Pharmacological chaperones rescue cell-surface expression and function of misfolded V2 vasopressin receptor mutants
Jean-Pierre Morello, … , Daniel G. Bichet, Michel Bouvier
Jean-Pierre Morello, … , Daniel G. Bichet, Michel Bouvier
Published April 1, 2000
Citation Information: J Clin Invest. 2000;105(7):887-895. https://doi.org/10.1172/JCI8688.
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Article

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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Sensitivity of the mature and precursor V2R species to Endo H and PNGase...
Sensitivity of the mature and precursor V2R species to Endo H and PNGase F. The nature of the del 62-64 V2R species was assessed in pulse-chase metabolic-labeling experiments carried out in cells treated (or not) with SR121463A. The glycosylation state was determined at the 60-minute chase time by treating the purified receptor with the indicated enzymes. In the absence of SR121463A treatment (a), only the precursor (open arrowhead) form that is sensitive to both enzymes is observed, whereas following a 10–5-M SR121463A treatment for 16 hours (b), the Endo H–resistant mature species is observed (closed arrowhead). (c) Concentration dependence of the SR121463A-promoted maturation of the V2R. The effect of the indicated concentrations of SR121463A (for 16 hours) on the maturation of the del 62-64 V2R was assessed at the 60–minute chase time. (d) The mature/precursor receptor ratio of the autoradiogram shown in c was calculated by densitometric analysis of the autoradiogram. Representative of 3 separate experiments.