title: The effect of eight V2 vasopressin receptor mutations on stimulation of adenylyl cyclase and binding to vasopressin.
abstract: We previously identified six V2 vasopressin receptor mutations in five unrelated nephrogenic diabetes insipidus (NDI) families. In order to elucidate the effect of these mutations on the function of the V2 vasopressin receptor, we introduced these six and two additional, naturally occurring mutations into the V2 vasopressin receptor gene by in vitro mutagenesis. Five of the mutants (two frameshift, one nonsense, and two missense) failed to stimulate adenylyl cyclase due to their inability to bind vasopressin under the experimental conditions. In contrast, ligand binding and cAMP accumulation were normal for two other mutations, a A61V missense mutation and an in-frame deletion of four amino acids (Arg-247 to Gly-250), suggesting that they are not the cause of NDI in these families. The deletion mutation was found in a family in conjunction with a second mutation, R181C, which yielded a much reduced ligand-binding capacity. The KD of R181C was at least 26 times higher than that of the wild type. Further characterization by an immunofluorescent assay showed that the R181C mutant receptor is expressed and distributed on the cell surface in a manner similar to that of the wild type. This finding indicates that the inability of this mutant to stimulate adenylyl cyclase is caused by the reduced capacity for vasopressin binding and that the R181C mutation is responsible for NDI in this family.
authors.affiliation: Howard Hughes Medical Institute, University of California, San Francisco 94143.
authors.forename: Y
authors.initials: Y
authors.lastname: Pan
authors.forename: P
authors.initials: P
authors.lastname: Wilson
authors.forename: J
authors.initials: J
authors.lastname: Gitschier
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D000595:Amino Acid Sequence; D003919:Diabetes Insipidus / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009154:Mutation; D017483:Receptors, Vasopressin / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D014667:Vasopressins / Q000378:metabolism*
chemical_list: D000939:Epitopes; D017483:Receptors, Vasopressin; D014667:Vasopressins; D000262:Adenylyl Cyclases