Selectivity Landscape of 100 Therapeutically Relevant GPCR Profiled by an Effector Translocation-Based BRET Platform

Avet, Charlotte; Mancini, Arturo; Breton, Billy; Gouill, Christian Le; Hauser, Alexander S.; Normand, Claire; Kobayashi, Hiroyuki; Gross, Florence; Hogue, Mireille; Lukasheva, Viktoriya; Morissette, Sandra; Fauman, Eric B.; Fortin, Jean‐Philippe; Schann, Stephane; Leroy, Xavier; Gloriam, David E.; Bouvier, Michel

doi:10.2139/ssrn.3586569

Cited by 26 publications

(39 citation statements)

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“…Unexpectedly, among the investigated modulators, 100 ng/mL CTX had the strongest effect on E max at the hH 1 R lowering the value to 23 ± 4.9% (Figure 6A), suggesting that the Gα s protein is involved in the hH 1 R mediated DMR signal. Indeed, it has been shown that the hH 1 R can functionally interact with the Gα s protein in HEK cells overexpressing both the receptor and the Gα s protein [60,91]. The inhibition of Gα s pathway led to a significant increase in the pEC 50 value (7.87 ± 0.19; Figure 6B).…”

Section: • Hh2rmentioning

confidence: 92%

“…We conclude that Gα q/11 , Gα s , and Gα i/o played a minor role in the generation of the HIS DMR signal in HEK hH 2 R cells, and that Gα 12/13 proteins must have been involved. It has already been described in the literature that the hH 2 R is capable to interact with the Gα 12/13 protein [59,60], however, it was unexpected that the involvement of Gα 12/13 would exceed the contribution of Gα q/11 , Gα s , and Gα i/o .…”

Section: Pharmacological Versus Molecular Biological Approach To Silence Gα Proteinmentioning

confidence: 94%

“…The optical traces (representations in Figure 2A, C) were converted to CRCs by calculating the AUC 60 and plotting these values against the logarithmic concentrations of a compound (Figure 4A). The determined pEC 50 and E max values are summarized in Table 2.…”

Section: Conversion Of the Dmr Responses To Concentration-response-curves (Crcs)mentioning

confidence: 99%

“…They transmit their signals predominantly via three classes of G proteins: H 1 via Gα q/11 , H 2 via Gα s and H 3 + H 4 via Gα i/o (Figure 1B). However, for the H 1 and H 2 receptors, evidence is emerging for promiscuous activation of Gα proteins [59][60][61]. By contrast, less information is available on the involvement of non-canonical G protein subunits in the signal transduction processes of the H 3,4 Rs.…”

Section: Introductionmentioning

confidence: 99%

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Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors

Seibel-Ehlert

Plank

Inoue

et al. 2021

IJMS

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G protein activation represents an early key event in the complex GPCR signal transduction process and is usually studied by label-dependent methods targeting specific molecular events. However, the constrained environment of such “invasive” techniques could interfere with biological processes. Although histamine receptors (HRs) represent (evolving) drug targets, their signal transduction is not fully understood. To address this issue, we established a non-invasive dynamic mass redistribution (DMR) assay for the human H1–4Rs expressed in HEK cells, showing excellent signal-to-background ratios above 100 for histamine (HIS) and higher than 24 for inverse agonists with pEC50 values consistent with literature. Taking advantage of the integrative nature of the DMR assay, the involvement of endogenous Gαq/11, Gαs, Gα12/13 and Gβγ proteins was explored, pursuing a two-pronged approach, namely that of classical pharmacology (G protein modulators) and that of molecular biology (Gα knock-out HEK cells). We showed that signal transduction of hH1–4Rs occurred mainly, but not exclusively, via their canonical Gα proteins. For example, in addition to Gαi/o, the Gαq/11 protein was proven to contribute to the DMR response of hH3,4Rs. Moreover, the Gα12/13 was identified to be involved in the hH2R mediated signaling pathway. These results are considered as a basis for future investigations on the (patho)physiological role and the pharmacological potential of H1–4Rs.

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Section: • Hh2rmentioning

confidence: 92%

Section: Pharmacological Versus Molecular Biological Approach To Silence Gα Proteinmentioning

confidence: 94%

Section: Conversion Of the Dmr Responses To Concentration-response-curves (Crcs)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors

Seibel-Ehlert

Plank

Inoue

et al. 2021

IJMS

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“…Here, we aimed to develop new tools to monitor ERM activation. We elected to develop ERM biosensors based on the principle of bioluminescence resonance energy transfer (BRET) (Angers et al, 2000;Xu et al, 1999) and enhanced-bystander BRET (ebBRET) (Namkung et al, 2016), since such sensors are easy to use in living cells and are compatible with high-throughput screening (Avet et al, 2020;Benredjem et al, 2019;Namkung et al, 2018;Schönegge et al, 2017). BRET is similar to fluorescence resonance energy transfer (FRET) except that the energy donor is a luminescent and not a fluorescent molecule.…”

Section: Introductionmentioning

confidence: 99%

Development of conformational BRET biosensors that monitor ezrin, radixin and moesin activation in real time

Leguay

Decelle

et al. 2021

Journal of Cell Science

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Ezrin, Radixin and Moesin compose the family of ERM proteins. They link actin filaments and microtubules to the plasma membrane to control signaling and cell morphogenesis. Importantly, their activity promotes invasive properties of metastatic cells from different cancer origins. Therefore, a precise understanding of how these proteins are regulated is important for the understanding of the mechanism controlling cell shape as well as providing new opportunities for the development of innovative cancer therapies. Here, we developed and characterized novel BRET-based conformational biosensors, compatible with high throughput screening, that monitor Ezrin, Radixin or Moesin individual activation in living cells. We showed that these biosensors faithfully monitor ERM activation and can be used to quantify the impact of small molecules, mutation of regulatory amino acids or depletion of upstream regulators on their activity. The use of these biosensors allowed us to characterize the activation process of ERMs that involves a pool of closed-inactive ERMs stably associated with the plasma membrane. Upon stimulation, we discovered that this pool serves as a cortical reserve that is rapidly activated before the recruitment of cytoplasmic ERMs.

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Personalized Medicine Through GPCR Pharmacogenomics

Hauser

2022

Comprehensive Pharmacology

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Selectivity Landscape of 100 Therapeutically Relevant GPCR Profiled by an Effector Translocation-Based BRET Platform

Cited by 26 publications

References 0 publications

Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors

Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors

Development of conformational BRET biosensors that monitor ezrin, radixin and moesin activation in real time

Personalized Medicine Through GPCR Pharmacogenomics

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