Figure 3.

Substrate specificities of Mx1201 PylRS/PylT and Mx1201 PylRS^Y126A/PylT. Fluorescence plate reader assay from HEK293T cell lysates transiently transfected in a 4:1 ratio with a GFP^150TAG reporter and (a) Mma PylRS/PylT, (b) Mx1201 PylRS/PylT, (c) Mx1201 PylRS^Y126A/PylT, or (d) Mma PylRS AF/PylT. GFP fluorescence is shown as a percentage of fluorescence measured with a GFP construct without a TAG stop codon in the same experiment. Cells were grown for 24 h in the absence (-ncAA) or presence of one of the following ncAAs: 0.2 mM CpK, 0.5 mM BCNK, 0.1 mM TCO*K, 0.5 mM AbK, 0.5 mM PcK, and 10 mM AcK. For full names and structures of amino acids, refer to Supporting Information Figure 3.