Functional characterization of five V2 vasopressin receptor gene mutations

David Wenkert, Torsten Schoneberg, John J. Merendino, Maria Sol Rodriguez Pena, Ruth Vinitsky, Paul K. Goldsmith, Jurgen Wess, Allen M. Spiegel

Research output: Contribution to journalArticle

57 Citations (Scopus)

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 (B(max)) 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.

Original languageEnglish (US)
Pages (from-to)43-50
Number of pages8
JournalMolecular and Cellular Endocrinology
Volume124
Issue number1-2
DOIs
StatePublished - Nov 29 1996
Externally publishedYes

Fingerprint

Vasopressin Receptors
Genes
Nephrogenic Diabetes Insipidus
Mutation
Medical problems
Enzyme-Linked Immunosorbent Assay
Indirect Fluorescent Antibody Technique
Fluorescent Antibody Technique
Plasmids
Experiments

Keywords

  • Expression
  • Mutations
  • Nephrogenic diabetes insipidus
  • Receptor
  • Transfection
  • Vasopressin

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Wenkert, D., Schoneberg, T., Merendino, J. J., Rodriguez Pena, M. S., Vinitsky, R., Goldsmith, P. K., ... Spiegel, A. M. (1996). Functional characterization of five V2 vasopressin receptor gene mutations. Molecular and Cellular Endocrinology, 124(1-2), 43-50. https://doi.org/10.1016/S0303-7207(96)03926-3

Functional characterization of five V2 vasopressin receptor gene mutations. / Wenkert, David; Schoneberg, Torsten; Merendino, John J.; Rodriguez Pena, Maria Sol; Vinitsky, Ruth; Goldsmith, Paul K.; Wess, Jurgen; Spiegel, Allen M.

In: Molecular and Cellular Endocrinology, Vol. 124, No. 1-2, 29.11.1996, p. 43-50.

Research output: Contribution to journalArticle

Wenkert, D, Schoneberg, T, Merendino, JJ, Rodriguez Pena, MS, Vinitsky, R, Goldsmith, PK, Wess, J & Spiegel, AM 1996, 'Functional characterization of five V2 vasopressin receptor gene mutations', Molecular and Cellular Endocrinology, vol. 124, no. 1-2, pp. 43-50. https://doi.org/10.1016/S0303-7207(96)03926-3
Wenkert D, Schoneberg T, Merendino JJ, Rodriguez Pena MS, Vinitsky R, Goldsmith PK et al. Functional characterization of five V2 vasopressin receptor gene mutations. Molecular and Cellular Endocrinology. 1996 Nov 29;124(1-2):43-50. https://doi.org/10.1016/S0303-7207(96)03926-3
Wenkert, David ; Schoneberg, Torsten ; Merendino, John J. ; Rodriguez Pena, Maria Sol ; Vinitsky, Ruth ; Goldsmith, Paul K. ; Wess, Jurgen ; Spiegel, Allen M. / Functional characterization of five V2 vasopressin receptor gene mutations. In: Molecular and Cellular Endocrinology. 1996 ; Vol. 124, No. 1-2. pp. 43-50.
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