Impact of GPCR Structures on Drug Discovery

Congreve, Miles; Graaf, Chris de; Swain, Nigel A.; Tate, Christopher G.

doi:10.1016/j.cell.2020.03.003

Cited by 260 publications

(249 citation statements)

References 103 publications

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“…G protein‐coupled receptors (GPCRs) remain important targets for drug discovery, and current approaches to GPCR drug discovery are structure‐based [1] . Many well‐established drugs that act at GPCRs, either directly at the receptor, or indirectly by modulating the concentration of their endogenous GPCR ligands, are clearly among the most highly prescribed pharmaceuticals for diverse therapeutic areas [2] .…”

Section: Introductionmentioning

confidence: 99%

“…The drugs introduced in previous decades that act at GPCRs were found mainly by empirical methods of probing structure–activity relationships (SAR), often inspired by natural products. However, modern ligand discovery for GPCRs is largely structure‐based, and knowledge of GPCR structures is rapidly accruing, beginning with the structures of bovine rhodopsin (2000) and later (2007) the β 2 ‐adrenergic receptor [1,5] . Hundreds of new structures determined in recent years have greatly aided this effort.…”

Section: Introductionmentioning

confidence: 99%

“…Table 1 shows current approaches used for the study of GPCRs, encompassing both biophysical approaches, novel assay methods, and sophisticated ligand design. Biophysical methods used to study GPCR structures include X‐ray crystallography, X‐ray free‐electron laser (XFEL), cryogenic electron microscopy (cryo‐EM), and NMR [7] . Also, advances in the computational modeling of membrane‐bound proteins like GPCRs have provided an interface for medicinal chemists to predict the effects of ligand structural changes on the GPCR binding and activation processes [8–10] .…”

Section: Introductionmentioning

confidence: 99%

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Purinergic Signaling: Impact of GPCR Structures on Rational Drug Design

Salmaso

2020

ChemMedChem

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The purinergic signaling system includes membrane‐bound receptors for extracellular purines and pyrimidines, and enzymes/transporters that regulate receptor activation by endogenous agonists. Receptors include: adenosine (A1, A2A, A2B, and A3) and P2Y (P2Y1, P2Y2, P2Y4, P2Y6, P2Y11, P2Y12, P2Y13, and P2Y14) receptors (all GPCRs), as well as P2X receptors (ion channels). Receptor activation, especially accompanying physiological stress or damage, creates a temporal sequence of signaling to counteract this stress and either mobilize (P2Rs) or suppress (ARs) immune responses. Thus, modulation of this large signaling family has broad potential for treating chronic diseases. Experimentally determined structures represent each of the three receptor families. We focus on selective purinergic agonists (A1, A3), antagonists (A3, P2Y14), and allosteric modulators (P2Y1, A3). Examples of applying structure‐based design, including the rational modification of known ligands, are presented for antithrombotic P2Y1R antagonists and anti‐inflammatory P2Y14R antagonists and A3AR agonists. A3AR agonists are a potential, nonaddictive treatment for chronic neuropathic pain.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Purinergic Signaling: Impact of GPCR Structures on Rational Drug Design

Salmaso

2020

ChemMedChem

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“…A detailed knowledge of GPCR structure is essential for effective drug discovery (Congreve, de Graaf et al 2020) and contributes to successful in silico ligand screening and ligand design (Lyu, Wang et al 2019). The process of obtaining a structure starts with purification of the target protein (Tate and Schertler 2009, Tate 2012, Milic and Veprintsev 2015.…”

Section: Introductionmentioning

confidence: 99%

ThermoFRET: A novel nanoscale G protein coupled receptor thermostability assay functional in crude solubilised membrane preparations

Tippett

Hoare

Miljuš

et al. 2020

Preprint

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AbstractSensitive protein stability assays for membrane proteins are crucial for developing purification protocols, for structural and biophysical characterisation and drug discovery. Here, we describe a novel high-throughput 384-well FRET-based thermostability methodology, ThermoFRET, allowing for the ultrasensitive determination of G protein coupled receptor (GPCR) stability. This method measures FRET between a terbium-cryptate labelled GPCR and BODIPY-FL-Cystine, a thiolreactive dye that reacts with cysteine residues exposed upon protein unfolding in response to thermal denaturation. ThermoFRET is functional in crude solubilised membrane preparations, without protein purification and can detect receptor stabilising ligands, making it ideally suited for orphan receptor screening.

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“…G protein coupled receptors (GPCRs) are a large family of membrane proteins that are important drug discovery targets (Hauser, Attwood et al 2017). Structural and biophysical studies of GPCRs have significant importance in modern drug discovery (Congreve, de Graaf et al 2020) but require receptor solubilisation from their native membrane environment and subsequent purification. Optimisation of receptor stability during this process is a key component to success (Tate 2010).…”

Section: Introductionmentioning

confidence: 99%

ThermoBRET: a ligand-engagement nanoscale thermostability assay applied to GPCRs

Hoare

Kaur

Dijon

et al. 2020

Preprint

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Sensitive assays to measure the thermostability of membrane proteins are important tools in protein purification optimisation and drug discovery. Here, we present a ThermoBRET method to quantify the relative thermostability of G protein coupled receptors (GPCRs), using cannabinoid receptor 2 (CB2) as an example. This method applies the principles of Bioluminescence Resonance Energy Transfer (BRET) between Nanoluciferase (Nluc) and a thiol-reactive fluorescent dye that covalently binds cysteines in the GPCR transmembrane domain, exposed by unfolding. We demonstrate that the melting point (Tm) of Nluc-fused GPCRs can be determined in non-purified detergent solubilised membrane preparations, revealing differences in thermostability for different detergent solubilising conditions and in the presence of stabilising ligands. In addition, we extended the range of the assay by developing the thermostable tsNLuc by incorporating mutations from the fragments of split-Nluc (Tm of 87 °C vs 59 °C). ThermoBRET allows the high-throughput determination of GPCR thermostability which will be useful for protein purification optimisation strategies and as part of a drug discovery screening platform.

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Impact of GPCR Structures on Drug Discovery

Cited by 260 publications

References 103 publications

Purinergic Signaling: Impact of GPCR Structures on Rational Drug Design

Purinergic Signaling: Impact of GPCR Structures on Rational Drug Design

ThermoFRET: A novel nanoscale G protein coupled receptor thermostability assay functional in crude solubilised membrane preparations

ThermoBRET: a ligand-engagement nanoscale thermostability assay applied to GPCRs

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