Orthogonal coiled coils enable rapid covalent labelling of two distinct membrane proteins with peptide nucleic acid barcodes.
| Autor: | Gavins GC; Institut für Chemie, Humboldt-Universität zu Berlin Brook-Taylor-Strasse 2 Berlin 12489 Germany oliver.seitz@chemie.hu-berlin.de., Gröger K; Institut für Chemie, Humboldt-Universität zu Berlin Brook-Taylor-Strasse 2 Berlin 12489 Germany oliver.seitz@chemie.hu-berlin.de., Reimann M; Institut für Chemie, Humboldt-Universität zu Berlin Brook-Taylor-Strasse 2 Berlin 12489 Germany oliver.seitz@chemie.hu-berlin.de., Bartoschek MD; Center for Molecular Biosystems (BioSysM), Faculty of Biology, Ludwig-Maximilians-Universität München, Butenandtstr. 1 Munich 81377 Germany bultmann@bio.lmu.de., Bultmann S; Center for Molecular Biosystems (BioSysM), Faculty of Biology, Ludwig-Maximilians-Universität München, Butenandtstr. 1 Munich 81377 Germany bultmann@bio.lmu.de., Seitz O; Institut für Chemie, Humboldt-Universität zu Berlin Brook-Taylor-Strasse 2 Berlin 12489 Germany oliver.seitz@chemie.hu-berlin.de. |
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| Jazyk: | angličtina |
| Zdroj: | RSC chemical biology [RSC Chem Biol] 2021 Jul 16; Vol. 2 (4), pp. 1291-1295. Date of Electronic Publication: 2021 Jul 16 (Print Publication: 2021). |
| DOI: | 10.1039/d1cb00126d |
| Abstrakt: | 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. Competing Interests: There are no conflicts to declare. (This journal is © The Royal Society of Chemistry.) |
| Databáze: | MEDLINE |
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