Affinity labelling of E. coli leucyl-tRNA synthetase with 3'-oxidized tRNA(Leu).
The E. coli leucyl-tRNA synthetase (E.C. 6.1.1.4) was specifically labelled with 3'-oxidized tRNA(Leu) (tRNA(oxLeu)). The procedure involves a Schiff's base formation and its subsequent reduction by sodium cyanoborohydride. Stoichiometric inactivation of aminoacylation was achieved with the incorporation of 1 mol of tRNA(oxLeu) per mol LeuRS. On the other hand, the amino acid activation activity of LeuRS-tRNA(ox) complex was partially inhibited. After extensive digestion of the complex by pancreatic ribonuclease, the amino acid activation activity was fully recovered, while the aminoacylation activity was not restored at all.
D000345:Affinity Labels; D000604:Amino Acyl-tRNA Synthetases; D004926:Escherichia coli / Q000201:enzymology*; D007935:Leucine-tRNA Ligase; D012329:RNA, Bacterial; D012345:RNA, Transfer, Amino Acid-Specific; D012356:RNA, Transfer, Leu
D000345:Affinity Labels; D012329:RNA, Bacterial; D012345:RNA, Transfer, Amino Acid-Specific; D012356:RNA, Transfer, Leu; D000604:Amino Acyl-tRNA Synthetases; D007935:Leucine-tRNA Ligase