Extracellular vesicles as a platform to study cell-surface membrane proteins

Vincent Delauzun (1) , Beatrice Amigues (1) , Anais Gaubert (1) , Philippe Leone (1) , Magali Grange (1) , Laurent Gauthier (2) , Alain Roussel (1)

1 AFMB - Architecture et fonction des macromolécules biologiques
2 Innate Pharma Research Labs [Marseille]

Keywords Extracellular vesicle production Membrane protein expression Membrane protein-ligand interaction Cell-surface receptor Electron microscopy Immunization Highlights Membrane protein interaction studies using extracellular vesicles Extracellular vesicles as a tool for structure determination of membrane proteins Extracellular vesicles use for immunization Abstract Producing intact recombinant membrane proteins for structural studies is an inherently challenging task due to their requirement for a cell-lipid environment. Most procedures developed involve isolating the protein by detergent solubilization and further reconstitutions into artificial membranes. These procedures are highly time consuming and suffer from further drawbacks, including low yields and high cost. We describe here an alternative method for rapidly obtaining recombinant cell-surface membrane proteins displayed on extracellular vesicles (EVs) derived from cells in culture. Interaction between these membrane proteins and ligands can be analyzed directly on EVs. Moreover, EVs can also be used for protein structure determination or immunization purposes.

Mots clés: Extracellular vesicle; Membrane protein; Protein-ligand interaction; Immunization; Antibody generation

Domaines: Cristallographie; Biologie structurale [q-bio.BM]

Vincent Delauzun, Beatrice Amigues, Anais Gaubert, Philippe Leone, Magali Grange, et al.. Extracellular vesicles as a platform to study cell-surface membrane proteins. Methods, 2020, 180, pp.35-44. ⟨10.1016/j.ymeth.2020.03.004⟩. ⟨hal-02989372⟩