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. 2021 May 17;49(19):10807–10817. doi: 10.1093/nar/gkab288

Figure 4.

Figure 4.

Expression of an N-methyl-peptide containing 9 MeAAs and 15 pAAs. (A) Affinities of MeAA-tRNA#X to EF-Tu using all-#2, all-#4, and uniform sets. The orange bands indicate the range of ΔG values of pAA-tRNAs determined in Figure 2A (−8.0 to −9.4 kcal/mol). (B) Schematic representation of the EF-Tu affinity-tuning strategy. An appropriate combination of MeAA and tRNA#2–4 generates uniform affinities of MeAA-tRNA#Xs to EF-Tu comparable to canonical pAA-tRNAs. (C) The codon table reprogrammed with 9 MeAAs and 15 pAAs used in this study. (D) Sequences of mRNA2 and P2 peptide. (E) MALDI-TOF-MS analysis of P2 expressed using each set of MeAA and tRNA#X pairs. Arrowhead shows the corresponding monoisotopic peak of P2 or P2*. MALDI-TOF mass spectrum of P2-pAA is also shown.