Guixà González, Ramon, 1978-Albasanz, José L.Rodriguez-Espigares, IsmaelPastor Maeso, ManuelSanz, FerranMartí Solano, MariaManna, MoutusiMartínez Seara, HectorHildebrand, Peter W.Martín, MairenaSelent, Jana2017-05-232017-05-232017Guixà González R, Albasanz JL, Rogriguez-Espigares I, Pastor Maeso M, Sanz F, Martí Solano M et al. Membrane cholesterol access into a G-protein-coupled receptor. Nature Communications. 2017;8:14505. DOI: 10.1038/ncomms145052041-1723http://hdl.handle.net/10230/32148Cholesterol 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 A2A receptor (A2AR) 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 A2AR-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 A2AR interior in a biotinylation assay. Overall, we show that cholesterol’s impact on A2AR-binding affinity goes beyond pure allosteric modulation and unveils a new interaction mode between cholesterol and the A2AR that could potentially apply to other GPCRs.application/pdfeng© Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/articlehttp://dx.doi.org/10.1038/ncomms14505G protein-coupled receptorsmembrane lipidsMolecular dynamicsinfo:eu-repo/semantics/openAccess