title: The biologically crucial C terminus of cholecystokinin and the non-peptide agonist SR-146,131 share a common binding site in the human CCK1 receptor. Evidence for a crucial role of Met-121 in the activation process.
abstract: The cholecystokinin (CCK) receptor-1 (CCK1R) is a G protein-coupled receptor, which mediates important central and peripheral cholecystokinin actions. Our aim was to progress in mapping of the CCK1R binding site by identifying residues that interact with the methionine and phenylalanine residues of the C-terminal moiety of CCK because these are crucial for its binding and biological activity, and to determine whether CCK and the selective non-peptide agonist, SR-146,131, share a common binding site. Identification of putative amino acids of the CCK1R binding site was achieved by dynamics-based docking of the ligand CCK in a refined three-dimensional model of the CCK1R using, as constraints, previous results that identified contact points between residues of CCK and CCK1R (Kennedy, K., Gigoux, V., Escrieut, C., Maigret, B., Martinez, J., Moroder, L., Frehel, D., Gully, D., Vaysse, N., and Fourmy, D. (1997) J. Biol. Chem. 272, 2920-2926 and Gigoux, V., Escrieut, C., Fehrentz, J. A., Poirot, S., Maigret, B., Moroder, L., Gully, D., Martinez, J., Vaysse, N., and Fourmy, D. (1999) J. Biol. Chem. 274, 20457-20464). By this approach, a series of residues forming connected hydrophobic clusters were identified. Pharmacological and functional analysis of mutated receptors indicated that a network of hydrophobic residues including Cys-94, Met-121, Val-125, Phe-218, Ile-329, Phe-330, Trp-326, Ile-352, Leu-356, and Tyr-360, is involved in the binding site for CCK and in the activation process of the CCK1R. Within this hydrophobic network, the physico-chemical nature of residue 121 seems to be essential for CCK1R functioning. Finally, the biological properties of mutants together with dynamic docking of SR-146,131 in the CCK1R binding site demonstrated that SR-146,131 occupies a region of CCK1R binding site which interacts with the C-terminal amidated tripeptide of CCK, i.e. Met-Asp-Phe-NH(2). These new and important insights will serve to better understand the activation process of CCK1R and to design or optimize ligands.
authors.affiliation: INSERM Unite 531, Institut Louis Bugnard, Centre Hospitalier Universitaire Rangueil, Bat. L3, 31403 Toulouse Cedex 4, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Escrieut
authors.forename: Véronique
authors.initials: V
authors.lastname: Gigoux
authors.forename: Elodie
authors.initials: E
authors.lastname: Archer
authors.forename: Sophie
authors.initials: S
authors.lastname: Verrier
authors.forename: Bernard
authors.initials: B
authors.lastname: Maigret
authors.forename: Raymond
authors.initials: R
authors.lastname: Behrendt
authors.forename: Luis
authors.initials: L
authors.lastname: Moroder
authors.forename: Eric
authors.initials: E
authors.lastname: Bignon
authors.forename: Sandrine
authors.initials: S
authors.lastname: Silvente-Poirot
authors.forename: Lucien
authors.initials: L
authors.lastname: Pradayrol
authors.forename: Daniel
authors.initials: D
authors.lastname: Fourmy
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002766:Cholecystokinin / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007211:Indoles / Q000737:chemistry*; D007700:Kinetics; D008024:Ligands; D008715:Methionine / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010649:Phenylalanine / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D043886:Receptor, Cholecystokinin A; D011949:Receptors, Cholecystokinin / Q000819:agonists / Q000737:chemistry* / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry*; D014162:Transfection
chemical_list: D000596:Amino Acids; D007211:Indoles; D008024:Ligands; D043886:Receptor, Cholecystokinin A; D011949:Receptors, Cholecystokinin; C118846:SR 146131; D013844:Thiazoles; D010649:Phenylalanine; D002766:Cholecystokinin; D008715:Methionine