Lactobionamide-based fluorinated detergent for functional and structural stabilization of membrane proteins
| Autor: | Romain Besson, Clarisse Faugier, Grégory Durand, Damien Cornut, Sébastien Igonet, Anass Jawhari |
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| Přispěvatelé: | Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Halogenation
[SDV]Life Sciences [q-bio] Detergents Druggability Lactobionamide Gene Expression Disaccharides General Biochemistry Genetics and Molecular Biology Receptors G-Protein-Coupled Surface-Active Agents 03 medical and health sciences Sf9 Cells Animals Molecular Biology 030304 developmental biology 0303 health sciences Chemistry Drug discovery Escherichia coli Proteins 030302 biochemistry & molecular biology Receptors Purinergic P1 Membrane Proteins Biological membrane Transporter Solubility Membrane protein Solubilization Chromatography Gel Biophysics Multidrug Resistance-Associated Proteins |
| Zdroj: | Methods Methods, Elsevier, 2020, 180, pp.19-26. ⟨10.1016/j.ymeth.2020.02.005⟩ |
| ISSN: | 1046-2023 1095-9130 |
| Popis: | Membrane proteins (MPs) are important drug discovery targets for a wide range of diseases. Conventional detergents such as n-Dodecyl β-D-maltoside have been used largely and efficiently to solubilize MPs with varying degrees of success concerning MPs functionality and stability. Fluorinated surfactants (FSs) have shown a great potential for the stabilization of various MPs. However, so far only a limited number of reports have demonstrated the ability of FSs to solubilize MPs from biological membranes. We report herein the use of a fluorinated lactobionamide-based detergent named FLAC6 for functional and structural stabilization of membrane proteins. We first demonstrated that FLAC6 efficiently solubilized three membrane proteins i.e. the native adenosine receptor A2AR, a G protein-coupled receptor, and two native transporters AcrB and BmrA. The resulting affinity purified MPs were highly pure, homogenous and aggregates free. Furthermore, the functionality of each MP was well maintained. Finally, striking overstabilization features were observed. Indeed, the Tm of native A2AR, AcrB and BmrA could be improved by 7, ~9 and ~ 23 °C, respectively when FLAC6 was used instead of the reference detergent. This work illustrates that FLAC6 is an efficient tool to maintain structural and functional integrities of different MPs belonging to different classes, providing a new avenue for functional stabilization of highly druggable and challenging membrane proteins involved in unmet medical needs. |
| Databáze: | OpenAIRE |
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