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. 2024 Jul 12;4(4):190–203. doi: 10.1021/acsbiomedchemau.4c00026

Figure 4.

Figure 4

Incorporation of unnatural amino acids into DCPs via amber codon suppression. (a) SDS-PAGE analysis of the Ni-NTA-purified His(6)-SUMO-MK1-3.6.10 fusion constructs with incorporated AzF at different positions. Green asterisks indicate the correct mass of desired full-length product. Red asterisks indicate observed truncation products. (b) Summary table of purified yield and measured affinities (via SPR) of azidophenylalanine-containing MK1-3.6.10 peptides compared to wild-type. n.b.: no binding; n.a.: not available. (c) Dimeric MK1-3.6.10 linked at the N-terminal AzF activates Wnt signaling in TOPbrite cells with a similar efficiency as a fully synthetic version dimerized at an N-terminal azidolysine group. (d) Crystal structure of ZNRF3-MK1-3.6.10 (PDB code: 8G4Y) complex showing a putative available surface for derivatization of F30. (e) Models of the extension of F30 aromatic moiety with either the azido group or the aminomethyl-triazole group.