title: A prokaryotic proton-gated ion channel from the nicotinic acetylcholine receptor family.
abstract: Ligand-gated ion channels (LGICs) mediate excitatory and inhibitory transmission in the nervous system. Among them, the pentameric or 'Cys-loop' receptors (pLGICs) compose a family that until recently was found in only eukaryotes. Yet a recent genome search identified putative homologues of these proteins in several bacterial species. Here we report the cloning, expression and functional identification of one of these putative homologues from the cyanobacterium Gloeobacter violaceus. It was expressed as a homo-oligomer in HEK 293 cells and Xenopus oocytes, generating a transmembrane cationic channel that is opened by extracellular protons and shows slow kinetics of activation, no desensitization and a single channel conductance of 8 pS. Electron microscopy and cross-linking experiments of the protein fused to the maltose-binding protein and expressed in Escherichia coli are consistent with a homo-pentameric organization. Sequence comparison shows that it possesses a compact structure, with the absence of the amino-terminal helix, the canonical disulphide bridge and the large cytoplasmic domain found in eukaryotic pLGICs. Therefore it embodies a minimal structure required for signal transduction. These data establish the prokaryotic origin of the family. Because Gloeobacter violaceus carries out photosynthesis and proton transport at the cytoplasmic membrane, this new proton-gated ion channel might contribute to adaptation to pH change.
authors.affiliation: Unit of Receptor and Cognition, CNRS URA D2182, Pasteur Institute, 75015 Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bocquet
authors.forename: Lia
authors.initials: L
authors.lastname: Prado de Carvalho
authors.forename: Jean
authors.initials: J
authors.lastname: Cartaud
authors.forename: Jacques
authors.initials: J
authors.lastname: Neyton
authors.forename: Chantal
authors.initials: C
authors.lastname: Le Poupon
authors.forename: Antoine
authors.initials: A
authors.lastname: Taly
authors.forename: Thomas
authors.initials: T
authors.lastname: Grutter
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Changeux
authors.forename: Pierre-Jean
authors.initials: PJ
authors.lastname: Corringer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D004553:Electric Conductivity; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011387:Prokaryotic Cells / Q000378:metabolism; D011487:Protein Conformation; D011522:Protons; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000145:classification*; D014981:Xenopus
chemical_list: D007473:Ion Channels; D011522:Protons; D011978:Receptors, Nicotinic