A Molecular Mechanism for Sequential Activation of a G Protein-Coupled Receptor

TY - JOUR

T1 - A Molecular Mechanism for Sequential Activation of a G Protein-Coupled Receptor

AU - Grundmann, Manuel

AU - Tikhonova, Irina G

AU - Hudson, Brian D

AU - Smith, Nicola J

AU - Mohr, Klaus

AU - Ulven, Trond

AU - Milligan, Graeme

AU - Kenakin, Terry

AU - Kostenis, Evi

N1 - Copyright © 2016 Elsevier Ltd. All rights reserved.

PY - 2016/3/17

Y1 - 2016/3/17

N2 - Ligands targeting G protein-coupled receptors (GPCRs) are currently classified as either orthosteric, allosteric, or dualsteric/bitopic. Here, we introduce a new pharmacological concept for GPCR functional modulation: sequential receptor activation. A hallmark feature of this is a stepwise ligand binding mode with transient activation of a first receptor site followed by sustained activation of a second topographically distinct site. We identify 4-CMTB (2-(4-chlorophenyl)-3-methyl-N-(thiazol-2-yl)butanamide), previously classified as a pure allosteric agonist of the free fatty acid receptor 2, as the first sequential activator and corroborate its two-step activation in living cells by tracking integrated responses with innovative label-free biosensors that visualize multiple signaling inputs in real time. We validate this unique pharmacology with traditional cellular readouts, including mutational and pharmacological perturbations along with computational methods, and propose a kinetic model applicable to the analysis of sequential receptor activation. We envision this form of dynamic agonism as a common principle of nature to spatiotemporally encode cellular information.

AB - Ligands targeting G protein-coupled receptors (GPCRs) are currently classified as either orthosteric, allosteric, or dualsteric/bitopic. Here, we introduce a new pharmacological concept for GPCR functional modulation: sequential receptor activation. A hallmark feature of this is a stepwise ligand binding mode with transient activation of a first receptor site followed by sustained activation of a second topographically distinct site. We identify 4-CMTB (2-(4-chlorophenyl)-3-methyl-N-(thiazol-2-yl)butanamide), previously classified as a pure allosteric agonist of the free fatty acid receptor 2, as the first sequential activator and corroborate its two-step activation in living cells by tracking integrated responses with innovative label-free biosensors that visualize multiple signaling inputs in real time. We validate this unique pharmacology with traditional cellular readouts, including mutational and pharmacological perturbations along with computational methods, and propose a kinetic model applicable to the analysis of sequential receptor activation. We envision this form of dynamic agonism as a common principle of nature to spatiotemporally encode cellular information.

U2 - 10.1016/j.chembiol.2016.02.014

DO - 10.1016/j.chembiol.2016.02.014

M3 - Journal article

SN - 2451-9448

VL - 23

SP - 392

EP - 403

JO - Chemistry and Biology

JF - Chemistry and Biology

IS - 3

ER -