title: Nicotinic receptor binding site probed with unnatural amino acid incorporation in intact cells.
abstract: The nonsense codon suppression method for unnatural amino acid incorporation has been applied to intact cells and combined with electrophysiological analysis to probe structure-function relations in the nicotinic acetylcholine receptor. Functional receptors were expressed in Xenopus oocytes when tyrosine and phenylalanine derivatives were incorporated at positions 93, 190, and 198 in the binding site of the alpha subunit. Subtle changes in the structure of an individual side chain produced readily detectable changes in the function of this large channel protein. At each position, distinct features of side chain structure dominated the dose-response relation, probably by governing the agonist-receptor binding.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena 91125, USA.
authors.forename: M W
authors.initials: MW
authors.lastname: Nowak
authors.forename: P C
authors.initials: PC
authors.lastname: Kearney
authors.forename: J R
authors.initials: JR
authors.lastname: Sampson
authors.forename: M E
authors.initials: ME
authors.lastname: Saks
authors.forename: C G
authors.initials: CG
authors.lastname: Labarca
authors.forename: S K
authors.initials: SK
authors.lastname: Silverman
authors.forename: W
authors.initials: W
authors.lastname: Zhong
authors.forename: J
authors.initials: J
authors.lastname: Thorson
authors.forename: J N
authors.initials: JN
authors.lastname: Abelson
authors.forename: N
authors.initials: N
authors.lastname: Davidson
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003062:Codon; D006860:Hydrogen Bonding; D008024:Ligands; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry*; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry*; D014981:Xenopus
chemical_list: D003062:Codon; D008024:Ligands; D011978:Receptors, Nicotinic; D014443:Tyrosine; D010649:Phenylalanine