Skip to main content
. 2022 Dec 1;299(1):102759. doi: 10.1016/j.jbc.2022.102759

Figure 6.

Figure 6

TrmK recognizes the U8, A14, G18, and U55 in tRNALeu.a and b, representative tertiary interactions of (A) U8-A14 and (B) G18-U55 from the complex of Thermus thermophilus Leucyl-tRNA synthetase with tRNALeu transcript (PDB: 2BYT) are illustrated. The nucleotides in the tRNALeu variants where methyl group acceptance activities (mutation rates at A22) less than 0.05 are highlighted in red. C, the methyl group acceptance activities in tRNALeu_GAG_2 WT, U8A, U8A-A14U, G18U, and G18U-U55G were measured with a gel assay. The TrmK reaction was performed in the reaction mixture containing 50 mM Tris–HCl (pH 7.6), 200 mM KCl, 5 mM MgCl2, 0.0125 OD/μl tRNA, 50 μM 14C-S-adenosyl-L-methionine (AdoMet), and 100 mM TrmK at 60 °C for 10 min. The gel was stained with Methylene blue (MB), and 14C-methyl group incorporation was monitored by autoradiography.