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Extracellular vesicles as a platform to study cell-surface membrane proteins
- 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.
- Subjects :
- Detergents
Protein-ligand interaction
GPI-Linked Proteins
Ligands
Extracellular vesicles
General Biochemistry, Genetics and Molecular Biology
Mass Spectrometry
law.invention
03 medical and health sciences
Extracellular Vesicles
Protein structure
law
Antibody generation
[CHIM.CRIS]Chemical Sciences/Cristallography
Humans
Cloning, Molecular
Molecular Biology
5'-Nucleotidase
030304 developmental biology
0303 health sciences
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
Chemistry
030302 biochemistry & molecular biology
Cryoelectron Microscopy
Membrane Proteins
Extracellular vesicle
Dynamic Light Scattering
Recombinant Proteins
Microscopy, Electron
Membrane
HEK293 Cells
Membrane protein
Solubilization
Biophysics
Recombinant DNA
Immunization
Cell surface membrane
Plasmids
Subjects
Details
- ISSN :
- 10959130 and 10462023
- Volume :
- 180
- Database :
- OpenAIRE
- Journal :
- Methods (San Diego, Calif.)
- Accession number :
- edsair.doi.dedup.....223a47bda24e6288844c0db7f81ce772