title: Structural implication for receptor oligomerization from functional reconstitution studies of mutant V2 vasopressin receptors.
abstract: Previous studies have established that G-protein-coupled receptors (GPCRs) are composed of independent folding domains. Based on this findings we attempted to rescue the function of clinically relevant missense mutations (R137H, S167L, and R181C) within the N-terminal domain of the V2 vasopressin receptor (V2-R), by coexpressing mutated full-length (Y280C) and C-terminally truncated (E242X) receptor constructs in COS-7 cells. Coimmunoprecipitation and enzyme-linked immunosorbent assay studies demonstrated a specific association of E242X with full-length V2-Rs even in the presence of missense mutations. Systematic analysis of the structural requirements for the observed receptor/fragment association showed that N-terminal fragments containing at least transmembrane regions 1-3 interact with the full-length V2-R. Despite this specific interaction, no functional reconstitution was achieved for mutant V2-Rs following coexpression with E242X and Y280C. However, functional activity of R137H and R181C upon coexpression with E242X was regained by mutational disruption of the extracellular disulfide bond, which is highly conserved among GPCRs. Our data with the V2-R are consistent with a structural model in which class I GPCRs form contact oligomers by lateral interaction rather than by a domain-swapping mechanism.
authors.affiliation: Institut für Pharmakologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, D-14195 Berlin, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Schulz
authors.forename: R
authors.initials: R
authors.lastname: Grosse
authors.forename: G
authors.initials: G
authors.lastname: Schultz
authors.forename: T
authors.initials: T
authors.lastname: Gudermann
authors.forename: T
authors.initials: T
authors.lastname: Schöneberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001704:Biopolymers; D019556:COS Cells; D004220:Disulfides / Q000737:chemistry; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017483:Receptors, Vasopressin / Q000737:chemistry* / Q000235:genetics
chemical_list: D001704:Biopolymers; D004220:Disulfides; D017483:Receptors, Vasopressin