Intrinsic bias at non-canonical, β-arrestin-coupled seven transmembrane receptors

Pandey, Shubhi; Kumari, Punita; Baidya, Mithu; Kise, Ryoji; Cao, Yuan; Dwivedi-Agnihotri, Hemlata; Banerjee, Rita; Li, Xaria X.; Cui, Cedric S.; Lee, John D.; Kawakami, Kouki; Maharana, Jagannath; Ranjan, Amit; Chaturvedi, Madhu; Jhingan, Gagan Deep; Laporte, Stéphane A.; Woodruff, Trent M.; Inoue, Asuka; Shukla, Arun K.

doi:10.1016/j.molcel.2021.09.007

Cited by 80 publications

(89 citation statements)

References 73 publications

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“…While this manuscript was under revision, another group published three independently created ΔGRK knockout cell lines (ΔGRK2/3, ΔGRK5/6 and ΔGRK2/3/5/6) 69 , which correspond to our GRK family knockout cell lines and ΔQ-GRK. The only common receptor among both manuscripts is the C5aR1.…”

Section: Discussionmentioning

confidence: 99%

GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation

et al. 2022

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G protein-coupled receptors (GPCRs) activate G proteins and undergo a complex regulation by interaction with GPCR kinases (GRKs) and the formation of receptor–arrestin complexes. However, the impact of individual GRKs on arrestin binding is not clear. We report the creation of eleven combinatorial HEK293 knockout cell clones lacking GRK2/3/5/6, including single, double, triple and the quadruple GRK knockout. Analysis of β-arrestin1/2 interactions for twelve GPCRs in our GRK knockout cells enables the differentiation of two main receptor subsets: GRK2/3-regulated and GRK2/3/5/6-regulated receptors. Furthermore, we identify GPCRs that interact with β-arrestins via the overexpression of specific GRKs even in the absence of agonists. Finally, using GRK knockout cells, PKC inhibitors and β-arrestin mutants, we present evidence for differential receptor–β-arrestin1/2 complex configurations mediated by selective engagement of kinases. We anticipate our GRK knockout platform to facilitate the elucidation of previously unappreciated details of GRK-specific GPCR regulation and β-arrestin complex formation.

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Section: Discussionmentioning

confidence: 99%

GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation

et al. 2022

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“…System bias includes a receptor's G protein or arrestin selectivity. For example, some GPCRs lack the inherent ability to elicit G protein coupling while exhibiting robust arrestin interaction (Meyrath et al, 2020;Pandey et al, 2021;Rajagopal et al, 2010).…”

Section: Ligand Bias Definition and Distinction From System Biasmentioning

confidence: 99%

“…System bias includes a receptor’s G protein or arrestin selectivity. For example, some GPCRs lack the inherent ability to elicit G protein coupling while exhibiting robust arrestin interaction ( Meyrath et al, 2020 ; Pandey et al, 2021 ; Rajagopal et al, 2010 ). Furthermore, system bias depends on the different levels of receptor constitutive activity, and intracellular intra- or inter-pathway feedbacks.…”

Section: Definitions Of Pathways and Of Bias Typesmentioning

confidence: 99%

Community guidelines for GPCR ligand bias: IUPHAR review 32

Kolb

Kenakin

Alexander

et al. 2022

British J Pharmacology

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GPCRs modulate a plethora of physiological processes and mediate the effects of one-third of FDA-approved drugs. Depending on which ligand activates a receptor, it can engage different intracellular transducers. This 'biased signalling' paradigm requires that we now characterize physiological signalling not just by receptors but by ligand-receptor pairs. Ligands eliciting biased signalling may constitute better drugs with higher efficacy and fewer adverse effects. However, ligand bias is very complex, making reproducibility and description challenging. Here, we provide guidelines and terminology for any scientists to design and report ligand bias experiments.The guidelines will aid consistency and clarity, as the basic receptor research and drug discovery communities continue to advance our understanding and exploitation of ligand bias. Scientific insight, biosensors, and analytical methods are still evolving and should benefit from and contribute to the implementation of the guidelines, together improving translation from in vitro to disease-relevant in vivo models. | INTRODUCTIONThe $800 human GPCRs transduce sensory inputs and systemic signals into appropriate cellular responses in numerous physiological processes. They recognize a vast diversity of signals ranging from photons, tastants and odours to ions, neurotransmitters, hormones, and cytokines (Harding et al., 2021;Wacker, Stevens, & Roth, 2017).Even though GPCRs represent the primary target of 34% of FDAapproved drugs, more than 220 non-olfactory GPCRs have disease associations which are as yet untapped in clinical research (Hauser, Attwood, Rask-Andersen, Schioth, & Gloriam, 2017;Sriram & Insel, 2018). Despite the diversity of extracellular ligands and physiological roles of GPCRs, these cell surface receptors share a conserved molecular fold and intracellular transducers. Agonist binding stabilizes active conformations of the receptor, facilitating the binding of one or more cytosolic transducer proteins. These include the heterotrimeric G proteins consisting of α, β and γ subunits that dissociate to α and βγ upon activation by the receptor. G proteins comprise 16 distinct α subunits and are divided into four families based on homology and associated downstream signalling pathways: G s (G s and

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“…C5a and its interaction with C5aR1 is associated with the induction of neutrophil mobilization and recruitment. , As the goal of this study was to design a functionally similar agonist to C5a for C5aR1, the ability of BM213 and BM221 to induce neutrophil mobilization in mice was assessed via a polymorphonuclear leucocyte (PMN) mobilization assay. A dosage of 50 μg kg –1 for both BM213 and BM221, administered via intravenous injection, was sufficient to induce neutrophil mobilization comparable to the administration of mouse C5a at the same dosage (Figure B).…”

Section: Resultsmentioning

confidence: 99%

Development of Potent and Selective Agonists for Complement C5a Receptor 1 with In Vivo Activity

Gorman

Lee

et al. 2021

J. Med. Chem.

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The anaphylatoxin C5a is a complement peptide associated with immune-related disorders. C5a binds with equal potency to two GPCRs, C5aR1 and C5aR2. Multiple C5a peptide agonists have been developed to interrogate the C5a receptor function but none show selectivity for C5aR1. To address these limitations, we developed potent and stable peptide C5aR1 agonists that display no C5aR2 activity and over 1000-fold selectivity for C5aR1 over C3aR. This includes BM213, which induces C5aR1-mediated calcium mobilization and pERK1/2 signaling but not β-arrestin recruitment, and BM221, which exhibits no signaling bias. Both ligands are functionally similar to C5a in human macrophage cytokine release assays and in a murine in vivo neutrophil mobilization assay. BM213 showed antitumor activity in a mouse model of mammary carcinoma. We anticipate that these C5aR1-selective agonists will be useful research tools to investigate C5aR1 function.

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Intrinsic bias at non-canonical, β-arrestin-coupled seven transmembrane receptors

Cited by 80 publications

References 73 publications

GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation

GPCR kinase knockout cells reveal the impact of individual GRKs on arrestin binding and GPCR regulation

Community guidelines for GPCR ligand bias: IUPHAR review 32

Development of Potent and Selective Agonists for Complement C5a Receptor 1 with In Vivo Activity

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