the hepatic metabolism of unsaturated as well as saturated fatty acids. The results of this study will contriubte to a better understanding of PUFA metabolism and its regulation in normal and diseased tissues, e.g. in patients afflicted with Reye's syndrome.

本研究旨在回答一些尚未解决的问题 多不饱和脂肪酸（PUFA）和羟基脂肪酸的代谢 与白三烯有关。 为了充分了解其功能 过氧化物酶体 3-羟酰基-CoA 差向异构酶在 PUFA 降解中的作用，大鼠 肝酶将被纯化和表征。 特异性抑制剂 将设计 2,4-二烯酰辅酶 A 还原酶并缺乏缺乏的大肠杆菌突变体 将分离2,4-二烯酰辅酶A还原酶。 两者都是还原酶抑制剂 大肠杆菌突变体将用于确定 大鼠肝脏中 PUFA 降解的旧差向异构酶依赖性途径 过氧化物酶体和大肠杆菌。 2,4-二烯酰辅酶A还原酶抑制剂 也可用于确定还原酶催化的反应是否 PUFA 氧化限速。 存在或不存在 大鼠肝脏过氧化物酶体中的 cis-3-trans-2-enoyl-CoA 异构酶将 如果存在的话，酶将被分离和表征。 我们将继续研究2,4-二烯酰辅酶A的激素调节作用 PUFA 的还原酶和氧化作用。 这项研究需要 线粒体2,4-二烯酰辅酶A还原酶的纯化及其制备 抗还原酶抗体，将用于确定 mRNA 中的还原酶。 此外，利率之间的差异 顺式和反式不饱和脂肪酸的氧化将通过以下方法测定 大鼠心脏线粒体。 羟基脂肪酸的氧化包括 线粒体中的 5-羟基-6,8,11,13-二十碳四烯酸 (5-HETE) 和 将研究过氧化物酶体，以及对β-氧化的抑制 这些和其他一些潜在的抑制化合物。 的评估 β-氧化抑制剂对肝脏和心脏的有效性 需要酰基辅酶A合成酶的纯化和表征 存在于线粒体基质中。 最后，我们将测试3-癸炔酸 和其他可能的酰基辅酶A氧化酶抑制剂的有效性 作为过氧化物酶体脂肪酸氧化的抑制剂。 如果有效的话，这样的 化合物将用于评估过氧化物酶体的贡献 β-氧化对不饱和脂肪酸的肝脏代谢以及 饱和脂肪酸。 这项研究的结果将有助于 更好地了解 PUFA 代谢及其在正常和正常状态下的调节 患病组织，例如患有雷氏综合症的患者。