title: Mechanism of activation of a G protein-coupled receptor, the human cholecystokinin-2 receptor.
abstract: G protein-coupled receptors (GPCRs) represent a major focus in functional genomics programs and drug development research, but their important potential as drug targets contrasts with the still limited data available concerning their activation mechanism. Here, we investigated the activation mechanism of the cholecystokinin-2 receptor (CCK2R). The three-dimensional structure of inactive CCK2R was homology-modeled on the basis of crystal coordinates of inactive rhodopsin. Starting from the inactive CCK2R modeled structure, active CCK2R (namely cholecystokinin-occupied CCK2R) was modeled by means of steered molecular dynamics in a lipid bilayer and by using available data from other GPCRs, including rhodopsin. By comparing the modeled structures of the inactive and active CCK2R, we identified changes in the relative position of helices and networks of interacting residues, which were expected to stabilize either the active or inactive states of CCK2R. Using targeted molecular dynamics simulations capable of converting CCK2R from the inactive to the active state, we delineated structural changes at the atomic level. The activation mechanism involved significant movements of helices VI and V, a slight movement of helices IV and VII, and changes in the position of critical residues within or near the binding site. The mutation of key amino acids yielded inactive or constitutively active CCK2R mutants, supporting this proposed mechanism. Such progress in the refinement of the CCK2R binding site structure and in knowledge of CCK2R activation mechanisms will enable target-based optimization of nonpeptide ligands.
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France.
authors.forename: Esther
authors.initials: E
authors.lastname: Marco
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Foucaud
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Langer
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Escrieut
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France.
authors.forename: Irina G
authors.initials: IG
authors.lastname: Tikhonova
authors.affiliation: INSERM, Unit 858, Institut Fédératif de Recherche, 31432 Toulouse, France and the Université de Toulouse 3, 31062 Toulouse, France; Université de Toulouse 3, 31062 Toulouse, France. Electronic address: fourmyd@toulouse.inserm.fr.
authors.forename: Daniel
authors.initials: D
authors.lastname: Fourmy
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D002766:Cholecystokinin; D003198:Computer Simulation; D015195:Drug Design; D023281:Genomics; D006801:Humans; D008051:Lipid Bilayers; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D043887:Receptor, Cholecystokinin B / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012243:Rhodopsin; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship
chemical_list: D008051:Lipid Bilayers; D043887:Receptor, Cholecystokinin B; D012243:Rhodopsin; D002766:Cholecystokinin