Grimes, JakKoszegi, ZsomborLanoiselée, YannMiljus, TamaraO'Brien, Shannon L.Stepniewski, Tomasz Maciej, 1988-Medel Lacruz, BrianBaidya, MithuMakarova, MariaMistry, RaviGoulding, JoëlleDrube, JuliaHoffmann, CarstenOwen, Dylan M.Shukla, Arun K.Selent, JanaHill, Stephen J.Calebiro, Davide2023-09-082023-09-082023Grimes J, Koszegi Z, Lanoiselée Y, Miljus T, O’Brien SL, Stepniewski TM, Medel-Lacruz B, Baidya M, Makarova M, Mistry R, Goulding J, Drube J, Hoffmann C, Owen DM, Shukla AK, Selent J, Hill SJ. Plasma membrane preassociation drives β-arrestin coupling to receptors and activation. Cell. 2023;186(10):2238-55. DOI: 10.1016/j.cell.2023.04.0180092-8674http://hdl.handle.net/10230/57839β-arrestin plays a key role in G protein-coupled receptor (GPCR) signaling and desensitization. Despite recent structural advances, the mechanisms that govern receptor-β-arrestin interactions at the plasma membrane of living cells remain elusive. Here, we combine single-molecule microscopy with molecular dynamics simulations to dissect the complex sequence of events involved in β-arrestin interactions with both receptors and the lipid bilayer. Unexpectedly, our results reveal that β-arrestin spontaneously inserts into the lipid bilayer and transiently interacts with receptors via lateral diffusion on the plasma membrane. Moreover, they indicate that, following receptor interaction, the plasma membrane stabilizes β-arrestin in a longer-lived, membrane-bound state, allowing it to diffuse to clathrin-coated pits separately from the activating receptor. These results expand our current understanding of β-arrestin function at the plasma membrane, revealing a critical role for β-arrestin preassociation with the lipid bilayer in facilitating its interactions with receptors and subsequent activation.application/pdfeng© 2023 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).info:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.cell.2023.04.018G protein-coupled receptorsGPCRTIRFArrestinPlasma membraneProtein-protein interactionsSingle-molecule microscopyinfo:eu-repo/semantics/openAccess