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

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。

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.