This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sadeghi, H. M. Articles by Birnbaumer, M. Search for Related Content PubMed PubMed Citation Articles by Sadeghi, H. M. Articles by Birnbaumer, M.

An X-Linked NDI Mutation Reveals a Requirement for Cell Surface V2R Expression

Department of Anesthesiology and Molecular Biology Institute, University of California Los Angeles School of Medicine, Los Angeles, California 90095

Function and biochemical properties of the V2 vasopressin receptor (V2R) mutant R337ter, identified in patients suffering from X-linked recessive nephrogenic diabetes insipidus, were investigated by expression in COS.M6 or HEK293 cells. Binding assays and measurements of adenylyl cyclase activity failed to detect function for the truncated receptor, although metabolic labeling demonstrated normal levels of protein synthesis. ELISA assays performed on cells expressing the receptors tagged at the amino terminus with the HA epitope failed to detect V2R R337ter on the plasma membrane. Treatment with endoglycosidase H revealed that the receptor was present only as a precursor form because the mature R337ter V2R, resistant to endoglycosidase H treatment, was not detected. The precursor of V2R-R337ter had a longer half-life than that of the wild type V2R, suggesting that arrested maturation may slow the degradation of the precursor. Unrelated experiments had demonstrated that V2R-G345ter, containing eight additional amino acids, was expressed on the plasma membrane and functioned normally. Receptor truncations longer than 337ter revealed that four of the eight amino acids identified initially provided the minimum length required for the protein to acquire cell surface expression. This was shown by the production of mature receptor (V2R-341ter) detectable in SDS-PAGE, which mediated arginine vasopressin stimulation of adenylyl cyclase activity and bound ligand. In addition, the identity of amino acid 340 was found to play a role in this phenomenon. In conclusion, these data demonstrate that the V2R R337ter is nonfunctional because it does not reach the plasma membrane and that the minimal protein length required for translocation of the V2R to the cell surface is sufficient to confer function to the receptor protein. They also suggest the existence of a protein quality control in the endoplasmic reticulum independent of glycosylation.

This article has been cited by other articles:

				P. M. Conn, A. Ulloa-Aguirre, J. Ito, and J. A. Janovick G Protein-Coupled Receptor Trafficking in Health and Disease: Lessons Learned to Prepare for Therapeutic Mutant Rescue in Vivo Pharmacol. Rev., September 1, 2007; 59(3): 225 - 250. [Abstract] [Full Text] [PDF]

				S. K. Shenoy Seven-Transmembrane Receptors and Ubiquitination Circ. Res., April 27, 2007; 100(8): 1142 - 1154. [Abstract] [Full Text] [PDF]

				J. H. Robben, N. V. A. M. Knoers, and P. M. T. Deen Cell biological aspects of the vasopressin type-2 receptor and aquaporin 2 water channel in nephrogenic diabetes insipidus. Am J Physiol Renal Physiol, August 1, 2006; 291(2): F257 - F270. [Abstract] [Full Text] [PDF]

				J. Robert, E. Clauser, P. X. Petit, and M. A. Ventura A Novel C-terminal Motif Is Necessary for the Export of the Vasopressin V1b/V3 Receptor to the Plasma Membrane J. Biol. Chem., January 21, 2005; 280(3): 2300 - 2308. [Abstract] [Full Text] [PDF]

				A. Schulz, K. Sangkuhl, T. Lennert, M. Wigger, D. A. Price, A. Nuuja, A. Gruters, G. Schultz, and T. Schoneberg Aminoglycoside Pretreatment Partially Restores the Function of Truncated V2 Vasopressin Receptors Found in Patients with Nephrogenic Diabetes Insipidus J. Clin. Endocrinol. Metab., November 1, 2002; 87(11): 5247 - 5257. [Abstract] [Full Text] [PDF]

				M. Birnbaumer The V2 vasopressin receptor mutations and fluid homeostasis Cardiovasc Res, August 15, 2001; 51(3): 409 - 415. [Abstract] [Full Text] [PDF]

				J. K. VanSlyke, S. M. Deschenes, and L. S. Musil Intracellular Transport, Assembly, and Degradation of Wild-Type and Disease-linked Mutant Gap Junction Proteins Mol. Biol. Cell, June 1, 2000; 11(6): 1933 - 1946. [Abstract] [Full Text]

				G. Krause, R. Hermosilla, A. Oksche, C. Rutz, W. Rosenthal, and R. Schülein Molecular and Conformational Features of a Transport-Relevant Domain in the C-Terminal Tail of the Vasopressin V2 Receptor Mol. Pharmacol., February 1, 2000; 57(2): 232 - 242. [Abstract] [Full Text]

				A. Schulz, R. Grosse, G. Schultz, T. Gudermann, and T. Schoneberg Structural Implication for Receptor Oligomerization from Functional Reconstitution Studies of Mutant V2 Vasopressin Receptors J. Biol. Chem., January 28, 2000; 275(4): 2381 - 2389. [Abstract] [Full Text] [PDF]

				R. Schülein, R. Hermosilla, A. Oksche, M. Dehe, B. Wiesner, G. Krause, and W. Rosenthal A Dileucine Sequence and an Upstream Glutamate Residue in the Intracellular Carboxyl Terminus of the Vasopressin V2 Receptor Are Essential for Cell Surface Transport in COS.M6 Cells Mol. Pharmacol., September 1, 1998; 54(3): 525 - 535. [Abstract] [Full Text]

				A. C. Latronico, Y. Chai, I. J.P. Arnhold, X. Liu, B. B. Mendonca, and D. L. Segaloff A Homozygous Microdeletion in Helix 7 of the Luteinizing Hormone Receptor Associated with Familial Testicular and Ovarian Resistance Is Due to Both Decreased Cell Surface Expression and Impaired Effector Activation by the Cell Surface Receptor Mol. Endocrinol., March 1, 1998; 12(3): 442 - 450. [Abstract] [Full Text]

				R. Schulein, K. Zuhlke, G. Krause, and W. Rosenthal Functional Rescue of the Nephrogenic Diabetes Insipidus-causing Vasopressin V2 Receptor Mutants G185C and R202C by a Second Site Suppressor Mutation J. Biol. Chem., March 9, 2001; 276(11): 8384 - 8392. [Abstract] [Full Text] [PDF]