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Amphipols in G Protein-Coupled Receptor Pharmacology: What Are They Good For?

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Abstract

G protein-coupled receptors are at a central node of all cell communications. Investigating their molecular functioning is therefore crucial for both academic purposes and drug design. However, getting the receptors as isolated, stable and purified proteins for such studies still stumbles over their instability out of the membrane environment. Different membrane-mimicking environments have been developed so far to increase the stability of purified receptors. Among them are amphipols. These polymers not only preserve the native fold of receptors purified from membrane fractions but they also allow specific applications such as folding receptors purified from inclusion bodies back to their native state. Of importance, amphipol-trapped G protein-coupled receptors essentially maintain their pharmacological properties so that they are perfectly adapted to further investigate the molecular mechanisms underlying signaling processes. We review here how amphipols have been used to refold and stabilize detergent-solubilized purified receptors and what are the main subsequent molecular pharmacology analyses that were performed using this strategy.

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Abbreviations

5HT4(a):

5-Hydroxytryptamine receptor 4

12-HHT:

12S-Hydroxyheptadeca-5Z, 8E,10E-trienoic acid

A8-35:

Polyacrylate-based amphipol A8-35

AVP:

Arginine-vasopressin

BLT1:

Leukotriene B4 receptor 1

BLT2:

Leukotriene B4 receptor 2

CB1:

Cannabinoid receptor 1

CRINEPT:

Cross-correlated relaxation-enhanced polarization transfer

CXCR1:

C-X-C chemokine receptor 1

DDM:

Dodecyl-β-D-maltopyranoside

FRET:

Fluorescence resonance energy transfer

FomA:

Outer membrane protein A from Fusobacterium nucleatum

GHS-R1a:

Ghrelin receptor type 1

GPCR:

G protein-coupled receptor

IB:

Inclusion bodies

LRET:

Luminescence resonance energy transfer

LTB4 :

Leukotriene B4

MNG:

Maltose-neopentylglucose

OmpA:

Outer membrane protein A from Escherichia coli

NaPol:

Non-ionic amphipol

NMR:

Nuclear magnetic resonance

Sf9 :

Spodoptera frugiperda

V2R:

Vasopressin receptor 2

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Acknowledgments

We are particularly indebted to J.-L. Popot (IBPC, Paris) for his invaluable contribution to all this work. We also wish to thank L. Catoire and M. Zoonens (IBPC, Paris) for helpful discussions. This work was supported by CNRS and National Agency of Research Grants ANR 06-BLAN-0087, ANR-10-BLAN-1208 and PCV08_323163.

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Authors and Affiliations

  1. Faculté de Pharmacie, Institut des Biomolécules Max Mousseron (IBMM), CNRS UMR 5247, Université Montpellier 1 et 2, BP 14491, 15 Avenue Charles Flahault, 34093, Montpellier Cedex 5, France

    Sophie Mary, Marjorie Damian, Jacky Marie & Jean-Louis Banères

  2. Institut de Génomique Fonctionnelle (IGF), CNRS UMR 5203, INSERM U661, Université Montpellier 1 et 2, 141 rue de la Cardonille, 34094, Montpellier Cedex 05, France

    Rita Rahmeh, Bernard Mouillac & Sébastien Granier

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  1. Sophie Mary
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  2. Marjorie Damian
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  3. Rita Rahmeh
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  4. Bernard Mouillac
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  6. Sébastien Granier
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  7. Jean-Louis Banères
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Correspondence to Jean-Louis Banères.

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Mary, S., Damian, M., Rahmeh, R. et al. Amphipols in G Protein-Coupled Receptor Pharmacology: What Are They Good For?. J Membrane Biol 247, 853–860 (2014). https://doi.org/10.1007/s00232-014-9665-9

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  • DOI: https://doi.org/10.1007/s00232-014-9665-9

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