title: Functional characterization of five V2 vasopressin receptor gene mutations.
abstract: COS7 cells were transiently transfected with plasmids encoding mutant forms of the V2 vasopressin receptors corresponding to mutations [Y280C, L292P, R337stop, V277A, and G12E (the latter found in the same kindred with L292P)] recently identified in subjects with X-linked nephrogenic diabetes insipidus (NDI). cAMP response to dDAVP and AVP, saturation binding experiments with [3H]-AVP, immunofluorescence, and indirect ELISA studies were performed to characterize the functional consequences of these mutations. The Y280C, L292P, and R337stop mutant V2 receptors show substantially decreased cell surface expression and are functionally inactive. The V277A mutant receptor, though well expressed at the cell surface as seen by immunofluorescence and ELISA and having a dissociation constant with AVP similar to the wild type receptor, was functionally less active as seen by a substantially decreased receptor number (Bmax) and reduced cAMP stimulation by dDAVP. The G12E mutant was functionally the same as the wild type V2 receptor in both cAMP stimulation and binding. These results provide insight into residues critical for V2 receptor expression and function and also provide direct evidence that Y280C, L292P, R337stop and V277A mutations are the cause of X-linked NDI in affected subjects.
authors.affiliation: Metabolic Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: D
authors.initials: D
authors.lastname: Wenkert
authors.forename: T
authors.initials: T
authors.lastname: Schoneberg
authors.forename: J J
authors.initials: JJ
authors.lastname: Merendino
authors.forename: M S
authors.initials: MS
authors.lastname: Rodriguez Pena
authors.forename: R
authors.initials: R
authors.lastname: Vinitsky
authors.forename: P K
authors.initials: PK
authors.lastname: Goldsmith
authors.forename: J
authors.initials: J
authors.lastname: Wess
authors.forename: A M
authors.initials: AM
authors.lastname: Spiegel
mesh_terms: D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism / Q000494:pharmacology*; D019556:COS Cells; D002462:Cell Membrane / Q000737:chemistry; D000242:Cyclic AMP / Q000096:biosynthesis; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology; D018500:Diabetes Insipidus, Nephrogenic / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D009154:Mutation / Q000502:physiology*; D017483:Receptors, Vasopressin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins
chemical_list: D017483:Receptors, Vasopressin; D011993:Recombinant Fusion Proteins; D001127:Arginine Vasopressin; D000242:Cyclic AMP; D003894:Deamino Arginine Vasopressin