The processing of human mitochondrial leucyl-tRNA synthetase in the insect cells.
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 tRNA(1)(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.
Yao
Yong Neng
YN
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.
Wang
Lie
L
Wu
Xiang Fu
XF
Wang
En Duo
ED
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D006801:Humans; D007915:Lepidoptera / Q000166:cytology / Q000235:genetics; D007935:Leucine-tRNA Ligase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011498:Protein Precursors / Q000378:metabolism; D021381:Protein Transport; D012356:RNA, Transfer, Leu / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D029968:Escherichia coli Proteins; D010446:Peptide Fragments; D011498:Protein Precursors; D012356:RNA, Transfer, Leu; D011994:Recombinant Proteins; D006639:Histidine; D007935:Leucine-tRNA Ligase