title: Tuning the allosteric regulation of artificial muscarinic and dopaminergic ligand-gated potassium channels by protein engineering of G protein-coupled receptors.
abstract: Ligand-gated ion channels enable intercellular transmission of action potential through synapses by transducing biochemical messengers into electrical signal. We designed artificial ligand-gated ion channels by coupling G protein-coupled receptors to the Kir6.2 potassium channel. These artificial channels called ion channel-coupled receptors offer complementary properties to natural channels by extending the repertoire of ligands to those recognized by the fused receptors, by generating more sustained signals and by conferring potassium selectivity. The first artificial channels based on the muscarinic M2 and the dopaminergic D2Lreceptors were opened and closed by acetylcholine and dopamine, respectively. We find here that this opposite regulation of the gating is linked to the length of the receptor C-termini, and that C-terminus engineering can precisely control the extent and direction of ligand gating. These findings establish the design rules to produce customized ligand-gated channels for synthetic biology applications.
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Christophe J
authors.initials: CJ
authors.lastname: Moreau
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Revilloud
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Lydia N
authors.initials: LN
authors.lastname: Caro
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Julien P
authors.initials: JP
authors.lastname: Dupuis
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Amandine
authors.initials: A
authors.lastname: Trouchet
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Argel
authors.initials: A
authors.lastname: Estrada-Mondragón
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Nieścierowicz
authors.affiliation: Laboratoire de Chimie et Biologie des Métaux (LCBM), CEA, Institut de Biosciences et Biotechnologies de Grenoble (BIG), UMR 5249, 38054 Grenoble, France.|LCBM, Univ. Grenoble Alpes, 38054 Grenoble, France.|LCBM, CNRS, 38054 Grenoble, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Sapay
authors.affiliation: Laboratoire de Chimie et Biologie des Métaux (LCBM), CEA, Institut de Biosciences et Biotechnologies de Grenoble (BIG), UMR 5249, 38054 Grenoble, France.|LCBM, Univ. Grenoble Alpes, 38054 Grenoble, France.|LCBM, CNRS, 38054 Grenoble, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Crouzy
authors.affiliation: Institut de Biologie Structurale (IBS), 38044 Grenoble, France.|IBS, Univ. Grenoble Alpes, 38044 Grenoble, France.|IBS, CNRS, LabEx ICST, 38044 Grenoble, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Vivaudou
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000494:Allosteric Regulation; D000818:Animals; D004298:Dopamine / Q000494:pharmacology; D058446:Ligand-Gated Ion Channels / Q000378:metabolism; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D043585:Receptor, Muscarinic M2 / Q000378:metabolism*; D017448:Receptors, Dopamine D2 / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014981:Xenopus; D000099161:Kcnj11 Channel
chemical_list: D000109:Acetylcholine; D004298:Dopamine; D058446:Ligand-Gated Ion Channels; D024661:Potassium Channels, Inwardly Rectifying; D043585:Receptor, Muscarinic M2; D017448:Receptors, Dopamine D2; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D000099161:Kcnj11 Channel; C113776:dopamine D2L receptor