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Allosteric regulation of G protein–coupled receptor activity by phospholipids

Nature Chemical Biology volume 12, pages 35–39 (2016)Cite this article

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Abstract

Lipids are emerging as key regulators of membrane protein structure and activity. These effects can be attributed either to the modification of bilayer properties (thickness, curvature and surface tension) or to the binding of specific lipids to the protein surface. For G protein–coupled receptors (GPCRs), the effects of phospholipids on receptor structure and activity remain poorly understood. Here we reconstituted purified β2-adrenergic receptor (β2R) in high-density lipoparticles to systematically characterize the effect of biologically relevant phospholipids on receptor activity. We observed that the lipid headgroup type affected ligand binding (agonist and antagonist) and receptor activation. Specifically, phosphatidylgycerol markedly favored agonist binding and facilitated receptor activation, whereas phosphatidylethanolamine favored antagonist binding and stabilized the inactive state of the receptor. We then showed that these effects could be recapitulated with detergent-solubilized lipids, demonstrating that the functional modulation occurred in the absence of a bilayer. Our data suggest that phospholipids act as direct allosteric modulators of GPCR activity.

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Figure 1: Lipids modulate ligand affinity of β2R.
Figure 2: β2R activation is regulated by phospholipids.
Figure 3: Modulation of β2R function by lipids does not require a bilayer.

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Acknowledgements

We thank R. Efremov for assistance with electron microscopy, E. Pardon and J. Steyaert (Vrije Universiteit Brussel) for providing purified Nb80, A. Manglik and D. Hilger for help with receptor production, L. Pardo for help with the statistical tests, R. Sunahara (University of California, San Diego) for providing the apoA-I plasmid and J.-M. Ruysschaert for helpful discussions. We acknowledge support from the Fonds de la Recherche Scientifique (FRS-FNRS) (grants F.4523.12, 34553.08 and T.0136.13), the US National Institute of Health (grants R01 NS028471 and R01GM083118) and a grant from the “Fond Extraordinaire de Recherche” (FER) 2007 of the Université Libre de Bruxelles. C.D. is a postdoctoral fellow of the FRS-FNRS. C.G. is a Chercheur Qualifié of the FRS-FNRS and a WELBIO investigator. M.M. was supported by a Hoover Foundation Brussels Fellowship from the Belgian American Educational Foundation (BAEF) and an American Heart Association (AHA) Award 15POST22700020.

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Authors and Affiliations

  1. Laboratory for the Structure and Function of Biological Membranes, Center for Structural Biology and Bioinformatics, Université Libre de Bruxelles, Brussels, Belgium

    Rosie Dawaliby, Cataldo Trubbia & Cédric Govaerts

  2. Laboratory of Pharmaceutical Chemistry Faculty of Pharmacy, Université Libre de Bruxelles, Brussels, Belgium

    Cédric Delporte & Pierre Van Antwerpen

  3. Analytical Platform of the Faculty of Pharmacy, Faculty of Pharmacy, Université Libre de Bruxelles, Brussels, Belgium

    Cédric Delporte & Pierre Van Antwerpen

  4. Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California, USA

    Matthieu Masureel & Brian K Kobilka

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  1. Rosie Dawaliby
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Contributions

R.D. expressed and purified the receptor and performed all HDL reconstitutions and functional characterizations, with the assistance of C.T. R.D. also performed all mass spectrometry analysis. M.M. also expressed and purified receptor samples. C.D. and P.V.A. performed mass spectrometry experiments. C.G. and B.K.K. provided overall project supervision and wrote the paper along with R.D.

Corresponding authors

Correspondence to Brian K Kobilka or Cédric Govaerts.

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The authors declare no competing financial interests.

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Dawaliby, R., Trubbia, C., Delporte, C. et al. Allosteric regulation of G protein–coupled receptor activity by phospholipids. Nat Chem Biol 12, 35–39 (2016). https://doi.org/10.1038/nchembio.1960

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