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Titolo:
Exploration of the ligand binding site of the human 5-HT4 receptor by site-directed mutagenesis and molecular modeling
Autore:
Mialet, J; Dahmoune, Y; Lezoualch, F; Berque-Bestel, I; Eftekhari, P; Hoebeke, J; Sicsic, S; Langlois, M; Fischmeister, R;
Indirizzi:
Univ Paris Sud, Fac Pharm, ISIT, IFR,UPRES A 8076, F-92296 Chatenay Malabry, France Univ Paris Sud Chatenay Malabry France F-92296 Chatenay Malabry, France Univ Paris Sud, Fac Pharm, INSERM U 446, Lab Cardiol Cellulaire & Mol, F-92296 Chatenay Malabry, France Univ Paris Sud Chatenay Malabry France F-92296 Chatenay Malabry, France Univ Paris Sud, Fac Pharm, UPRES A CNRS 8076, Lab Reconnaissance Mol & Cellulaire BIOCIS, F-92296 Chatenay Malabry, France Univ Paris Sud Chatenay Malabry France F-92296 Chatenay Malabry, France CNRS, UPR 9021, F-67084 Strasbourg, France CNRS Strasbourg France F-67084 NRS, UPR 9021, F-67084 Strasbourg, France
Titolo Testata:
BRITISH JOURNAL OF PHARMACOLOGY
fascicolo: 3, volume: 130, anno: 2000,
pagine: 527 - 538
SICI:
0007-1188(200006)130:3<527:EOTLBS>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-COUPLED RECEPTORS; 3-DIMENSIONAL MODELS; AGONIST BINDING; AMINO-ACIDS; SEROTONIN; 5-HYDROXYTRYPTAMINE(2A); IDENTIFICATION; PHARMACOLOGY; ACTIVATION; SUBTYPES;
Keywords:
serotonin; 5-HT4 receptor; human; binding site; directed mutagenesis; molecular modeling;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Sicsic, S Univ Paris Sud, Fac Pharm, ISIT, IFR,UPRES A 8076, 5 Rue JB Clement, F-92296 Chatenay Malabry, France Univ Paris Sud 5 Rue JB Clement Chatenay Malabry France F-92296
Citazione:
J. Mialet et al., "Exploration of the ligand binding site of the human 5-HT4 receptor by site-directed mutagenesis and molecular modeling", BR J PHARM, 130(3), 2000, pp. 527-538

Abstract

1 Among the five human 5-HT4 (h5-HT4) receptor isoforms, the hS-HT4(a) receptor was studied with a particular emphasis on the molecular interactions involved in ligand binding. For this purpose, we used site-directed mutagenesis of the transmembrane domain. Twelve mutants were constructed with a special focus on the residue P4.53 of helix IV which substitutes in h5-HT4 receptors the highly conserved S residue among the rhodopsin family receptors. The mutated receptors were transiently expressed in COS-7 cells.2 Ligand binding or competition studies with two h5-HT4 receptor agonists,serotonin and ML10302 and two h5-HT4 receptor antagonists, [H-3]-GR113808 and ML10375 were performed on wild type and mutant receptors. Functional activity of the receptors was evaluated by measuring the ability of serotoninto stimulate adenylyl cyclase.3 Ligand binding experiments revealed that [H-3]-GR113808 did not bind to mutants P4.53A, S5.43A, F6.51A, Y7.43A and to double mutant F6.52V/N6.55L. On the other hand mutations D3.32N, S5.43A and Y7.43A appeared to promote adramatic decrease of h5-HT4(a) receptor functional activity. From these studies, S5.43 and Y7.43 clearly emerged as common anchoring sites to antagonist [H-3]-GR113808 and to serotonin.4 According to these results, we propose ligand-receptor complex models with serotonin and [H-3]-GR113808. For serotonin, three interaction points were selected including ionic interaction with D3.32, a stabilizing interaction of this ion pair by Y7.43 and a hydrogen bond with S5.43. [H-3]-GR113808was also docked, based on the same type of interactions with S5.43 and D3.32: the proposed model suggested a possible role of P4.53 in helix IV structure allowing the involvement of a close hydrophobic residue, W4.50, in a hydrophobic pocket for hydrophobic interactions with the indole ring of [H-3]-GR113808.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 25/11/20 alle ore 07:33:47