title: Role of key aromatic residues in the ligand-binding domain of alpha7 nicotinic receptors in the agonist action of beta-amyloid.
abstract: Soluble β-amyloid (Aβ) resides in certain regions of the brain at or near picomolar concentration, rising in level during the prodromic stage of Alzheimer disease. Recently, we identified the homomeric α7 nicotinic acetylcholine receptor (α7-nAChR) as one possible functional target for picomolar Aβ. This study was aimed at addressing which residues in α7-nAChRs potentially interact with Aβ to regulate the presynaptic function of this receptor. Site-directed mutagenesis was carried out to study the key aromatic residues in the mouse α7-nAChR agonist-binding pocket. Mutations of tyrosine188 resulted in a decrease in activation of presynaptic α7-nAChRs by ACh and Aβ but with no change in response to nicotine, indicating the critical role of Tyr-188 in presynaptic regulation by Aβ. Coimmunoprecipitation additionally revealed direct binding of Aβ to α7-nAChRs and to the Tyr-188 mutant receptor. In contrast, mutations of Tyr-195 in α7-nAChR led to decreased activation by nicotine without apparent effects on ACh- or Aβ-induced responses. Agonist-induced responses of Tyr-93 mutant α7-nAChRs indicated possible interactions of nicotine and Aβ with its hydroxyl group, but there was no change in presynaptic responses after mutation of Trp-149. All of the mutants were shown to be expressed on the plasma membrane using cell surface labeling. Together, these results directly demonstrate an essential role for the aromatic residue Tyr-188 as a key component in the agonist binding domain for the activation of α7-nAChRs by Aβ.
authors.affiliation: Department of Cell and Molecular Biology, University of Hawai'i at Manoa, Honolulu, HI 96813, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Tong
authors.forename: Komal
authors.initials: K
authors.lastname: Arora
authors.forename: Michael M
authors.initials: MM
authors.lastname: White
authors.forename: Robert A
authors.initials: RA
authors.lastname: Nichols
mesh_terms: D024322:Amino Acids, Aromatic / Q000235:genetics / Q000378:metabolism*; D000682:Amyloid / Q000235:genetics / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D051379:Mice; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D010449:Peptide Mapping; D017434:Protein Structure, Tertiary; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism*; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: D024322:Amino Acids, Aromatic; D000682:Amyloid; D016564:Amyloid beta-Protein Precursor; C579166:Chrna7 protein, mouse; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor