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

Authors :
Magali Grange
Vincent Delauzun
Laurent Gauthier
Alain Roussel
Philippe Leone
Anaïs Gaubert
Béatrice Amigues
Architecture et fonction des macromolécules biologiques (AFMB)
Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Innate Pharma Research Labs [Marseille]
Innate Pharma
ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010)
ANR-14-EBOL-0001,STOP-EBOLA,Neutralization du virus Ebola par des nanobodies de camélidés(2014)
Source :
Methods, Methods, Elsevier, 2020, 180, pp.35-44. ⟨10.1016/j.ymeth.2020.03.004⟩, Methods, 2020, 180, pp.35-44. ⟨10.1016/j.ymeth.2020.03.004⟩
Publication Year :
2019

Abstract

International audience; 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.

Details

ISSN :
10959130 and 10462023
Volume :
180
Database :
OpenAIRE
Journal :
Methods (San Diego, Calif.)
Accession number :
edsair.doi.dedup.....223a47bda24e6288844c0db7f81ce772