Pharmacological chaperones rescue cell-surface expression and function of misfolded V2 vasopressin receptor mutants
Jean-Pierre Morello, Ali Salahpour, André Laperrière, Virginie Bernier, Marie-Françoise Arthus, Michèle Lonergan, Ulla Petäjä-Repo, Stéphane Angers, Denis Morin, Daniel G. Bichet, Michel Bouvier
Jean-Pierre Morello, Ali Salahpour, André Laperrière, Virginie Bernier, Marie-Françoise Arthus, Michèle Lonergan, Ulla Petäjä-Repo, Stéphane Angers, Denis Morin, Daniel G. Bichet, Michel Bouvier
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Pharmacological chaperones rescue cell-surface expression and function of misfolded V2 vasopressin receptor mutants

Abstract

Over 150 mutations within the coding sequence of the V2 vasopressin receptor (V2R) gene are known to cause nephrogenic diabetes insipidus (NDI). A large number of these mutant receptors fail to fold properly and therefore are not routed to the cell surface. Here we show that selective, nonpeptidic V2R antagonists dramatically increase cell-surface expression and rescue the function of 8 mutant NDI-V2Rs by promoting their proper folding and maturation. A cell-impermeant V2R antagonist could not mimic these effects and was unable to block the rescue mediated by a permeant agent, indicating that the nonpeptidic antagonists act intracellularly, presumably by binding to and stabilizing partially folded mutants. In addition to opening new therapeutic avenues for NDI patients, these data demonstrate that by binding to newly synthesized mutant receptors, small ligands can act as pharmacological chaperones, promoting the proper folding and maturation of receptors and their targeting to the cell surface.

Authors

Jean-Pierre Morello, Ali Salahpour, André Laperrière, Virginie Bernier, Marie-Françoise Arthus, Michèle Lonergan, Ulla Petäjä-Repo, Stéphane Angers, Denis Morin, Daniel G. Bichet, Michel Bouvier

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Concentration-dependent increase of cell-surface del 62-64 V2R after SR1...
Concentration-dependent increase of cell-surface del 62-64 V2R after SR121463A treatment. Cell-surface receptor expression was measured by FACS of HEK-293 cells stably expressing either del 62-64 (a) or WT (b) V2R. Cells were treated with the indicated concentrations of SR121463A. Representative of 3 separate experiments. (c) Dose-dependent effect of SR121463A treatment on cell-surface del 62-64 (filled triangles) and WT (filled squares) V2R expression as quantified by the mean cell-surface fluorescence intensity obtained by FACS. The 100% point was taken from cells exposed to a 10–5 M concentration of SR121463A, and the 0% point was taken from cells incubated without anti-myc antibody. There was no difference in cell-surface fluorescence intensity between untransfected HEK-293 cells incubated with both antibodies and HEK-293 cells expressing the del 62-64 V2R that were incubated without anti-myc antibody. There was no difference in cell-surface fluorescence intensity between untransfected HEK-293 cells incubated with both antibodies and HEK-293 cells expressing the del 62-64 V2R that were incubated without anti-myc antibody. The data are expressed as means ± SEM (n = 3).