Fig. 5
From: Non-canonical amino acid labeling in proteomics and biotechnology
Overview of site-specific ncAA incorporation using orthogonal tRNA/aminoacyl synthetase pair. a A plasmid that expresses the desired orthogonal tRNA and tRNA synthetase is transfected into cells along with the plasmid containing the protein of interest that has been engineered to carry the suppressed codon sequence at a specific site. ncAA is added to the system and the protein of interest is labeled site-specfically with the ncAA. b An example of the codon sequence and corresponding peptides that result from either natural synthesis or synthesis in the presence of the orthogonal tRNA/tRNA synthetase and ncAA. c A peptide labeled site-specifically with a ncAA carrying an alkyne functional group is conjugated to a azide-containing fluorophore via CuAAC