Furan Cross-Linking Technology for Investigating GPCR–Ligand Interactions

  • Marleen Van Troys
  • Willem Vannecke
  • Christophe Ampe
  • Annemieke MadderEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1947)


Interactions between G protein-coupled receptors and their ligands hold extensive potential for drug discovery. Studying these interactions poses technical problems due to their transient nature and the inherent difficulties when working with G protein-coupled receptors (GPCR) that are only functional in a membrane setting. Here, we describe the use of a furan-based chemical cross-linking methodology to achieve selective covalent coupling between a furan-modified peptide ligand and its native GPCR present on the surface of living cells under normal cell culture conditions. This methodology relies on the oxidation of the furan moiety, which can be achieved by either addition of an external oxidation signal or by the reactive oxygen species produced by the cell. The cross-linked ligand–GPCR complex is subsequently detected by Western blotting based on the biotin label that is incorporated in the peptide ligand.

Key words

Furan Receptor GPCR Chemical cross-linking Peptide Solid phase peptide synthesis Western blot Singlet oxygen Reactive oxygen species 


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Marleen Van Troys
    • 1
    • 2
  • Willem Vannecke
    • 3
  • Christophe Ampe
    • 1
    • 2
  • Annemieke Madder
    • 3
    Email author
  1. 1.Department of Biochemistry, Faculty of Medicine and Health SciencesGhent UniversityGhentBelgium
  2. 2.Department of Biomolecular Medicine, Faculty of Medicine and Health SciencesGhent UniversityGhentBelgium
  3. 3.Organic and Biomimetic Chemistry Research Group, Department of Organic and Macromolecular ChemistryGhent UniversityGhentBelgium

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