Membrane Cholesterol Access into a G-protein-coupled Receptor

Overview

Journal Nat Commun

Specialty Biology

Date 2017 Feb 22

PMID 28220900

Citations 73

Authors

Ramon Guixa-Gonzalez

Jose L Albasanz

Ismael Rodriguez-Espigares

Manuel Pastor

Ferran Sanz

Maria Marti-Solano

Moutusi Manna

Hector Martinez-Seara

Peter W Hildebrand

Mairena Martin

Jana Selent

Affiliations

Soon will be listed here.

Abstract

Cholesterol is a key component of cell membranes with a proven modulatory role on the function and ligand-binding properties of G-protein-coupled receptors (GPCRs). Crystal structures of prototypical GPCRs such as the adenosine A receptor (AR) have confirmed that cholesterol finds stable binding sites at the receptor surface suggesting an allosteric role of this lipid. Here we combine experimental and computational approaches to show that cholesterol can spontaneously enter the AR-binding pocket from the membrane milieu using the same portal gate previously suggested for opsin ligands. We confirm the presence of cholesterol inside the receptor by chemical modification of the AR interior in a biotinylation assay. Overall, we show that cholesterol's impact on AR-binding affinity goes beyond pure allosteric modulation and unveils a new interaction mode between cholesterol and the AR that could potentially apply to other GPCRs.

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References

Nachtergaele S, Mydock L, Krishnan K, Rammohan J, Schlesinger P, Covey D . Oxysterols are allosteric activators of the oncoprotein Smoothened. Nat Chem Biol. 2012; 8(2):211-20. PMC: 3262054. DOI: 10.1038/nchembio.765. View

Wu H, Wang C, Gregory K, Han G, Cho H, Xia Y . Structure of a class C GPCR metabotropic glutamate receptor 1 bound to an allosteric modulator. Science. 2014; 344(6179):58-64. PMC: 3991565. DOI: 10.1126/science.1249489. View

Piechnick R, Ritter E, Hildebrand P, Ernst O, Scheerer P, Hofmann K . Effect of channel mutations on the uptake and release of the retinal ligand in opsin. Proc Natl Acad Sci U S A. 2012; 109(14):5247-52. PMC: 3325672. DOI: 10.1073/pnas.1117268109. View

Hildebrand P, Scheerer P, Park J, Choe H, Piechnick R, Ernst O . A ligand channel through the G protein coupled receptor opsin. PLoS One. 2009; 4(2):e4382. PMC: 2632885. DOI: 10.1371/journal.pone.0004382. View

Mitchell D, Straume M, Miller J, Litman B . Modulation of metarhodopsin formation by cholesterol-induced ordering of bilayer lipids. Biochemistry. 1990; 29(39):9143-9. DOI: 10.1021/bi00491a007. View

Park J, Morizumi T, Li Y, Hong J, Pai E, Hofmann K . Opsin, a structural model for olfactory receptors?. Angew Chem Int Ed Engl. 2013; 52(42):11021-4. DOI: 10.1002/anie.201302374. View

Zocher M, Zhang C, Rasmussen S, Kobilka B, Muller D . Cholesterol increases kinetic, energetic, and mechanical stability of the human β2-adrenergic receptor. Proc Natl Acad Sci U S A. 2012; 109(50):E3463-72. PMC: 3528585. DOI: 10.1073/pnas.1210373109. View

Niu S, Mitchell D, Litman B . Manipulation of cholesterol levels in rod disk membranes by methyl-beta-cyclodextrin: effects on receptor activation. J Biol Chem. 2002; 277(23):20139-45. DOI: 10.1074/jbc.M200594200. View

Colozo A, Park P, Sum C, Pisterzi L, Wells J . Cholesterol as a determinant of cooperativity in the M2 muscarinic cholinergic receptor. Biochem Pharmacol. 2007; 74(2):236-55. DOI: 10.1016/j.bcp.2007.04.009. View

10.

Jaakola V, Griffith M, Hanson M, Cherezov V, Chien E, Lane J . The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist. Science. 2008; 322(5905):1211-7. PMC: 2586971. DOI: 10.1126/science.1164772. View

11.

Wang T, Duan Y . Ligand entry and exit pathways in the beta2-adrenergic receptor. J Mol Biol. 2009; 392(4):1102-15. PMC: 2809666. DOI: 10.1016/j.jmb.2009.07.093. View

12.

Eswar N, Eramian D, Webb B, Shen M, Sali A . Protein structure modeling with MODELLER. Methods Mol Biol. 2008; 426:145-59. DOI: 10.1007/978-1-60327-058-8_8. View

13.

Park J, Scheerer P, Hofmann K, Choe H, Ernst O . Crystal structure of the ligand-free G-protein-coupled receptor opsin. Nature. 2008; 454(7201):183-7. DOI: 10.1038/nature07063. View

14.

Byrne E, Sircar R, Miller P, Hedger G, Luchetti G, Nachtergaele S . Structural basis of Smoothened regulation by its extracellular domains. Nature. 2016; 535(7613):517-522. PMC: 4970916. DOI: 10.1038/nature18934. View

15.

Albert A, Boesze-Battaglia K, Paw Z, Watts A, Epand R . Effect of cholesterol on rhodopsin stability in disk membranes. Biochim Biophys Acta. 1996; 1297(1):77-82. DOI: 10.1016/0167-4838(96)00102-1. View

16.

Paila Y, Chattopadhyay A . The function of G-protein coupled receptors and membrane cholesterol: specific or general interaction?. Glycoconj J. 2008; 26(6):711-20. DOI: 10.1007/s10719-008-9218-5. View

17.

Castillo C, Albasanz J, Fernandez M, Martin M . Endogenous expression of adenosine A1, A2 and A3 receptors in rat C6 glioma cells. Neurochem Res. 2007; 32(6):1056-70. DOI: 10.1007/s11064-006-9273-x. View

18.

Sana T, Roark J, Li X, Waddell K, Fischer S . Molecular formula and METLIN Personal Metabolite Database matching applied to the identification of compounds generated by LC/TOF-MS. J Biomol Tech. 2009; 19(4):258-66. PMC: 2567134. View

19.

OMalley M, Helgeson M, Wagner N, Robinson A . The morphology and composition of cholesterol-rich micellar nanostructures determine transmembrane protein (GPCR) activity. Biophys J. 2011; 100(2):L11-3. PMC: 3021673. DOI: 10.1016/j.bpj.2010.12.3698. View

20.

Grest , KREMER . Molecular dynamics simulation for polymers in the presence of a heat bath. Phys Rev A Gen Phys. 1986; 33(5):3628-3631. DOI: 10.1103/physreva.33.3628. View