Optimal Codon Choice Can Improve the Efficiency and Fidelity of N-Methyl Amino Acid Incorporation into Peptides by In-Vitro Translation

摘要

The generation of peptides containing N-methyl amino acids (N-Me AAs) by in-vitro translation is of interest as a potential route to the directed evolution of pharmaceutically useful modified peptides. However, N-Me AAs are suboptimal substrates for the translational apparatus, and as a result, premature termination of translation can occur or incorrect amino acids can be misincorporated in place of the desired N-Me AA, resulting in reduced incorporation efficiency and fidelity of the N-Me AA. We sought to determine the main cause of such misincorporation and truncation events, and then to optimize the incorporation of N-Me AAs.