Mechanisms of Cell-surface Rerouting of an Endoplasmic Reticulum-retained Mutant of the Vasopressin V1b/V3 Receptor by a Pharmacological Chaperone

Abstract

Cell-surface expression and biological functions of several intracellular-retained G protein-coupled receptors are restored by membrane-permeable ligands called pharmacological chaperones. We have previously demonstrated that a mutation of the hydrophobic motif 341FNX2LLX3L350 in the C terminus of the human pituitary vasopressin V3 receptor (MUT V3R) led to it being retained in the endoplasmic reticulum (ER). Here, we establish the precise role of this motif and investigate whether SSR149415, a non-peptide V3R antagonist, behaves as a pharmacological chaperone for the ER-retained MUT V3R. The absence of the mutated receptor in the plasma membrane is linked to its prolonged association with the molecular chaperone calnexin in the ER and to its intensive degradation by the ubiquitin-proteasomal machinery. However, this is not because of a lack of oligomerization, as demonstrated by the presence of MUT V3R homodimers in the ER. Treatment with SSR149415 restores expression of the mutated receptor on the cell surface and its correct maturation, resulting into the functional recovery of its signaling properties. SSR149415 acts by stabilizing a native-like conformation of the V3R, reducing its association with calnexin and, thus, favoring a secretory pathway rather than the proteasomal degradation pathway. In conclusion, the FN(X)2LL(X)3L sequence is an important motif for the V3R conformation, and the misfolding resulting from its mutation alters the receptor export but can be reverted by SSR149415. Cell-surface expression and biological functions of several intracellular-retained G protein-coupled receptors are restored by membrane-permeable ligands called pharmacological chaperones. We have previously demonstrated that a mutation of the hydrophobic motif 341FNX2LLX3L350 in the C terminus of the human pituitary vasopressin V3 receptor (MUT V3R) led to it being retained in the endoplasmic reticulum (ER). Here, we establish the precise role of this motif and investigate whether SSR149415, a non-peptide V3R antagonist, behaves as a pharmacological chaperone for the ER-retained MUT V3R. The absence of the mutated receptor in the plasma membrane is linked to its prolonged association with the molecular chaperone calnexin in the ER and to its intensive degradation by the ubiquitin-proteasomal machinery. However, this is not because of a lack of oligomerization, as demonstrated by the presence of MUT V3R homodimers in the ER. Treatment with SSR149415 restores expression of the mutated receptor on the cell surface and its correct maturation, resulting into the functional recovery of its signaling properties. SSR149415 acts by stabilizing a native-like conformation of the V3R, reducing its association with calnexin and, thus, favoring a secretory pathway rather than the proteasomal degradation pathway. In conclusion, the FN(X)2LL(X)3L sequence is an important motif for the V3R conformation, and the misfolding resulting from its mutation alters the receptor export but can be reverted by SSR149415. The physiological actions of vasopressin (AVP) 2The abbreviations used are: AVP(Arg8)-vasopressinGPCRG protein-coupled receptorNDInephrogenic diabetes insipidusERendoplasmic reticulumQCquality controlWTwild-typeMUTmutantEGFPenhanced green fluorescent proteinRLucrenilla luciferaseEYFPenhanced yellow fluorescent proteinFACSfluorescence-activated cell sortingIPinositol phosphatesBRETbioluminescence resonance energy transferLSDleast significant difference. are mediated by three distinct seven-transmembrane domain G protein-coupled receptor (GPCR) subtypes, V1aR, V2R, and V1b/V3R (1Jard S. Adv. Exp. Med. Biol. 1998; 449: 1-13Crossref PubMed Scopus (54) Google Scholar, 2Birnbaumer M. Trends Endocrinol. Metab. 2000; 11: 406-410Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar, 3Thibonnier M. Coles P. Thibonnier A. Shoham M. Prog. Brain Res. 2002; 139: 179-196Crossref PubMed Scopus (85) Google Scholar). The V2 receptor has been extensively studied and mediates the antidiuretic effects of AVP in the renal collecting duct (4Birnbaumer M. Seibold A. Gilbert S. Ishido M. Barberis C. Antaramian A. Brabet P. Rosenthal W. Nature. 1992; 357: 333-335Crossref PubMed Scopus (490) Google Scholar). Natural mutations within the V2R gene are responsible for nephrogenic diabetes insipidus (NDI), and most of the 150 loss-of-function mutations so far identified cause retention of the receptor in the endoplasmic reticulum (ER) due to folding defects (5Oksche A. Rosenthal W. J. Mol. Med. 1998; 76: 326-337Crossref PubMed Scopus (117) Google Scholar, 6Morello J.P. Bichet D.G. Annu. Rev. Physiol. 2001; 63: 607-630Crossref PubMed Scopus (265) Google Scholar). The pituitary V3 receptor (7de Keyzer Y. Auzan C. Lenne F. Beldjord C. Thibonnier M. Bertagna X. Clauser E. FEBS Lett. 1994; 356: 215-220Crossref PubMed Scopus (191) Google Scholar, 8Sugimoto T. Saito M. Mochizuki S. Watanabe Y. Hashimoto S. Kawashima H. J. Biol. Chem. 1994; 269: 27088-27092Abstract Full Text PDF PubMed Google Scholar) is primarily involved in the cosecretory effect of AVP on corticotropin-releasing hormone-induced adrenocorticotropic hormone secretion (9Aguilera G. Rabadan-Diehl C. Regul. Pept. 2000; 96: 23-29Crossref PubMed Scopus (317) Google Scholar) and in the regulation of the stress response, as recently demonstrated by the receptor knock-out (10Wersinger S.R. Ginns E.I. O'Carroll A.M. Lolait S.J. Young III, W.S. Mol. Psychiatry. 2002; 7: 975-984Crossref PubMed Scopus (317) Google Scholar, 11Egashira N. Tanoue A. Higashihara F. Fuchigami H. Sano K. Mishima K. Fukue Y. Nagai H. Takano Y. Tsujimoto G. Stemmelin J. Griebel G. Iwasaki K. Ikeda T. Nishimura R. Fujiwara M. Neuropsychopharmacology. 2005; 30: 1996-2005Crossref PubMed Scopus (47) Google Scholar). To date, no natural mutation of this receptor has been associated with disease, but its structure-function relationships have been investigated using site-directed mutagenesis (12Derick S. Pena A. Durroux T. Wagnon J. Serradeil-Le Gal C. Hibert M. Rognan D. Guillon G. Mol. Endocrinol. 2004; 18: 2777-2789Crossref PubMed Scopus (20) Google Scholar, 13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). The mutation of the hydrophobic motif 341FNX2LLX3L350 in the proximal C terminus of the human V3R prevents its export from the ER to the cell surface (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). However, the precise role of this motif in folding, oligomerization, or its direct interaction with the export machinery remains unknown. (Arg8)-vasopressin G protein-coupled receptor nephrogenic diabetes insipidus endoplasmic reticulum quality control wild-type mutant enhanced green fluorescent protein renilla luciferase enhanced yellow fluorescent protein fluorescence-activated cell sorting inositol phosphates bioluminescence resonance energy transfer least significant difference. The transport from the ER of newly synthesized proteins is strictly regulated by a conformation-based process called quality control (QC) (14Trombetta E.S. Parodi A.J. Annu. Rev. Cell Dev. Biol. 2003; 19: 649-676Crossref PubMed Scopus (364) Google Scholar, 15Kleizen B. Braakman I. Curr. Opin. Cell Biol. 2004; 16: 343-349Crossref PubMed Scopus (366) Google Scholar). During QC, different molecular chaperone proteins transiently interact with the nascent polypeptide chain, thus ensuring its proper conformation. Properly folded proteins are directed to the secretory pathway via the export machinery. Misfolded proteins remain persistently associated with the molecular chaperones, are retained in the ER, and are finally degraded by the proteasome after ubiquitination (16Tsai B. Ye Y. Rapoport T.A. Nat. Rev. Mol. Cell Biol. 2002; 3: 246-255Crossref PubMed Scopus (552) Google Scholar, 17Glickman M.H. Ciechanover A. Physiol. Rev. 2002; 82: 373-428Crossref PubMed Scopus (3377) Google Scholar, 18Bonifacino J.S. Weissman A.M. Annu. Rev. Cell Dev. Biol. 1998; PubMed Scopus Google Scholar). has been for the receptor M. A. S. P. M. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). an ER and in the and folding of secretory and several as V2R J.P. A. A. M. Bichet D.G. M. 2001; PubMed Scopus Google receptor S. A. J. J. 2002; 269: PubMed Scopus Google hormone and hormone receptor Y. 1998; 139: PubMed Scopus Google Scholar). calnexin for with of the V2R, to retention in the ER J.P. A. A. M. Bichet D.G. M. 2001; PubMed Scopus Google Scholar). misfolding defects is a in that the ER retention of mutated is the of membrane-permeable called pharmacological that to the protein and it to be M. J. 2000; PubMed Scopus Google Scholar, J.P. Bichet D.G. M. Trends 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, A. 2004; PubMed Scopus Google Scholar). a and a V2R antagonist, have been to the expression and biological functions of the mutant involved in and of several V2R involved in Y. S. K. S. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar, J.P. A. A. M. S. D. Bichet D.G. M. J. 2000; PubMed Scopus Google Scholar). However, it is not whether pharmacological the newly synthesized the or degradation of the is that oligomerization, has been recently identified for the vasopressin receptors S. Durroux T. B. A. M.A. M. R. Barberis C. M. Mol. Endocrinol. 2003; PubMed Scopus Google Scholar) and is to be important in export A. S. M. S. M. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). Here, we have the effects of SSR149415, the non-peptide vasopressin V3R Gal C. Wagnon J. J. Griebel G. C. Guillon G. Barberis C. G. P. D. M. R. B. J.P. G. J. Exp. 2002; PubMed Scopus Google on the previously ER-retained V3R mutant (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). We that SSR149415 behaves as a pharmacological chaperone and maturation, and the signaling functions of the is due to with the chaperone calnexin and proteasomal degradation from or for molecular from from and (AVP) from with and in cell from and from We used the and V3 wild-type and mutated receptor into the and the as previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). To the the sequence of the wild-type V3R by using the and and the and of the into the of The expression for V3R and MUT V3R with terminus and the C terminus called and the the previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google from the sequence using and a this a the the the and of the V3R used to the in the The by the of wild-type V3R by the mutated in the and V3R and after of the with or and of the V3R into the of or wild-type the mutant and by of a the and the V3R sequence by a of the The sequence from using the site-directed mutagenesis The a from and has been previously A. S. M. S. M. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). Cell and in with and in with III, and transiently using the and the and transiently using the to the The with the non-peptide V3R SSR149415 from Serradeil-Le Gal of by for or as in the after Cell transiently with the or the with and and by using a as previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). of V3 on transiently using as a as previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). We inositol phosphates in transiently as previously Y. M. J. C. R. Google Scholar). with AVP and of receptors and using the and on and transiently and as previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). using a and and of the V3R and MUT V3R on transiently as previously (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). of calnexin and to using a or a and in or the for in and and with or of in cell in the for or the in and for of in with and and finally in The of V3 receptors as V3 receptors with calnexin by a and protein and by a and protein The receptors on and to the of association of V3R with of the V3R and and the V3R and in in the absence and or presence and of SSR149415. by with for by the of using and protein by and protein The are by an of the association the of the V3R and the MUT V3R and calnexin with SSR149415 and or and by of the from with being the the for The to a are the of three different with and after in The and in and in a a of and using a that the and to be and using The as the the and of the and proteins and the and of the protein The in to the by We and for to the of expression of the and and to that the of with different of for a of or are as the We used of to significant by of a using the least significant using the with the of the V3R and MUT a functional of the V3 receptor and a mutated V3R in a proximal hydrophobic motif by in mutant (MUT V3R) previously not to be to the cell surface but to be retained in the ER (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). We the terminus of the human V3 receptor to to and the receptor The of an to the C terminus of the to receptor expression and by The of the the of V3R expression with the (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). the of the and V3R for and of expression the plasma membrane and and receptors We that the mutant receptor to AVP the cell surface of the presence of the and that this not and We used to the lack of expression of the mutant MUT V3R and its in the endoplasmic reticulum in thus (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). SSR149415 Treatment Cell-surface of the MUT transiently the V3R or V3R or not for with SSR149415, a V3R We in by using the of to We that with SSR149415 for restored the of expression of the MUT V3R to that for the V3R The of this effect for SSR149415 SSR149415 the export of V3R with the We in using In the absence of SSR149415, the MUT V3R retained in the ER and from the and the plasma membrane In the presence of SSR149415, the MUT V3R retained in the ER, the secretory as by its presence in the and finally the cell surface SSR149415 Treatment the of the MUT used in the V3R or MUT V3R to the role of SSR149415 on receptor and The V3R synthesized as a receptor that to a receptor have that the is to and and, to an the is to and to and, to a receptor (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). Treatment with SSR149415 enhanced the and the of the V3R as by the of the and the of the for of proteins In the absence of SSR149415, the MUT V3R a to the protein and of with the V3R of In the presence of SSR149415, the MUT V3R to a with the from the a to the V3R and that SSR149415 the MUT V3R from a to a secretory pathway. SSR149415 Treatment the of the MUT investigated whether SSR149415 restores the signaling of the MUT V3R in to the of its to the plasma We transiently the V3R or V3R with SSR149415 for The to the and with AVP to the of via the V3 receptor with the in the presence or the absence of SSR149415 However, the of AVP in the absence of SSR149415 than in its presence AVP and the in not AVP to via the MUT V3R after with SSR149415 In the presence of SSR149415, the of AVP for the the for the MUT V3R and V3R that SSR149415 restores the signaling of the mutant The of SSR149415 for this with that for expression of the mutant that SSR149415 behaves as a pharmacological expression and the and signaling of the human MUT V3R. a role for the FN(X)2LL(X)3L We whether SSR149415 the oligomerization, prevents the or the mutant V3 of the V3R by SSR149415 that in the ER and is a process for receptors to the cell surface S. M. 2004; PubMed Scopus Google Scholar). and has been demonstrated for the and the V2 receptors S. Durroux T. B. A. M.A. M. R. Barberis C. M. Mol. Endocrinol. 2003; PubMed Scopus Google Scholar) but not for the vasopressin V3 we studied the of V3R in using the We a for the V3R, we not V3R and an that V3R as We the for the MUT V3R, that of this receptor in the ER and that the mutation not cause an We that SSR149415 the of the V3R in a SSR149415 not the for the MUT V3R the that its role the mutant receptor to the plasma membrane is not linked to We to investigate whether SSR149415 the of to and, the and or whether it the conformation of The to the and the a of the of the for the V3R, SSR149415 the the SSR149415 and with We the for the MUT V3R SSR149415 and with that SSR149415 the conformation rather than the process SSR149415 Treatment of the from the ER proteins are degraded by the ubiquitin-proteasomal J.S. Weissman A.M. Annu. Rev. Cell Dev. Biol. 1998; PubMed Scopus Google Scholar). we studied whether this is involved in V3R and is regulated by SSR149415. using we that SSR149415 the expression of V3R and MUT V3R and in a for the V3R) it is that SSR149415 acts as a we that the degradation pathway and in the ER and the plasma to an in of the We whether the V3R and MUT V3R are to the ubiquitin-proteasomal by the of into V3R. the V3R as demonstrated by the presence of a We that the MUT V3R that it However, after a with SSR149415, the ubiquitination of the V3R The the ubiquitination of the MUT V3R. not due to in the of V3R, as in that SSR149415 prevents proteasomal degradation of the V3 SSR149415 Treatment of the V3R with the retention of mutated proteins has been linked to prolonged interaction of the mutant receptor with the molecular of the ER. we V3R and V3R and the physiological chaperone calnexin with to whether calnexin can interact with the vasopressin V3 We the of the calnexin to the of receptor We that the of receptors associated with calnexin for the MUT V3R than for the V3R, that calnexin a role in the retention of the mutated receptor in the ER SSR149415 the interaction of calnexin with the V3R and the MUT V3R by and by stabilizing the conformation of the V3 receptor We the of the association of calnexin and the newly synthesized V3 receptor and investigated the role of SSR149415 in this process by by the of the receptor with calnexin The of the MUT V3R and calnexin than that of the V3R of and that calnexin a role in the mutated receptor in the ER. In the presence of SSR149415, the of of calnexin and the receptor for the V3R of and the MUT V3R of that SSR149415 a native-like conformation that a newly synthesized receptor to to a prolonged interaction with We have previously that mutations in the hydrophobic sequence FN(X)2LL(X)3L in the proximal C terminus of the V3 receptor in this receptor being retained in the ER (13Robert J. Clauser E. Petit P.X. Ventura M.A. J. Biol. Chem. 2005; 280: 2300-2308Abstract Full Text Full Text PDF PubMed Scopus (67) Google Scholar). However, it not whether this motif is a for ER export or is for folding of the In the we the of the ER-retained V3R mutant with the quality control machinery using a non-peptide of the V3R. behaves as a pharmacological the cell and have been to several receptors in the ER and to maturation, and biological from the that the mutant V3 receptor has a folding it in a for with and is with the wild-type During in the ER, proteins interact with several molecular that the correct folding of the being to the export machinery. the ER with to the nascent of several of the as the V2 J.P. A. A. M. Bichet D.G. M. 2001; PubMed Scopus Google and hormone Y. 1998; 139: PubMed Scopus Google Scholar) interact with this several and the conformation of the and the the to the association of this a the MUT V3R, the mutations are in the C terminus of the V3R, the calnexin is in the ER a rather than a folding that the FN(X)2LL(X)3L motif is involved in the folding of the receptor rather than in its are with that the motif of the V2R is important for the correct folding of the with the involved in a hydrophobic interaction with from the G. R. A. C. Rosenthal W. R. Mol. 2000; Google Scholar). V2 receptor and have been to be retained in the ER due to prolonged association with calnexin J.P. A. A. M. Bichet D.G. M. 2001; PubMed Scopus Google Scholar). the retention of the in the ER by molecular is responsible for its to the degradation pathway. degradation is by the of the protein in the its and its degradation by the proteasome (16Tsai B. Ye Y. Rapoport T.A. Nat. Rev. Mol. Cell Biol. 2002; 3: 246-255Crossref PubMed Scopus (552) Google Scholar, 17Glickman M.H. Ciechanover A. Physiol. Rev. 2002; 82: 373-428Crossref PubMed Scopus (3377) Google Scholar, 18Bonifacino J.S. Weissman A.M. Annu. Rev. Cell Dev. Biol. 1998; PubMed Scopus Google Scholar). The mutant V3 receptor is it are for the receptor M. A. S. P. M. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). We demonstrated that SSR149415, a non-peptide and of the V3 is to the maturation, and signaling of the ER-retained SSR149415 to the of pharmacological chaperones. are to to a a and to it in a conformation with a energy and in a pharmacological have been to the conformation of vasopressin V2 J.P. A. A. M. S. D. Bichet D.G. M. J. 2000; PubMed Scopus Google Scholar, S. B. A. J. G. R. M. R. Rosenthal W. A. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google hormone M. E. J. D.G. J. Exp. 2003; PubMed Scopus Google Y. S. K. S. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google Mol. 2003; PubMed Scopus Google and human hormone receptor J. S.J. Y. Mol. Endocrinol. 2005; 19: PubMed Scopus Google Scholar) mutant receptors and to expression on the cell The by the pharmacological chaperone restores the expression of the V3 receptor on the cell surface is linked to the of protein conformation. molecular of the interaction the human V3R and SSR149415 has recently been on the of (12Derick S. Pena A. Durroux T. Wagnon J. Serradeil-Le Gal C. Hibert M. Rognan D. Guillon G. Mol. Endocrinol. 2004; 18: 2777-2789Crossref PubMed Scopus (20) Google Scholar). this it has been that the are for the of the interaction with and hydrophobic with the SSR149415 the correct of the receptor that are by the SSR149415 the of the receptor with calnexin and its degradation by the The of is and the protein a can the receptor from an pathway to a secretory restores the receptor we that the mutated sequence of the V3 receptor the conformation of the but is that the pharmacological chaperone the conformation of the receptor and a motif that is in the wild-type receptor but is in the mutant motif interact with a an protein or a protein of the export machinery. and have been to the correct folding of receptors H. M. H. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). and have been to as molecular for M.A. Full Text PDF PubMed Scopus Google Scholar, T.A. Nature. PubMed Scopus Google Scholar). is that the pharmacological chaperone of the quality control or the for the receptor to be associated to an the pharmacological chaperone to the expression of the receptor on the cell for the that the not the of the We using the the wild-type V3 the mutant receptor homodimers and that SSR149415 not the that and expression of the mutant on the cell the V3 receptor can homodimers its in the ER. has recently been demonstrated for as the S. A. E. S. D. M. M. S. A. 2000; Google the hormone A. G. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google the T. A. FEBS Lett. 2001; PubMed Scopus Google or the and the vasopressin and V2 S. Durroux T. B. A. M.A. M. R. Barberis C. M. Mol. Endocrinol. 2003; PubMed Scopus Google Scholar) it has been demonstrated that of the receptor in the ER is a for A. S. M. S. M. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). an be by the pharmacological the retention of the MUT V3R in the ER, that this is not the We that SSR149415 to the of the wild-type V3 receptor in a of the that the with no in the the the that SSR149415 a of the receptor by the the a role of the pharmacological we that SSR149415 to the of the wild-type V3 receptor and its expression on the cell that this a pharmacological chaperone than for the mutant that the V3R is but retained in the ER. export is by pharmacological chaperones, has been for the wild-type V2 receptor S. B. A. J. G. R. M. R. Rosenthal W. A. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google the wild-type receptor M. A. P. M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, M. S. A. J.P. M. J. 2002; PubMed Scopus Google and the wild-type hormone receptor A. 2004; PubMed Scopus Google Scholar). is in with in the pharmacological or not the of the wild-type human V2 receptor J.P. A. A. M. S. D. Bichet D.G. M. J. 2000; PubMed Scopus Google the wild-type receptor Mol. 2003; PubMed Scopus Google or the wild-type Y. S. K. S. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google wild-type have a in in the ER, is a in is that in proteins as the previously proteins this In conclusion, the non-peptide SSR149415 is a pharmacological chaperone for the vasopressin V3 receptor and is to the maturation, and signaling of a ER-retained mutant V3 the of pharmacological in of due to ER retention of the mutated the pharmacological chaperone to the mutated protein but its functional or We Serradeil-Le Gal for the of SSR149415, Petit for the and for the from the of the for the and for the and