The D136A mutation of the V<sub>2</sub> vasopressin receptor induces a constitutive activity which permits discrimination between antagonists with partial agonist and inverse agonist activities

Morin, Denis; Cotte, Nathalie; Balestre, Marie-Noëlle; Mouillac, Bernard; Manning, Maurice; Breton, Christophe; Barberis, Claude

doi:10.1016/s0014-5793(98)01585-3

Cited by 73 publications

(60 citation statements)

References 35 publications

(42 reference statements)

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“…These results demonstrate that NAPol-reconstituted V2R interacts with and efficiently activates Gs protein in a ligand-dependent manner. Importantly, the efficacy profiles of the ligands correlated well with those observed in living cells (18,21).…”

Section: Resultssupporting

confidence: 55%

“…To determine the functionality of NAPol-reconstituted V2R, we first measured ligand-induced incorporation of [ 35 S]GTPγS to purified Gαs proteins (1:2.5 receptor/G protein molar ratio). This assay was done using two full agonists for the Gs pathway, the endogenous hormone AVP and the biased ligand MCF14 (18), as well as with a Gs inverse agonist (SR121463) (21) (Fig. 2A).…”

Section: Resultsmentioning

confidence: 99%

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Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Rahmeh

Damian

Cottet

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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173

137

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G protein-coupled receptors (GPCRs) are seven-transmembrane proteins that mediate most cellular responses to hormones and neurotransmitters, representing the largest group of therapeutic targets. Recent studies show that some GPCRs signal through both G protein and arrestin pathways in a ligand-specific manner. Ligands that direct signaling through a specific pathway are known as biased ligands. The arginine-vasopressin type 2 receptor (V2R), a prototypical peptide-activated GPCR, is an ideal model system to investigate the structural basis of biased signaling. Although the native hormone arginine-vasopressin leads to activation of both the stimulatory G protein (Gs) for the adenylyl cyclase and arrestin pathways, synthetic ligands exhibit highly biased signaling through either Gs alone or arrestin alone. We used purified V2R stabilized in neutral amphipols and developed fluorescence-based assays to investigate the structural basis of biased signaling for the V2R. Our studies demonstrate that the Gs-biased agonist stabilizes a conformation that is distinct from that stabilized by the arrestin-biased agonists. This study provides unique insights into the structural mechanisms of GPCR activation by biased ligands that may be relevant to the design of pathway-biased drugs.

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Section: Resultssupporting

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Rahmeh

Damian

Cottet

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

173

137

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“…The DRY motif has played a pivotal role in the signal transduction pathway of GPCRs (30 -32). Mutations of the aspartate residue are reported to lead to constitutive activity for some GPCRs (31)(32)(33). Similar mutation in the homologous position in the rat B 1 receptor failed to induce detectable constitutive activity.…”

Section: Discussionmentioning

confidence: 89%

Overexpression of Kinin B1 Receptors Induces Hypertensive Response to Des-Arg9-bradykinin and Susceptibility to Inflammation

Yin

Agata

et al. 2003

Journal of Biological Chemistry

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We demonstrated that rat kinin B 1 receptors displayed a ligand-independent constitutive activity, assessed through inositol phosphate production in transiently or stably transfected human embryonic kidney 293A cells. Substitution of Ala for Asn 130 in the third transmembrane domain resulted in additional constitutive activation of the B 1 receptor. The constitutively active mutant N130A receptor could be further activated by the B 1 receptor agonist des-Arg 9 -bradykinin. To gain insights into the physiological function of the B 1 receptor, we have generated transgenic mice overexpressing wild-type and constitutively active mutant receptors under the control of human cytomegalovirus immediately early gene enhancer/promoter. The rat B 1 receptor transgene expression was detected in the aorta, brain, heart, lung, liver, kidney, uterus, and prostate of transgenic mice by reverse transcriptionpolymerase chain reaction/Southern blot analysis. Transgenic mice were fertile and normotensive. Overexpression of B 1 receptors exacerbated paw edema induced by carrageenan and rendered transgenic mice more susceptible to lipopolysaccharide-induced endotoxic shock. Interestingly, the hemodynamic response to kinins was altered in transgenic mice, with des-Arg 9 -bradykinin inducing blood pressure increase when intravenously administered. Our study supports an important role for B 1 receptors in modulating inflammatory responses and for the first time demonstrates that B 1 receptors mediate a hypertensive response to des-Arg 9 -bradykinin.Kinin peptides are released from kininogen precursors by the action of kallikreins in response to tissue injury (1). Kinins induce smooth muscle contraction, vasodilation, increased vascular permeability, and pain (1). Kinins exert their effects through selective activation of two seven-transmembrane domain (TMD) 1 G protein-coupled receptors (GPCRs), B 1 and B 2 (2-4). The B 2 receptor is constitutively expressed, mediating the actions of intact kinins, bradykinin (BK) in rodents and Lys-BK or kallidin in humans (2). In contrast, the B 1 receptor is expressed at very low levels in normal tissues in most animal species but is induced under the influence of inflammation or exposure of tissues to noxious stimuli, mediating the effects of the carboxypeptidase metabolites of intact kinins, des-Arg 9 -BK (DABK), and des-Arg 10 -kallidin (2). The cellular responses of kinin receptors to agonists are transduced primarily via coupling to either G q protein, which in turn activates phospholipase C to stimulate inositol phosphate production, or the G i protein, acting through phospholipase A 2 to stimulate arachidonic acid pathway (5, 6).Over the past years, transgenic and gene-targeting technologies associated with molecular biology tools have provided important knowledge concerning the role of kinin receptors in vivo. Transgenic mice expressing the human B 2 receptor under the control of the Rous sarcoma virus 3Ј-long terminal repeat promoter were hypotensive compared with control littermates (7). Adm...

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“…at the exoloop 2 or intraloop 2, which share 76 and 62% amino acid identity with the corresponding domains in the rGnRHR); in this setting, the particular structure of the rGnRHR antagonist might ultimately determine a preference for binding to the active receptor conformation of the hGnRHR and to both the inactive and active conformations of the rat counterpart, as would be expected for antagonist analogues. In fact, in a number of G protein-coupled receptors, including the GnRHR, mutations or species-dependent structural differences in either transmembrane domains (Strader et al 1989, Claude et al 1996, Noda et al 1996, Groblewski et al 1997, Han et al 1997, Spivak et al 1997, intraloops (Morin et al 1998, Wurch et al 1999 or exoloops (Sun et al 2001) may be limiting, to a variable extent, the level of maximal stimulation. It should be emphasized, however, that the effects of the compounds tested may be cell-specific and that depending on the relative levels of receptor, G proteins and other downstream effectors present in a particular tissue in vivo (i.e.…”

Section: Discussionmentioning

confidence: 99%

Identification of new gonadotrophin-releasing hormone partial agonists

Leaños‐Miranda

Ulloa‐Aguirre²,

Cervini³

et al. 2006

Journal of Endocrinology

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GnRH agonists or antagonists are currently utilized as therapeutic agents in a number of diseases. A side-effect of prolonged treatment with GnRH analogues is hypoestrogenism. In this study, we tested the in vitro potency of different GnRH analogues originally found to be partial agonists (i.e. analogues with decreased efficacy for activating or stimulating their cognate receptor) as well as novel analogues, to identify compounds that might potentially be useful for partial blockade of gonadotrophin release.

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The D136A mutation of the V₂ vasopressin receptor induces a constitutive activity which permits discrimination between antagonists with partial agonist and inverse agonist activities

Abstract: The substitution, in the human V 2 vasopressin receptor, of the aspartate at position 136 by alanine leads to agonist-independent activation of this mutant V 2 receptor.

Cited by 73 publications

References 35 publications

Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Overexpression of Kinin B1 Receptors Induces Hypertensive Response to Des-Arg9-bradykinin and Susceptibility to Inflammation

Identification of new gonadotrophin-releasing hormone partial agonists

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The D136A mutation of the V2 vasopressin receptor induces a constitutive activity which permits discrimination between antagonists with partial agonist and inverse agonist activities

Abstract: The substitution, in the human V 2 vasopressin receptor, of the aspartate at position 136 by alanine leads to agonist-independent activation of this mutant V 2 receptor.

Cited by 73 publications

References 35 publications

Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Structural insights into biased G protein-coupled receptor signaling revealed by fluorescence spectroscopy

Overexpression of Kinin B1 Receptors Induces Hypertensive Response to Des-Arg9-bradykinin and Susceptibility to Inflammation

Identification of new gonadotrophin-releasing hormone partial agonists

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The D136A mutation of the V₂ vasopressin receptor induces a constitutive activity which permits discrimination between antagonists with partial agonist and inverse agonist activities