title: Water-soluble nicotinic acetylcholine receptor formed by alpha7 subunit extracellular domains.
abstract: Water-soluble models of ligand-gated ion channels would be advantageous for structural studies. We investigated the suitability of three versions of the N-terminal extracellular domain (ECD) of the alpha7 subunit of the nicotinic acetylcholine receptor (AChR) family for this purpose by examining their ligand-binding and assembly properties. Two versions included the first transmembrane domain and were solubilized with detergent after expression in Xenopus oocytes. The third was truncated before the first transmembrane domain and was soluble without detergent. For all three, their equilibrium binding affinities for alpha-bungarotoxin, nicotine, and acetylcholine, combined with their velocity sedimentation profiles, were consistent with the formation of native-like AChRs. These characteristics imply that the alpha7 ECD can form a water-soluble AChR that is a model of the ECD of the full-length alpha7 AChR.
authors.affiliation: Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6082, USA. wellsg@mail.med.upenn.edu
authors.forename: G B
authors.initials: GB
authors.lastname: Wells
authors.forename: R
authors.initials: R
authors.lastname: Anand
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: J
authors.initials: J
authors.lastname: Lindstrom
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D002645:Chickens; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D009538:Nicotine / Q000378:metabolism; D011485:Protein Binding; D011869:Radioligand Assay; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D014867:Water
chemical_list: D002038:Bungarotoxins; D007457:Iodine Radioisotopes; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; D014867:Water; D009538:Nicotine; D000109:Acetylcholine