Figure 1.

Design of luminescence-based assay for detection of IVT efficiency of noncanonical amino acids. (A) Overall schematic for IVT of luminescent peptides. AcF = N-acetyl-l-phenylalanine. Open blue circles indicate the 5 different luminescent peptide sequences detailed in Figure 1B. (B) Amino acid sequences of luminescent peptide variants tested in the P1A-E scaffolds. RLU and K_D values were previously reported in Dixon et al. for complementation with LgBiT; dashed values were not reported.²⁵ (C) Luminescence of IVT reactions for each luminescent peptide variant, monitored over 2 h. (D) Luminescence of a dilution series of a P1E peptide standard, monitored over 2 h. (E) Luminescence of the P1E peptide standard at 1 h (from Figure 1D) has a linear correlation to peptide concentration.