title: The agonism and synergistic potentiation of weak partial agonists by triethylamine in alpha 1-adrenergic receptor activation: evidence for a salt bridge as the initiating process.
abstract: Alpha 1-adrenergic receptor (AR) activation is thought to be initiated by disruption of a constraining interhelical salt bridge (). Disruption of this salt bridge is achieved through a competition for the aspartic acid residue in transmembrane domain three by the protonated amine of the endogenous ligand norepinephrine and a lysine residue in transmembrane domain seven. To further test this hypothesis, we investigated the possibility that a simple amine could mimic an important functional group of the endogenous ligand and break this alpha 1-AR ionic constraint leading to agonism. Triethylamine (TEA) was able to generate concentration-dependent increases of soluble inositol phosphates in COS-1 cells transiently transfected with the hamster alpha 1b-AR and in Rat-1 fibroblasts stably transfected with the human alpha 1a-AR subtype. TEA was also able to synergistically potentiate the second messenger production by weak partial alpha 1-AR agonists and this effect was fully inhibited by the alpha 1-AR antagonist prazosin. However, this synergistic potentiation was not observed for full alpha 1-AR agonists. Instead, TEA caused a parallel rightward shift of the dose-response curve, consistent with the properties of competitive antagonism. TEA specifically bound to a single population of alpha 1-ARs with a Ki of 28.7 +/- 4.7 mM. In addition, the site of binding by TEA to the alpha 1-AR is at the conserved aspartic acid residue in transmembrane domain three, which is part of the constraining salt bridge. These results indicate a direct interaction of TEA in the receptor agonist binding pocket that leads to a disruption of the constraining salt bridge, thereby initiating alpha 1-AR activation.
authors.affiliation: Department of Molecular Cardiology, The Lerner Research Institute, The Cleveland Clinic Foundation, Ohio 44195, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Porter
authors.forename: S E
authors.initials: SE
authors.lastname: Edelmann
authors.forename: D J
authors.initials: DJ
authors.lastname: Waugh
authors.forename: M T
authors.initials: MT
authors.lastname: Piascik
authors.forename: D M
authors.initials: DM
authors.lastname: Perez
mesh_terms: D058646:Adrenergic alpha-1 Receptor Agonists; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D019556:COS Cells; D002460:Cell Line; D006224:Cricetinae; D004357:Drug Synergism; D005021:Ethylamines / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D051381:Rats; D018340:Receptors, Adrenergic, alpha-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012965:Sodium Chloride / Q000737:chemistry* / Q000494:pharmacology; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: C547713:ADRA1A protein, human; C547716:ADRA1B protein, human; C547715:Adra1a protein, rat; C547718:Adra1b protein, rat; D058646:Adrenergic alpha-1 Receptor Agonists; D005021:Ethylamines; D007295:Inositol Phosphates; D018340:Receptors, Adrenergic, alpha-1; D012965:Sodium Chloride; C016162:triethylamine