NMR analysis of GPCR conformational landscapes and dynamics

Casiraghi, Marina; Point, Elodie; Pozza, Alexandre; Moncoq, Karine; Banères, Jean-Louis; Catoire, Laurent J.

doi:10.1016/j.mce.2018.12.019

Cited by 31 publications

(21 citation statements)

References 101 publications

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“…Receptor crystal structures derived from X-ray diffraction data can bias conformations such that the ground state (R or LR) or fully active state (R* or LR*) is observed but the spectrum of graded or partial activity states between R/LR and LR/LR* is not. However, solution NMR spectroscopy studies are capable of detecting graded activity conformational states (10)(11)(12). Furthermore, to our knowledge, a direct quantitative assessment has yet to be reported on the relationship between graded ligand potency with functional efficacy and receptor conformation as predicted by the theoretical models of receptor activation.…”

Section: Significancementioning

confidence: 99%

Quantitative structural assessment of graded receptor agonism

Shang

Brust

Griffin

et al. 2019

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

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Ligand–receptor interactions, which are ubiquitous in physiology, are described by theoretical models of receptor pharmacology. Structural evidence for graded efficacy receptor conformations predicted by receptor theory has been limited but is critical to fully validate theoretical models. We applied quantitative structure–function approaches to characterize the effects of structurally similar and structurally diverse agonists on the conformational ensemble of nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ). For all ligands, agonist functional efficacy is correlated to a shift in the conformational ensemble equilibrium from a ground state toward an active state, which is detected by NMR spectroscopy but not observed in crystal structures. For the structurally similar ligands, ligand potency and affinity are also correlated to efficacy and conformation, indicating ligand residence times among related analogs may influence receptor conformation and function. Our results derived from quantitative graded activity–conformation correlations provide experimental evidence and a platform with which to extend and test theoretical models of receptor pharmacology to more accurately describe and predict ligand-dependent receptor activity.

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Section: Significancementioning

confidence: 99%

Quantitative structural assessment of graded receptor agonism

Shang

Brust

Griffin

et al. 2019

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

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“…Third, a single receptor can signal through several intracellular pathways, some of which could be preferentially activated by so-called "biased" ligands 6 . These three phenomena indicate that receptors are highly dynamic molecules and sample several active and inactive states stochastically (for review, see [7][8][9] ).…”

Section: Introductionmentioning

confidence: 99%

Sub-millisecond conformational dynamics of the A_2A adenosine receptor revealed by single-molecule FRET

Maslov

Volkov

Khorn

et al. 2020

Preprint

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The complex pharmacology of G-protein-coupled receptors (GPCRs) is defined by their multi-state conformational dynamics. Single-molecule Förster Resonance Energy Transfer (smFRET) is well-suited to quantify dynamics for individual protein molecules, however, its application to GPCRs is challenging; therefore, smFRET has been limited to studies of interreceptor interactions in cellular membranes and receptors in detergent environments. Here, we performed smFRET experiments on functionally active human A2A adenosine receptor (A2AAR) molecules embedded in freely diffusing lipid nanodiscs to study their intramolecular conformational dynamics. We propose a dynamic model of A2AAR activation that involves a slow (>2 ms) exchange between the active-like and inactive-like conformations in both apo and antagonist-bound A2AAR, explaining the receptor’s constitutive activity. For the agonist-bound A2AAR, we detected faster (390±80 μs) ligand efficacy-dependent dynamics. This work establishes a general smFRET platform for GPCR investigations that can potentially be used for drug screening and/or mechanism-of-action studies.

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“…It has been challenging to structurally observe the graded receptor activity conformations predicted by receptor theory. Although ligand-bound receptor crystal structures derived from X-ray diffraction data can bias conformations such that the ground or fully active state is observed but the spectrum of graded or partial activity states are not, solution NMR spectroscopy studies are capable of detecting graded activity conformational states (Boehr et al 2009, Kar et al 2010, Casiraghi et al 2019. Furthermore, to our knowledge, a direct quantitative assessment has yet to be reported on the relationship between graded ligand potency, efficacy, and receptor conformation as predicted by the two-state model of receptor activation (Leff 1995).…”

Section: Introductionmentioning

confidence: 99%

Quantitative Structural Assessment of Graded Receptor Agonism

Shang

Brust

Griffin

et al. 2019

Preprint

View full text Add to dashboard Cite

Ligand-receptor interactions, which are ubiquitous in physiology, are described by theoretical models of receptor pharmacology. Structural evidence for graded-efficacy receptor conformations predicted by receptor theory has been limited, but is critical to fully validate theoretical models. We applied quantitative structure-function approaches to characterize the effects of structurally similar and structurally diverse agonists on the conformational ensemble of nuclear receptor peroxisome proliferator-activated receptor gamma (PPARγ). For all ligands, agonist efficacy is correlated to a shift in the conformational ensemble equilibrium from a ground state towards an active state, which is detected by NMR spectroscopy but not observed in crystal structures. For the structurally similar ligands, ligand potency is also correlated to efficacy and conformation, indicating ligand residence times among related analogs can influence receptor conformation and function. Our results derived from quantitative graded activityconformation correlations provide new experimental evidence and a platform with which to extend and test theoretical models of receptor pharmacology to more accurately describe and predict ligand-dependent receptor activity.

show abstract

NMR analysis of GPCR conformational landscapes and dynamics

Cited by 31 publications

References 101 publications

Quantitative structural assessment of graded receptor agonism

Quantitative structural assessment of graded receptor agonism

Sub-millisecond conformational dynamics of the A_2A adenosine receptor revealed by single-molecule FRET

Quantitative Structural Assessment of Graded Receptor Agonism

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NMR analysis of GPCR conformational landscapes and dynamics

Cited by 31 publications

References 101 publications

Quantitative structural assessment of graded receptor agonism

Quantitative structural assessment of graded receptor agonism

Sub-millisecond conformational dynamics of the A2A adenosine receptor revealed by single-molecule FRET

Quantitative Structural Assessment of Graded Receptor Agonism

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Product

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Sub-millisecond conformational dynamics of the A_2A adenosine receptor revealed by single-molecule FRET