OaAEP1-mediated PNA-protein conjugation enables erasable imaging of membrane proteins

We report the use of a protein ligase to covalently ligate a protein to a peptide nucleic acid (PNA). The rapid ligation demands only an N-terminal GL dipeptide in the target protein and a C-terminal NGL tripeptide in the PNA. We demonstrate the versatility of this approach by attaching a PNA strand to three different proteins. Lastly, we show that erasable imaging of EGFR on HEK293 cell membranes is achieved with DNA origami nanostructures and toehold-mediated strand displacement.

Automated summary

We describe the use of a protein ligase for covalently attaching a protein to a peptide nucleic acid (PNA). This approach only requires a GL dipeptide at the N-terminus of the target protein and an NGL tripeptide at the C-terminus of the PNA. Our study demonstrates the versatility of this method by successfully attaching a PNA strand to three different proteins. Additionally, we show the feasibility of erasable imaging of EGFR on HEK293 cell membranes using DNA origami nanostructures and toehold-mediated strand displacement.