Changes in amino-terminal portion of human B2 receptor selectively increase efficacy of synthetic ligand HOE 140 but not of cognate ligand bradykinin.
Recently, we have shown that a widely used antagonist of the human bradykinin B(2) receptor (B(2)R) HOE 140 acts as a full agonist of the chicken ornithokinin receptor (B(o)R). To understand the molecular mechanisms underlying differential efficacy of HOE 140 for the various kinin receptors, we have constructed chimeric kinin receptors (CKR) in which the amino-terminal portion including the first two transmembrane regions and the first extracellular loop (CKR-2) or only the second transmembrane region and the first extracellular loop (CKR-1) of B(2)R were substituted with the corresponding segments of B(o)R. Ligand efficacy of synthetic ligand HOE 140 decreased in the order B(o)R > CKR-2 > CKR-1 > B(2)R, whereas the efficacy of the endogenous kinin ligand was unchanged. Enhanced HOE 140 efficacy was not due to a structural change in the ligand binding site or to an enhanced receptor expression level. Rather, heterologous binding competition studies indicated that structural change(s) introduced into the engineered receptors caused a selective reduction in apparent affinity of HOE 140 for the uncoupled inactive receptor state R but not for the active G protein-coupled state R*, thereby increasing the ratio of R* over R for a given ligand concentration. Our results may help explain the unusually broad efficacy spectrum of HOE 140, which varies from inverse to full agonism, depending on kinin receptor subtype, tissue origin, or species.
Schroeder
Christian
C
Institute for Cardiovascular and Arteriosclerosis Research, Bayer D-42096, Wuppertal, Germany.
Breit
Andreas
A
Böning
Hilke
H
Dedio
Jürgen
J
Gera
Lajos
L
Stewart
John
J
Müller-Esterl
Werner
W
D019943:Amino Acid Substitution; D000818:Animals; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007295:Inositol Phosphates / Q000494:pharmacology; D008024:Ligands; D008968:Molecular Conformation; D043782:Receptor, Bradykinin B2; D018002:Receptors, Bradykinin / Q000096:biosynthesis / Q000737:chemistry* / Q000187:drug effects*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry
D018076:DNA, Complementary; D007295:Inositol Phosphates; D008024:Ligands; D043782:Receptor, Bradykinin B2; D018002:Receptors, Bradykinin; D011993:Recombinant Fusion Proteins; C065679:icatibant; D001920:Bradykinin