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. 2022 Apr 29;10:873906. doi: 10.3389/fbioe.2022.873906

FIGURE 1.

FIGURE 1

Cartoon of amber suppression, reporter proteins, and ncAA sites. (A) Incorporation of a noncanonical amino acid (ncAA) requires three components: The suppressor tRNA is amino-acylated with the ncAA by the aminoacyl tRNA synthetase (aaRS). The ncAA-loaded suppressor tRNA recognizes the UAG (amber) stop codon of the mRNA and the ribosome incorporates the ncAA into the nascent peptide chain leading to a suppression product containing a ncAA. Translation termination competes with ncAA incorporation due to codon stop recognition by release factor eRF1, causing the generation of a termination product not containing a ncAA. (B) Schematic 3D-structure of the reporter proteins RFP-Y-GFP and RFP-PNT-GFP (left, the yellow circles depict the ncAA incorporation sites) and schematic of respective DNA sequences (right, the yellow bars with arrows depict the ncAA incorporation sites). 3D structure cartoons are based on PDB structures 2H5Q, 5SX4, and 2B3P.