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

• Published in: Molecular pharmacology Vol. 53; no. 4; p. 766
• Main Authors: Porter, J E, Edelmann, S E, Waugh, D J, Piascik, M T, Perez, D M
• Format: Journal Article
• Language: English
• Published: United States 01-04-1998
• Subjects: Adrenergic alpha-1 Receptor Agonists; Animals; Binding Sites - genetics; Cell Line; COS Cells; Cricetinae; Drug Synergism; Ethylamines - metabolism; Ethylamines - pharmacology; Humans; Inositol Phosphates - metabolism; Mutagenesis, Site-Directed; Protein Structure, Secondary; Rats; Receptors, Adrenergic, alpha-1 - chemistry; Receptors, Adrenergic, alpha-1 - genetics; Receptors, Adrenergic, alpha-1 - metabolism; Sodium Chloride - chemistry; Sodium Chloride - pharmacology; Solubility; Structure-Activity Relationship
• ISSN: 0026-895X
• DOI: 10.1124/mol.53.4.766

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.