Variant amino acids in the extracellular loops of murine and human vasopressin V2 receptors account for differences in cell surface expression and ligand affinity

Molecular Endocrinology
Alexander OkscheWalter Rosenthal

Abstract

Cloning and sequencing of the murine chromosomal region XB harboring the murine vasopressin V(2) receptor (mV(2)R) gene and comparison with the orthologous human Xq28 region harboring the human vasopressin V(2) receptor (hV(2)R) revealed conservation of the genomic organization and a high degree of sequence identity in the V(2)R coding regions. Despite an identity of 87% of the amino acid sequences, both receptors show marked functional differences upon stable expression in Chinese hamster ovary cells: the mV(2)R displayed a 5-fold higher affinity for [(3)H]AVP than the human ortholog; similar differences were found for the AVP-mediated activation of adenylyl cyclase. Saturation binding experiments with transiently transfected intact COS.M6 cells showed that the mV(2)R was 3- to 5-fold less abundantly expressed at the cell surface than the hV(2)R. Laser scanning microscopy of fusion proteins consisting of the V(2)Rs and green fluorescent protein (GFP) (mV(2)R/GFP, hV(2)R/GFP) demonstrated that the hV(2)R/GFP was efficiently transported to the plasma membrane, whereas the mV(2)R/GFP was localized mainly within the endoplasmic reticulum. Chimeric hV(2)Rs, in which the first and/or second extracellular loop(s) were replaced by the...Continue Reading

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Citations

Sep 5, 2009·Journal of the American Society of Nephrology : JASN·Frédéric Jean-AlphonseChristiane Mendre
Dec 15, 2004·Journal of Pharmacological and Toxicological Methods·K H S ArunP Ramarao
May 23, 2006·Seminars in Nephrology·Friedericke Quittnat, Peter Gross
Oct 9, 2009·American Journal of Physiology. Renal Physiology·Mia FaerchThomas J Corydon
Jun 8, 2007·American Journal of Physiology. Renal Physiology·Robert A FentonJeppe Praetorius
Mar 26, 2014·Endocrine Reviews·Ya-Xiong Tao, P Michael Conn
Sep 13, 2002·The Journal of Biological Chemistry·Evelina GrantcharovaAlexander Oksche
May 12, 2020·Molecular and Cellular Endocrinology·Ya-Xiong Tao

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