Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Jan;12(1):35-9.
doi: 10.1038/nchembio.1960. Epub 2015 Nov 16.

Allosteric regulation of G protein-coupled receptor activity by phospholipids

Rosie Dawaliby  1 , Cataldo Trubbia  1 , Cédric Delporte  2   3 , Matthieu Masureel  4 , Pierre Van Antwerpen  2   3 , Brian K Kobilka  4 , Cédric Govaerts  1
Affiliations

Allosteric regulation of G protein-coupled receptor activity by phospholipids

Rosie Dawaliby et al. Nat Chem Biol. 2016 Jan.

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.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Lipids modulate ligand affinity of β2R
a. Chemical structure of the lipids used for in this study. For clarity the acyl chains are not shown and replaced by R1 and R2 labels. b-c. Ligand binding curves for the agonist Isoproterenol and the antagonist Alprenolol competing against [3H]-dihydroalprenolol ([3H]-DHA) for β2R reconstituted in rHDL particles of different lipid compositions.
Figure 2
Figure 2. β2R activation is regulated by phospholipids
a. Molecular surface representation of the active (PDB id: 3SN6) and inactive (PDB id: 4GBR) structures of the β2R with TM6 highlighted (colored ribbon) and the position of Cys265 (in yellow), showing the increased exposure of the latter in the active state. b. Wavelength of the emission maximum fluorescence (λmax) in the absence of ligand (basal) and after addition of 100μM of inverse agonist ICI-118,551 for bimane-labelled receptors reconstituted in rHDLs of different lipid compositions compared with DDM solubilized receptor. Asterisk denotes statistically significant difference (p<0.05). c. Basal fluorescence emission spectra for bimane-labelled receptors in rHDL of DOPC, DOPG and DOPE in comparison with DDM solubilized receptor. The effect of the inverse agonist (ICI-118,551) on β2R in DOPG rHDL is represented by dashed red curve. Blue shift in λmax is highlighted by an arrow. d. Time dependent fluorescence change on β2R rHDL in DOPC upon addition of saturating concentration of 200μM agonist (Isoproterenol). The λmax positions are highlighted on each curve. e. Fluorescence recovery of agonist-stimulated receptor shown in d upon the addition of saturating 100μM antagonist (Alprenolol). f. Time dependent changes in bimane λmax upon addition of Isoproterenol (200 μM) followed by Alprenolol (100μM) for labelled receptor reconstituted in the different rHDLs. Data are the mean ± SEM of at least 3 independent experiments.g. Time dependent changes in bimane λmax upon addition of 4μM Nb80 followed by agonist activation (200μM Isoproterenol). Data are the mean ± SEM of at least 3 independent experiments.
Figure 3
Figure 3. Modulation of β2R function by lipids does not require a bilayer
a. Time dependent changes in bimane λmax of solubilized bimane-labelled β2R (1μM receptor, 500μM DDM) upon addition of saturating concentration of 200μM Isoproterenol in the presence of increasing concentration of solubilized DOPG (with 600 μM Sodium Cholate). No lipids condition was performed in presence of 600 μM sodium cholate. Data are the mean ± SEM of at least 3 independent experiments. b. Time dependent changes in bimane λmax upon addition of saturating concentration of 200μM Isoproterenol in the presence of 200 μM of either DOPC, DOPG, DOPE and DOPS (detergent concentrations are identical to those in panel a). Data are the mean ± SEM of at least 3 independent experiments. c. Time dependent changes in bimane λmax upon addition of 4μM Nb80 followed by agonist activation (200μM Isoproterenol). Data are the mean ± SEM of at least 3 independent experiments.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Lee AG. Biological membranes: the importance of molecular detail. Trends Biochem. Sci. 2011;36:493–500. - PubMed
    1. Gourdon P, et al. HiLiDe: Systematic Approach to Membrane Protein Crystallization in Lipid and Detergent. Crystal Growth & Design. 2011;11:2098–2106.
    1. Lee AG. How lipids affect the activities of integral membrane proteins. Biochim. Biophys. Acta. 2004;1666:62–87. - PubMed
    1. Laganowsky A, et al. Membrane proteins bind lipids selectively to modulate their structure and function. Nature. 2014;510:172–175. - PMC - PubMed
    1. Hunte C, Richers S. Lipids and membrane protein structures. Curr. Opin. Struct. Biol. 2008;18:406–411. - PubMed

Publication types

MeSH terms

Substances