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. 2021 Oct 22;9:768854. doi: 10.3389/fchem.2021.768854

FIGURE 5.

FIGURE 5

Vc1c-mediated protein labelling. (A) Schematic of Vc1c-mediated ligation of DARPin (generated from PDB code 2QYJ) with fluorescent peptide RE(EDANS)AN-GI. A fluorescent peptide RE(EDANS)AN-GI was synthesized and DARPin9_26 was co-expressed with an N-terminal Gly-Leu dipeptide. The ligase can recognize and cleave the C-terminal Gly-Ile of the fluorescent peptide, and ligate it to the GL-DARPin9_26 N terminus. (B) Left panel: SDS-PAGE gel after Coomassie blue staining showing bands for DARPin9_26 around 15 kDa; Right: SDS-PAGE gel under UV illumination. The reaction was performed at 37°C, pH 6.5 with 20 nM Vc1c and 4 μM DARPin9_26.