Fig. 4: Further characterization of PylRS variants. | Nature Communications

Fig. 4: Further characterization of PylRS variants.

From: Machine learning-guided evolution of pyrrolysyl-tRNA synthetase for improved incorporation efficiency of diverse noncanonical amino acids

Fig. 4

a Introduction of 3MetH at His93 position of myoglobin as a ligand of heme. b Product yield of reaction catalyzed by myoglobin-3MetH after 40-min reaction, and expression of myoglobin-3MetH detected on SDS-PAGE. The incorporation of 3MetH was enabled by IFGFF and Com2-IFGFF. +, with 3MetH added; −, no 3MetH added. Myoglobin-WT indicates no ncAA incorporation in myoglobin. c Suppression of multiple amber codons by PylRS variants, with multiple consecutive amber codons inserted at the second position of sfGFP. d Fluorescence intensity of sfGFP with multiple 3BrF inserted at the second position, enabled by IFRS, Com1-IFRS, and Com2-IFRS. e The positions where multiple 3BrF are inserted in sfGFP. f Fluorescence intensity of sfGFP with multiple 3BrF inserted at different positions, enabled by IFRS, Com1-IFRS, and Com2-IFRS. Error bars represent ±standard deviation of the mean over four independent replicates. Source data are provided as a Source Data file.

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