Osumi T, Ishii N, Hijikata M, Kamijo K, Ozasa H, Furuta S, Miyazawa S, Kondo K, Inoue K, Kagamiyama H
J Biol Chem. 1985 Jul 25;260(15):8905-10.
For the studies on the mechanism of induction of peroxisomal beta-oxidation enzymes and biogenesis of the organelle, we have isolated cDNA clones for rat peroxisomal enoyl-CoA: hydratase-3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme. On blotting experiments with liver RNA, the cDNAs hybridized to a 3.0-kilobase RNA which was increased 5-7-fold by the administration of di-(2-ethylhexyl)phthalate to rats. Nucleotide sequencing was carried out for four cloned cDNAs and one obtained by a primer extension method. By overlapping these sequences with each other, we identified 20 nucleotides of 5'-noncoding, 2,166 nucleotides of coding, and 910 nucleotides of 3'-noncoding regions. The deduced amino acid sequence of the enzyme is composed of 722 residues, and the composition agrees with that of the protein data. The sequence was confirmed by the amino acid compositions and sequence analyses of some of the tryptic peptides. The molecular weight of the mature enzyme is calculated to be 78,511 from the predicted amino acid sequence. The enzyme has no terminal peptide extension as a signal for translocation into peroxisomes.
为了研究过氧化物酶体β-氧化酶的诱导机制和该细胞器的生物发生，我们分离了大鼠过氧化物酶体烯酰辅酶A：水化酶-3-羟酰基辅酶A脱氢酶双功能酶的cDNA克隆。在用肝脏RNA进行的印迹实验中，这些cDNA与一个3.0千碱基的RNA杂交，该RNA在给大鼠施用邻苯二甲酸二（2-乙基己基）酯后增加了5至7倍。对四个克隆的cDNA和一个通过引物延伸法获得的cDNA进行了核苷酸测序。通过将这些序列相互重叠，我们确定了5'-非编码区的20个核苷酸、编码区的2166个核苷酸和3'-非编码区的910个核苷酸。该酶推导的氨基酸序列由722个残基组成，其组成与蛋白质数据一致。通过对一些胰蛋白酶肽段的氨基酸组成和序列分析证实了该序列。根据预测的氨基酸序列计算出成熟酶的分子量为78,511。该酶没有作为转运到过氧化物酶体信号的末端肽延伸。
