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The processing of human mitochondrial leucyl‐tRNA synthetase in the insect cells
Author(s) -
Yao Yong-Neng,
Wang Lie,
Wu Xiang-Fu,
Wang En-Duo
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)03833-4
Subject(s) - aquifex aeolicus , mitochondrion , transfer rna , biochemistry , escherichia coli , complementary dna , enzyme , biology , signal peptide , microbiology and biotechnology , peptide sequence , rna , gene
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.