Altered agonist sensitivity of a mutant V2 receptor suggests a novel therapeutic strategy for nephrogenic diabetes insipidus

László Sándor Erdélyi, A. Balla, A. Patócs, Miklós Tóth, P. Várnai, L. Hunyady

Research output: Contribution to journalArticle

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Abstract

Loss-of-function mutations of the type 2 vasopressin receptor (V2R) in kidney can lead to nephrogenic diabetes insipidus (NDI). We studied a previously described, but uncharacterized, mutation of the V2R (N321K missense mutation) of a patient with NDI. The properties of the mutant receptor were evaluated. We constructed a highly sensitive Epac-based bioluminescence resonance energy transfer biosensor to perform real-time cAMP measurements after agonist stimulation of transiently transfected HEK293 cells with V2Rs. β-Arrestin binding of the activated receptors was examined with luciferase-tagged β-arrestin and mVenus-tagged V2Rs using the bioluminescence resonance energy transfer technique. Cell surface expression levels of hemagglutinin- tagged receptors were determined with flow cytometry using anti-hemagglutinin-Alexa 488 antibodies. Cellular localization examinations were implemented with fluorescent tagged receptors visualized with confocal laser scanning microscopy. The effect of various vasopressin analogs on the type 1 vasopressin receptor (V1R) was tested on mouse arteries by wire myography. The N321K mutant V2R showed normal cell surface expression, but the potency of arginine vasopressin for cAMP generation was low, whereas the clinically used desmopressin was not efficient. The β-arrestin binding and internalization properties of the mutant receptor were also different than those for the wild type. The function of the mutant receptor can be rescued with administration of the V2R agonist Val4-desmopressin, which had no detectable side effects on V1R in the effective cAMP generating concentrations. Based on these findings we propose a therapeutic strategy for patients with NDI carrying the N321K mutation, as our in vivo experiments suggest that Val4-desmopressin could rescue the function of the N321K-V2R without significant side effects on the V1R.

Original languageEnglish
Pages (from-to)634-643
Number of pages10
JournalMolecular Endocrinology
Volume28
Issue number5
DOIs
Publication statusPublished - 2014

Fingerprint

Nephrogenic Diabetes Insipidus
Arrestin
Deamino Arginine Vasopressin
Vasopressin Receptors
Hemagglutinins
Bioluminescence Resonance Energy Transfer Techniques
Mutation
Myography
Arginine Vasopressin
HEK293 Cells
Energy Transfer
Biosensing Techniques
Missense Mutation
Luciferases
Vasopressins
Confocal Microscopy
Flow Cytometry
Therapeutics
Arteries
Kidney

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology
  • Medicine(all)

Cite this

Altered agonist sensitivity of a mutant V2 receptor suggests a novel therapeutic strategy for nephrogenic diabetes insipidus. / Erdélyi, László Sándor; Balla, A.; Patócs, A.; Tóth, Miklós; Várnai, P.; Hunyady, L.

In: Molecular Endocrinology, Vol. 28, No. 5, 2014, p. 634-643.

Research output: Contribution to journalArticle

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