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Titolo:
Arginine 197 of the cholecystokinin-A receptor binding site interacts withthe sulfate of the peptide agonist cholecystokinin
Autore:
Gigoux, V; Maigret, B; Escrieut, C; Silvente-Poirot, S; Bouisson, M; Fehrentz, JA; Moroder, L; Gully, D; Martinez, J; Vaysse, N; Fourmy, D;
Indirizzi:
CHU Rangueil, INSERM, U151, F-31054 Toulouse, France CHU Rangueil Toulouse France F-31054 ERM, U151, F-31054 Toulouse, France Univ Nancy, Chim Theor Lab, F-54506 Vandoeuvre Nancy, France Univ Nancy Vandoeuvre Nancy France F-54506 4506 Vandoeuvre Nancy, France Fac Pharm Montpellier, CNRS, UMR 5810, F-34060 Montpellier, France Fac Pharm Montpellier Montpellier France F-34060 060 Montpellier, France Max Planck Inst Biochem, D-82143 Martinsried, Germany Max Planck Inst Biochem Martinsried Germany D-82143 Martinsried, Germany Sanofi Rech, F-31036 Toulouse, France Sanofi Rech Toulouse France F-31036 anofi Rech, F-31036 Toulouse, France
Titolo Testata:
PROTEIN SCIENCE
fascicolo: 11, volume: 8, anno: 1999,
pagine: 2347 - 2354
SICI:
0961-8368(199911)8:11<2347:A1OTCR>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-COUPLED RECEPTORS; FUNCTIONAL EXPRESSION; MOLECULAR-CLONING; PANCREAS; IDENTIFICATION; GASTRIN; STATES; ACIDS;
Keywords:
cholecystokinin; modeling; mutagenesis; receptor; site; sulfate;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Fourmy, D CHU Rangueil, INSERM, U151, Bat L3, F-31054 Toulouse, France CHURangueil Bat L3 Toulouse France F-31054 54 Toulouse, France
Citazione:
V. Gigoux et al., "Arginine 197 of the cholecystokinin-A receptor binding site interacts withthe sulfate of the peptide agonist cholecystokinin", PROTEIN SCI, 8(11), 1999, pp. 2347-2354

Abstract

The knowledge of the binding sites of G protein-coupled cholecystokinin receptors represents important insights that may serve to understand their activation processes and to design or optimize ligands. Our aim was to identify the amino acid of the cholecystokinin-A receptor (CCK-AR) binding site in an interaction with the sulfate of CCK, which is crucial for CCK binding and activity. A three-dimensional model of the [CCK-AR-CCK] complex was built. In this model, Arg197 was the best candidate residue for a ionic interaction with the sulfate of CCK, Arg197 was exchanged for a methionine by site-directed mutagenesis. Wild-type and mutated CCK-AR were transiently expressed in COS-7 cells for pharmacological and functional analysis. The mutated receptor on Arg197 did not bind the agonist radioligand I-125- BH-[Thr, Nle]-CCK-9: however, it bound the nonpeptide antagonist [H-3]-SR27,897 as the wild-type receptor. The mutant was congruent to 1,470- and 3,200-fold less potent than the wild-type CCK-AR to activate G proteins and to induce inositol phosphate production, respectively. This is consistent with the 500-fold lower potency and 800-fold lower affinity of nonsulfated CCK relative to sulfated CCK on the wild-type receptor. These data, together with those showing that the mutated receptor failed to discriminate nonsulfated and sulfated CCK while it retained other pharmacological features of the CCK-ARI strongly support an interaction between Arg197 of the CCK-AR binding site and the sulfate of CCK. In addition, the mutated CCK-AR resembled the low affinity state of the wild-type CCK-AR, suggesting that Arg197-sulfate interaction regulates conformational changes of the CCK-AR that are required for its physiological activation.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 06:30:19