The processing of human mitochondrial leucyl‐tRNA synthetase in the insect cells

FEBS Letters

Wang, Lie¹  Yao, Yong-Neng²  Wang, En-Duo²  Wu, Xiang-Fu¹ 

[1] Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, PR China;State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, PR China

关键词: Human mitochondrion;    Aminoacyl-tRNA synthetase;    tRNALeu;    Baculovirus;    Aquifex aeolicus;    Escherichia coli;    aaRS;    aminoacyl-tRNA synthetase;    LeuRS;    leucyl-tRNA synthetase;    hmLeuRS;    human mitochondrial leucyl-tRNA synthetase;    hmtRNALeu;    human mitochondrial tRNALeu;    Tn5;    Trichoplusia ni BTI-Tn-5B1-4;    SDS–PAGE;    sodium dodecyl sulfate–polyacrylamide gel electrophoresis;   

DOI  :  10.1016/S0014-5793(02)03833-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A His-tagged full-length cDNA of human mitochondrial leucyl-tRNA synthetase was expressed in a baculovirus system. The N-terminal sequence of the enzyme isolated from the mitochondria of insect cells was found to be IYSATGKWTKEYTL, indicating that the mitochondrial targeting signal peptide was cleaved between Ser39 and Ile40 after the enzyme precursor was translocated into mitochondria. The enzyme purified from mitochondria catalyzed the leucylation of Escherichia coli tRNA₁ ^Leu(CAG) and Aquifex aeolicus tRNA^Leu(GAG) with higher catalytic activity in the leucylation of E. coli tRNA^Leu than that previously expressed in E. coli without the N-terminal 21 residues.

【 授权许可】

Unknown