Furuta S, Hayashi H, Hijikata M, Miyazawa S, Osumi T, Hashimoto T
Proc Natl Acad Sci U S A. 1986 Jan;83(2):313-7. doi: 10.1073/pnas.83.2.313.
We have isolated five cDNA clones for rat liver catalase (hydrogen peroxide:hydrogen peroxide oxidoreductase, EC 1.11.1.6). These clones overlapped with each other and covered the entire length of the mRNA, which had been estimated to be 2.4 kilobases long by blot hybridization analysis of electrophoretically fractionated RNA. Nucleotide sequencing was carried out on these five clones and the composite nucleotide sequence of catalase cDNA was determined. The 5' noncoding region contained 83 bases and was followed by 1581 bases of an open reading frame that encoded 527 amino acids. The 3' noncoding region was 831 bases long and contained long repeats of the unit AC. The amino acid sequence deduced from the nucleotide sequence of the cDNAs showed about 90% homology with the reported primary structure of bovine liver catalase. The molecular weight of rat liver catalase was calculated to be 59,758 from the predicted amino acid sequence. The amino acid residues in contact with the heme group are completely identical for bovine liver and rat liver catalases. The amino acid sequence at the COOH terminus was confirmed by the results of carboxypeptidase P treatment of the protein purified from rat liver in the presence of leupeptin. Rat liver catalase has no cleavable signal peptide for translocation of the enzyme into peroxisomes.
我们已经分离出了五个大鼠肝脏过氧化氢酶(过氧化氢:过氧化氢氧化还原酶,EC 1.11.1.6)的cDNA克隆。这些克隆相互重叠,覆盖了mRNA的全长,通过对电泳分离的RNA进行印迹杂交分析,估计该mRNA长度为2.4千碱基。对这五个克隆进行了核苷酸测序,并确定了过氧化氢酶cDNA的复合核苷酸序列。5'非编码区包含83个碱基,其后是一个1581个碱基的开放阅读框,编码527个氨基酸。3'非编码区长831个碱基,包含AC单位的长重复序列。从cDNA的核苷酸序列推导的氨基酸序列与报道的牛肝脏过氧化氢酶一级结构显示约90%的同源性。根据预测的氨基酸序列计算大鼠肝脏过氧化氢酶的分子量为59,758。牛肝脏和大鼠肝脏过氧化氢酶中与血红素基团接触的氨基酸残基完全相同。通过在亮抑酶肽存在下对从大鼠肝脏纯化的蛋白质进行羧肽酶P处理的结果,证实了COOH末端的氨基酸序列。大鼠肝脏过氧化氢酶没有用于将该酶转运到过氧化物酶体中的可切割信号肽。