title: Lysine 182 of endothelin B receptor modulates agonist selectivity and antagonist affinity: evidence for the overlap of peptide and non-peptide ligand binding sites.
abstract: The potent vasoactive peptide hormone endothelin (ET) binds to receptors which belong to the G-protein coupled receptor family. The availability of non-peptide antagonists for ET receptors allows investigation of the relationship among the binding sites for peptide and non-peptide ligands. In this study, a lysine residue, conserved within transmembrane domain 3 (TM3) of the ETA and ETB receptor subtypes, is implicated in agonist and antagonist binding by its analogous position within TM3 to a binding site aspartate residue conserved within bioactive amine receptors. Replacement of this lysine within hETB by arginine, alanine, methionine, aspartate, or glutamate results in hETB variants with unaltered affinities for agonist peptide ET-1 but which have affinities for peptide agonists ET-2, ET-3, sarafotoxin 6C, and TRL 1736 which are between 1-3 orders of magnitude lower than their corresponding wild-type hETB values. Significantly, the affinities of non-peptide antagonists, (+/-)-SB 209670 and its analogs as well as Ro 46-2005, are abrogated. The results suggest that an interaction of K182 of hETB with the indan 2-carboxyl of (+/-)-SB 209670 may contribute to the high-affinity binding of the diarylindan antagonists. The results indicate that TM3 of hETB is a region of overlap among the binding sites of non-peptide antagonists and the affected peptide agonists.
authors.affiliation: Department of Macromolecular Sciences, SmithKline Beechman Pharmaceuticals, King of Prussia, Pennsylvania 19406.
authors.forename: J A
authors.initials: JA
authors.lastname: Lee
authors.forename: J A
authors.initials: JA
authors.lastname: Brinkmann
authors.forename: E D
authors.initials: ED
authors.lastname: Longton
authors.forename: C E
authors.initials: CE
authors.lastname: Peishoff
authors.forename: M A
authors.initials: MA
authors.lastname: Lago
authors.forename: J D
authors.initials: JD
authors.lastname: Leber
authors.forename: R D
authors.initials: RD
authors.lastname: Cousins
authors.forename: A
authors.initials: A
authors.lastname: Gao
authors.forename: J M
authors.initials: JM
authors.lastname: Stadel
authors.forename: C S
authors.initials: CS
authors.lastname: Kumar
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D065128:Endothelin Receptor Antagonists; D016232:Endothelins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007189:Indans / Q000378:metabolism; D008024:Ligands; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D065128:Endothelin Receptor Antagonists; D016232:Endothelins; D007189:Indans; D008024:Ligands; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin; C087551:1H-Indene-2-carboxylic acid, 1-(1,3-benzodioxol-5-yl)-3-(2- (carboxymethoxy)-4-methoxyphenyl)-2,3-dihydro-5-propoxy-, (1S,2R,3S)-; D008239:Lysine