Analysis of Brain Fatty Acid Metabolism and its Abnormality by Application of Monoclonal Antibody

应用单克隆抗体分析脑脂肪酸代谢及其异常

• 批准号: 60480143
• 负责人: TAKESHITA Masazumi
• 金额: $ 3.9万
• 依托单位: Nedical College of Oita
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480143/
• 关键词: Fatty acid elogation; Very-long-chain fatty acid elongation; Monoclonal antibody Arachidyl-CoA; Arachidonoyl-CoA; NADH-cytochrome b_5 reductase; Redio-liquid chromatography; ラジオ液体クロマトグラフィー; 副腎白質ジストロフィー症; チトクロム【b_5】; 体外免疫法; ミクロソーム; 極長鎖脂肪酸伸長; 脂肪

In the present project, weinvestigated reaction systems of long- and very- long-chaine fatty acid elongation in detail. We prepared monoclonal antibodies for the enzymes and cytochromes involved in the elongation system, and investigated their application of the analysis of fatty acid metabolise and its abnormality in brain. The major results obtained are:1) We decided the cDNA nucleotide sequence of NADH-cytochrome b_5 reductase which was involved in fatty acid elongation. We obtained monoclonal antibody possessing high titer for the reductase.2) Computer simulation analysis revealed that the existence of wto parallel pathways in the very-long-chain fatty acid elongation in brain microsomes.3) At least two separate condensing snzymes were involved in the elongation depending on the chain length of fatty acids.4) In the elongation of arachidoyl-CoA and arachidonoyl-CoA condensation reaction waw evidenced to be rate limiting, and stereospecific S-sydrogen from NADPH was transfered in the reduction step.5) We found an elongation condition to produce 26:0 and 26:1, which have been produced specifically in adrenoleukodystrophy patients. These results will give an approch for the cause and process of the demyelinationg disease.

在本项目中，我们对长链和超长链脂肪酸延伸反应体系进行了详细的研究。我们制备了与延长系统有关的酶和细胞色素的单抗，并研究了它们在分析脑内脂肪酸代谢及其异常方面的应用。主要研究结果如下：1)测定了与脂肪酸伸长有关的NADH-细胞色素b5还原酶的基因序列。2)计算机模拟分析表明，在脑微粒体超长链脂肪酸的伸长过程中存在WTO平行通路。3)根据脂肪酸的链长，至少有两个独立的缩合酶参与了超长链脂肪酸的伸长。4)花生四氢叶酸-辅酶A和花生四烯酸-辅酶A的伸长反应被证明是限速的，来自NADPH的立体特异的S-水杨酸在还原步骤中转移。5)我们找到了产生26：0和26：1的伸长条件，这两种缩合酶是肾上腺营养不良患者特有的。这些结果将为探讨脱髓鞘病的病因和发病过程提供新的思路。

项目成果

竹下正純: "「生化学実験講座・続」第8巻 血液" 東京化学同人, (1987)

竹下正纯：《‘生物化学实验教程续’第8卷血》东京化学同人，（1987）