Abstract
Membrane proteins (MPs) are stable in their native lipid environment. To enable structural and functional investigations, MPs need to be extracted from the membrane. This is a critical step that represents the main obstacle for MP biochemistry and structural biology. Here we describe detergent solubilization screening of MPs using dot-blot and Western-blot analyses. Good solubilization conditions are ranked for their best capacity to stabilize MPs using thermal shift assay. The protein functionality is evaluated by radioligand binding (for G-protein-coupled receptor) and ATPase activity (ABC Transporter) and finally the aggregation status as well as protein homogeneity are assessed by Native-polyacrylamide gel, chemical cross-linking, and size exclusion chromatography.
Key words
- Membrane proteins
- Solubilization
- Stabilization
- Detergent
- Native-PAGE
- Size exclusion chromatography
- Chemical-crosslinking
- Function
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Corvest, V., Jawhari, A. (2021). Solubilization and Stabilization of Native Membrane Proteins for Drug Discovery. In: Poterszman, A. (eds) Multiprotein Complexes. Methods in Molecular Biology, vol 2247. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1126-5_14
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DOI: https://doi.org/10.1007/978-1-0716-1126-5_14
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1125-8
Online ISBN: 978-1-0716-1126-5
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