Issue 4, 2021

Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes

Abstract

Templated chemistry offers the prospect of addressing specificity challenges occurring in bioconjugation reactions. Here, we show two peptide-templated amide-bond forming reactions that enable the concurrent labelling of two different membrane proteins with two different peptide nucleic acid (PNA) barcodes. The reaction system is based on the mutually selective coiled coil interaction between two thioester-linked PNA–peptide conjugates and two cysteine peptides serving as genetically encoded peptide tags. Orthogonal coiled coil templated covalent labelling is highly specific, quantitative and proceeds within a minute. To demonstrate the usefulness, we evaluated receptor internalisation of two membranous receptors EGFR (epidermal growth factor) and ErbB2 (epidermal growth factor receptor 2) by first staining PNA-tagged proteins with fluorophore–DNA conjugates and then erasing signals from non-internalized receptors via toehold-mediated strand displacement.

Graphical abstract: Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes

Supplementary files

Article information

Article type
Paper
Submitted
07 iyn 2021
Accepted
14 iyl 2021
First published
16 iyl 2021
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2021,2, 1291-1295

Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes

G. C. Gavins, K. Gröger, M. Reimann, M. D. Bartoschek, S. Bultmann and O. Seitz, RSC Chem. Biol., 2021, 2, 1291 DOI: 10.1039/D1CB00126D

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