Evolutionary action and structural basis of the allosteric switch controlling β2AR functional selectivity

Schönegge, Anne–Marie; Gallion, Jonathan; Picard, Louis-Philippe; Wilkins, Angela D.; Gouill, Christian Le; Audet, Martin; Stallaert, Wayne; Lohse, Martin J.; Kimmel, Marek; Lichtarge, Olivier; Bouvier, Michel

doi:10.1038/s41467-017-02257-x

Cited by 67 publications

(74 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The NPxxY motif plays an essential role on the receptor downregulation via clathrin-mediated endocytosis in several GPCRs including V2R 43 . In addition, in the family of beta 2-adrenergic receptors, it was found fundamental also for G-protein signalling 44,45 . The identified W323_I324insR mutation of V2R alters the conserved NPxxY motif by inserting an arginine at position 324 and replacing tyrosine with isoleucine at position 325.…”

Section: Discussionmentioning

confidence: 99%

Characterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation

Prosperi

Suzumoto

Marzuillo

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Nephrogenic diabetes insipidus (NDI) is a rare tubulopathy characterized by urinary concentration defect due to renal resistance to vasopressin. Loss-of-function mutations of vasopressin V2 receptor (V2R) gene (AVPR2) is the most common cause of the disease. We have identified five novel mutations L86P, R113Q, C192S, M272R, and W323_I324insR from NDI-affected patients. Functional characterization of these mutants revealed that R113Q and C192S were normally localized at the basolateral membrane of polarized Madin-Darby Canine Kidney (MDCK) cells and presented proper glycosylation maturation. On the other side, L86P, M272R, and W323_I324insR mutants were retained in endoplasmic reticulum and exhibited immature glycosylation and considerably reduced stability. All five mutants were resistant to administration of vasopressin analogues as evaluated by defective response in cAMP release. In order to rescue the function of the mutated V2R, we tested VX-809, sildenafil citrate, ibuprofen and tolvaptan in MDCK cells. Among these, tolvaptan was effective in rescuing the function of M272R mutation, by both allowing proper glycosylation maturation, membrane sorting and response to dDAVP. These results show an important proof of concept for the use of tolvaptan in patients affected by M272R mutation of V2R causing NDI.

show abstract

Section: Discussionmentioning

confidence: 99%

Characterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation

Prosperi

Suzumoto

Marzuillo

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Endocytosis trafficking plays a key role in controlling the activity of β2AR [36]. A large body of evidence from many investigators supports the notion that β2AR plays a carcinogenic role dependent on receptor endocytosis [37].…”

Section: Discussionmentioning

confidence: 99%

Mammalian Eps15 homology domain 1 potentiates angiogenesis of non-small cell lung cancer by regulating β2AR signaling

Wang

Xing

Meng

et al. 2019

J Exp Clin Cancer Res

View full text Add to dashboard Cite

Background: Non-small cell lung cancer (NSCLC) is a devastating disease with a heterogeneous prognosis, and the molecular mechanisms underlying tumor progression remain elusive. Mammalian Eps15 homology domain 1 (EHD1) plays a promotive role in tumor progression, but its role in cancer angiogenesis remains unknown. This study thus explored the role of EHD1 in angiogenesis in NSCLC. Methods: The changes in angiogenesis were evaluated through human umbilical vein endothelial cell (HUVEC) proliferation, migration and tube formation assays. The impact of EHD1 on β2-adrenoceptor (β2AR) signaling was evaluated by Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and enzymelinked immunosorbent assay (ELISA). The interaction between EHD1 and β2AR was confirmed by immunofluorescence (IF) and coimmunoprecipitation (Co-IP) experiments, and confocal microscopy immunofluorescence studies revealed that β2AR colocalized with the recycling endosome marker Rab11, which indicated β2AR endocytosis. Xenograft tumor models were used to investigate the role of EHD1 in NSCLC tumor growth. Results: The microarray analysis revealed that EHD1 was significantly correlated with tumor angiogenesis, and lossand gain-of-function experiments demonstrated that EHD1 potentiates HUVEC proliferation, migration and tube formation. EHD1 knockdown inhibited β2AR signaling activity, and EHD1 upregulation promoted vascular endothelial growth factor A (VEGFA) and β2AR expression. Interestingly, EHD1 interacted with β2AR and played a novel and critical role in β2AR endocytic recycling to prevent receptor degradation. Aberrant VEGFA or β2AR expression significantly affected EHD1-mediated tumor angiogenesis. The proangiogenic role of EHD1 was confirmed in xenograft tumor models, and immunohistochemistry (IHC) analysis confirmed that EHD1 expression was positively correlated with VEGFA expression, microvessel density (MVD) and β2AR expression in patient specimens. Conclusion: Collectively, the data obtained in this study suggest that EHD1 plays a critical role in NSCLC angiogenesis via β2AR signaling and highlight a potential target for antiangiogenic therapy.

show abstract

“…Observations of distinct ligands acting on the same receptor differentially activating different subsets of signaling effectors in cells have led to the idea that receptors can be conformationally stabilized by ligands in multiple discrete active signaling states (6), a notion that is supported by receptor structural studies (7,8). The overall signaling profile of a particular GPCR-ligand combination can also be modified by naturally occurring polymorphisms and engineered mutations, which also affect the conformational signaling landscape of the ligand-bound receptor to preferentially engage one pathway over another (9)(10)(11)(12). Stabilization of such distinct conformations by different ligands or mutant forms of ligands or receptors thus enables the preferential activation of specific subsets of effectors and downstream signaling pathways.…”

Section: Introductionmentioning

confidence: 99%

Functional selectivity profiling of the angiotensin II type 1 receptor using pathway-wide BRET signaling sensors

et al. 2018

Self Cite

View full text Add to dashboard Cite

G protein–coupled receptors (GPCRs) are important therapeutic targets that exhibit functional selectivity (biased signaling), in which different ligands or receptor variants elicit distinct downstream signaling. Understanding all the signaling events and biases that contribute to both the beneficial and adverse effects of GPCR stimulation by given ligands is important for drug discovery. Here, we report the design, validation, and use of pathway-selective bioluminescence resonance energy transfer (BRET) biosensors that monitor the engagement and activation of signaling effectors downstream of G proteins, including protein kinase C (PKC), phospholipase C (PLC), p63RhoGEF, and Rho. Combined with G protein and β-arrestin BRET biosensors, our sensors enabled real-time monitoring of GPCR signaling at different levels in downstream pathways in both native and engineered cells. Profiling of the responses to 14 angiotensin II (AngII) type 1 receptor (AT1R) ligands enabled the clustering of compounds into different subfamilies of biased ligands and showed that, in addition to the previously reported functional selectivity between Gαq and β-arrestin, there are also biases among G protein subtypes. We also demonstrated that biases observed at the receptor and G protein levels propagated to downstream signaling pathways and that these biases could occur through the engagement of different G proteins to activate a common effector. We also used these tools to determine how naturally occurring AT1R variants affected signaling bias. This suite of BRET biosensors provides a useful resource for fingerprinting biased ligands and mutant receptors and for dissecting functional selectivity at various levels of GPCR signaling.

show abstract

Evolutionary action and structural basis of the allosteric switch controlling β2AR functional selectivity

Cited by 67 publications

References 68 publications

Characterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation

Characterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation

Mammalian Eps15 homology domain 1 potentiates angiogenesis of non-small cell lung cancer by regulating β2AR signaling

Functional selectivity profiling of the angiotensin II type 1 receptor using pathway-wide BRET signaling sensors

Contact Info

Product

Resources

About