Solubilization and Stabilization of Native Membrane Proteins for Drug Discovery.

Vincent Corvest, Anass Jawhari

Methods in molecular biology (Clifton, N.J.) 2021

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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.

Citation

Vincent Corvest, Anass Jawhari. Solubilization and Stabilization of Native Membrane Proteins for Drug Discovery. Methods in molecular biology (Clifton, N.J.). 2021;2247:257-267