Extended Data Fig. 8: Isolation and LC–MS analysis of P. putida tRNAIle2.
From: A tRNA modification with aminovaleramide facilitates AUA decoding in protein synthesis
a, Secondary structure of P. putida tRNAIle2 including modified nucleosides determined in this study by LC–MS. The RNase T1-digested fragment containing the anticodon is highlighted in red. b, Isolated P. putida tRNAIle2 was resolved by 10% PAGE with 7 M urea, stained with SYBR Gold and visualized by a FLA-7000 scanner (Fujifilm). c, LC–MS nucleoside analysis of the isolated tRNA. The total ion chromatogram (TIC; top), UV trace (second) and mass chromatograms of ava2C nucleoside (M + H)+ (m/z 342.18, third) and its base-related ion BH2+ (m/z 210.14, bottom) are shown. d, LC–MS analysis of P. putida tRNAIle2 digested by RNase T1. The BPC (top) and XICs of the anticodon-containing fragments bearing ava2C34 and t6A37 (middle) and ava2C34 and ct6A37 (bottom) are shown. RNA fragments with their m/z values and charge states (z) are shown in Supplementary Table 1e. A nonspecific peak with the same m/z was marked with an asterisk.
