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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Nucleotide sequence of medium-chain acyl-CoA dehydrogenase mRNA and its expression in enzyme-deficient human tissue.
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Nucleotide sequence of medium-chain acyl-CoA dehydrogenase mRNA and its expression in enzyme-deficient human tissue.

机译：中链酰基辅酶A脱氢酶mRNA的核苷酸序列及其在缺酶人体组织中的表达。

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摘要

Medium-chain acyl-CoA dehydrogenase (MCAD; acyl-CoA: (acceptor) 2,3-oxidoreductase, EC 1.3.99.3) is one of three similar enzymes that catalyze the initial step of fatty acid beta-oxidation. Definition of the primary structure of MCAD and the tissue distribution of its mRNA is of biochemical and clinical importance because of the recent recognition of inherited MCAD deficiency in humans. The MCAD mRNA nucleotide sequence was determined from two overlapping cDNA clones isolated from human liver and placental cDNA libraries, respectively. The MCAD mRNA includes a 1263-base-pair coding region and a 738-base-pair 3'-nontranslated region. A partial amino acid sequence (137 residues) determined on peptides derived from MCAD purified from porcine liver confirmed the identity of the cDNA clone. Comparison of the amino acid sequence predicted from the human MCAD cDNA with the partial protein sequence of the porcine MCAD revealed a high degree (88%) of interspecies sequence identity. RNA blot analysis shows that MCAD mRNA is expressed in a variety of rat (2.2 kilobases) and human (2.4 kilobases) tissues. Blot hybridization of RNA prepared from cultured skin fibroblasts from a patient with MCAD deficiency disclosed that mRNA was present and of similar size to MCAD mRNA derived from control fibroblasts. The isolation and characterization of MCAD cDNA is an important step in the definition of the defect underlying MCAD deficiency and in understanding its metabolic consequences.
机译：中链酰基辅酶A脱氢酶（MCAD;酰基辅酶A：（受体）2,3-氧化还原酶，EC 1.3.99.3）是催化脂肪酸β-氧化起始步骤的三种相似酶之一。 MCAD的主要结构及其mRNA的组织分布的定义具有生化和临床意义，因为最近认识到人类遗传性MCAD缺乏症。从分别从人肝脏和胎盘cDNA文库中分离出的两个重叠cDNA克隆中确定MCAD mRNA核苷酸序列。 MCAD mRNA包括一个1263个碱基对的编码区和一个738个碱基对的3'非翻译区。在从猪肝脏纯化的MCAD衍生的肽上测定的部分氨基酸序列（137个残基）证实了cDNA克隆的身份。从人MCAD cDNA预测的氨基酸序列与猪MCAD的部分蛋白序列的比较表明，物种间的序列高度一致（88％）。 RNA印迹分析表明，MCAD mRNA在多种大鼠（2.2千碱基）和人（2.4千碱基）组织中表达。从患有MCAD缺乏症患者的培养的皮肤成纤维细胞制备的RNA的印迹杂交显示存在mRNA，并且大小与衍生自对照成纤维细胞的MCAD mRNA相似。 MCAD cDNA的分离和表征是定义MCAD缺陷基础缺陷和了解其代谢后果的重要步骤。

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