Molecular Determinants of CGS21680 Binding to the Human Adenosine A<sub>2A</sub> Receptor

Lebon, Guillaume; Edwards, Patricia C.; Leslie, Andrew G. W.; Tate, Christopher G.

doi:10.1124/mol.114.097360

Cited by 133 publications

(165 citation statements)

References 47 publications

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“…S3). According to the density map and refinement results, the conformation in which the side chain of Phe168 projects into the receptor core and makes π-π interaction with the ligand seems to be the dominant conformation at occupancy of 0.7, which is consistent with its conformation in all other reported A 2A R structures (7)(8)(9)(10)(11)(12)(13)(14). Further discussion of the binding pocket will be based on this single conformation.…”

Section: Significancesupporting

confidence: 62%

“…To understand the structureactivity relationship around the chemical series for the A 2A R and facilitate further optimization, we sought to obtain the crystal structure of the A 2A R in complex with Cmpd-1. Although a number of crystal structures of A 2A R bound to various antagonists and agonists have been reported (7)(8)(9)(10)(11)(12)(13)(14) using lipidic cubic phase (LCP) methods (15,16), we were unable to obtain high-quality crystals of A 2A R bound to Cmpd-1. As an alternative approach we obtained a 3.5-Å resolution crystal structure of A 2A R in complex with Cmpd-1 by using a BRIL fusion construct and performing the crystallization in vapor-phase diffusion in the detergent lauryl maltose neopentyl glycol (LMNG).…”

mentioning

confidence: 87%

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Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Sun

Bachhawat

Chu

et al. 2017

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

122

100

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The adenosine A 2A receptor (A 2A R) has long been implicated in cardiovascular disorders. As more selective A 2A R ligands are being identified, its roles in other disorders, such as Parkinson's disease, are starting to emerge, and A 2A R antagonists are important drug candidates for nondopaminergic anti-Parkinson treatment. Here we report the crystal structure of A 2A receptor bound to compound 1 (Cmpd-1), a novel A 2A R/N-methyl D-aspartate receptor subtype 2B (NR2B) dual antagonist and potential anti-Parkinson candidate compound, at 3.5 Å resolution. The A 2A receptor with a cytochrome b562-RIL (BRIL) fusion (A 2A R-BRIL) in the intracellular loop 3 (ICL3) was crystallized in detergent micelles using vapor-phase diffusion. Whereas A 2A R-BRIL bound to the antagonist ZM241385 has previously been crystallized in lipidic cubic phase (LCP), structural differences in the Cmpd-1-bound A 2A R-BRIL prevented formation of the lattice observed with the ZM241385-bound receptor. The crystals grew with a type II crystal lattice in contrast to the typical type I packing seen from membrane protein structures crystallized in LCP. Cmpd-1 binds in a position that overlaps with the native ligand adenosine, but its methoxyphenyl group extends to an exosite not previously observed in other A 2A R structures. Structural analysis revealed that Cmpd-1 binding results in the unique conformations of two tyrosine residues, Tyr9 1.35 and Tyr271 7.36 , which are critical for the formation of the exosite. The structure reveals insights into antagonist binding that are not observed in other A 2A R structures, highlighting flexibility in the binding pocket that may facilitate the development of A 2A R-selective compounds for the treatment of Parkinson's disease.T he adenosine A 2A receptor (A 2A R) is one of the four subtypes of adenosine receptors (A 1 R, A 2A R, A 2B R, A 3 R). As a member of the G protein-coupled receptor (GPCR) family, the A 2A receptor couples to stimulatory G protein G s and elevates intracellular cAMP upon activation by endogenous adenosine. The A 2A R has been an attractive drug target due to its role in cardiovascular and immune system function, as well as in the central nervous system as a potential therapeutic target for Parkinson's disease (PD) (1-4). PD is a neurodegenerative disease that affects more than 1% of the population over 65 years old. Currently, major treatments target the restoration of dopamine signaling, which is impaired in PD patients, by dopamine-replacing agents. Although these treatments effectively address PD-related motor disturbances, the long-term use of dopamine-replacing agents is associated with the development of motor complications; therefore, there is a need for nondopaminergic drugs (2). It is known that A 2A R signaling regulates dopaminergic neurotransmission, and A 2A R antagonists have been shown to enhance D2 dopamine receptor signaling, which has been found to improve PD symptoms in animal models and patients, without the side effects common to dopaminereplacing agents, such...

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Section: Significancesupporting

confidence: 62%

mentioning

confidence: 87%

Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Sun

Bachhawat

Chu

et al. 2017

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

122

100

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“…This constant did not change in the presence of several A 2A R antagonists, such as caffeine or SCH 58261 (Table 1). Due to the fact that both agonists and antagonists of A 2A R bind and compete for the same orthosteric site according to crystallographic studies [38][39][40][41], agonist-induced changes seen in the dissociation constant of the antagonist occur because both are bound simultaneously to different protomers in an A 2A R-A 2A R homodimer [15,21]. Likewise, dissociation experiments of [ 3 H]YM-09151-2 in the same striatal preparations showed that the D 2 R agonist quinpirole, but not the antagonist raclopride, significantly modified the dissociation rate of the radiolabelled antagonist [ 3 H]YM-09151-2 (Table 1), indicating the formation of a hybrid species with both agonist and antagonist simultaneously binding to the D 2 R-D 2 R homodimer.…”

Section: Homodimeric Nature Of Both a 2a R And D 2 Rmentioning

confidence: 99%

Evidence for the heterotetrameric structure of the adenosine A2A–dopamine D2 receptor complex

Casadó-Anguera

Bonaventura

Moreno

et al. 2016

Biochemical Society Transactions

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Heteromers of G-protein-coupled receptors (GPCRs) have emerged as potential novel targets for drug development. Accumulating evidence indicates that GPCRs can form homodimers and heteromers, with homodimers being the predominant species and oligomeric receptors being formed as multiples of dimers. Recently, heterotetrameric structures have been proposed for dopamine D1receptor (D1R)-dopamine D3receptor (D3R) and adenosine A2Areceptor (A2AR)-dopamine D2receptor (D2R) heteromers. The structural model proposed for these complexes is a heteromer constituted by two receptor homodimers. The existence of GPCR homodimers and heteromers provides a structural basis for inter-protomer allosteric mechanisms that might account for a multiplicity of unique pharmacological properties. In this review, we focus on the A2AR-D2R heterotetramer as an example of an oligomeric structure that is key in the modulation of striatal neuronal function. We also review the interfaces involved in this and other recently reported heteromers of GPCRs. Furthermore, we discuss several published studies showing theex vivoexpression of A2AR-D2R heteromers. The ability of A2AR agonists to decrease the affinity of D2R agonists has been reported and, on the basis of this interaction, A2AR antagonists have been proposed as potential drugs for the treatment of Parkinson's disease. The heterotetrameric structure of the A2AR-D2R complex offers a novel model that can provide new clues about how to adjust the drug dosage to the expected levels of endogenous adenosine.

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“…3), indicating formation of a hybrid species with both agonist and antagonist simultaneously binding to the dimer. Therefore, only the agonist can exert an allosteric modulation of the labeled antagonist when both are occupying orthosteric sites in an A 2A R oligomer, because the four ligands-caffeine, ZM 241385, SCH 58261, and CGS 21680-all bind and compete for the same orthosteric site (19)(20)(21). This implies a different conformation of the A 2A R homodimer when occupied simultaneously with an agonist and an antagonist compared with when occupied with two antagonists.…”

mentioning

confidence: 99%

Allosteric interactions between agonists and antagonists within the adenosine A _2A receptor-dopamine D ₂ receptor heterotetramer

Bonaventura

Navarro

Casadó-Anguera

et al. 2015

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

137

220

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Adenosine A 2A receptor (A 2A R)-dopamine D 2 receptor (D 2 R) heteromers are key modulators of striatal neuronal function. It has been suggested that the psychostimulant effects of caffeine depend on its ability to block an allosteric modulation within the A 2A R-D 2 R heteromer, by which adenosine decreases the affinity and intrinsic efficacy of dopamine at the D 2 R. We describe novel unsuspected allosteric mechanisms within the heteromer by which not only A 2A R agonists, but also A 2A R antagonists, decrease the affinity and intrinsic efficacy of D 2 R agonists and the affinity of D 2 R antagonists. Strikingly, these allosteric modulations disappear on agonist and antagonist coadministration. This can be explained by a model that considers A 2A R-D 2 R heteromers as heterotetramers, constituted by A 2A R and D 2 R homodimers, as demonstrated by experiments with bioluminescence resonance energy transfer and bimolecular fluorescence and bioluminescence com- Most evidence indicates that G protein-coupled receptors (GPCRs) form homodimers and heteromers. Homodimers seem to be a predominant species, and oligomeric entities can be viewed as multiples of dimers (1). It has been proposed that GPCR heteromers are constituted mainly by heteromers of homodimers (1, 2). Allosteric mechanisms determine a multiplicity of unique pharmacologic properties of GPCR homodimers and heteromers (1, 3). First, binding of a ligand to one of the receptors in the heteromer can modify the affinity of ligands for the other receptor (1, 3, 4). The most widely reproduced allosteric modulation of ligand-binding properties in a GPCR heteromer is the ability of adenosine A 2A receptor (A 2A R) agonists to decrease the affinity of dopamine D 2 receptor (D 2 R) agonists in the A 2A R-D 2 R heteromer (5). A 2A R-D 2 R heteromers have been revealed both in transfected cells (6, 7), striatal neurons in culture (6,8) and in situ, in mammalian striatum (9, 10), where they play an important role in the modulation of GABAergic striatopallidal neuronal function (9, 11).In addition to ligand-binding properties, unique properties for each GPCR oligomer emerge in relation to the varying intrinsic efficacy of ligands for different signaling pathways (1-3). Intrinsic efficacy refers to the power of the agonist to induce a functional response, independent of its affinity for the receptor. Thus, allosteric modulation of an agonist can potentially involve changes in affinity and/or intrinsic efficacy (1, 3). This principle can be observed in the A 2A R-D 2 R heteromer, where a decrease in D 2 R agonist affinity cannot alone explain the ability of an A 2A R agonist to abolish the decreased excitability of GABAergic striatopallidal neurons induced by high concentration of a D 2 R agonist (9), which should overcome the decrease in affinity. Furthermore, a differential effect of allosteric modulations of different agonist-mediated signaling responses (i.e., functional selectivity) can occur within GPCR heteromers (1, 2, 8 It has been hypothesized that the allos...

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Molecular Determinants of CGS21680 Binding to the Human Adenosine A_2A Receptor

Abstract: The adenosine A 2A receptor (A 2A R) plays a key role in transmembrane signaling mediated by the endogenous agonist adenosine. Here, we describe the crystal structure of human A 2A

Cited by 133 publications

References 47 publications

Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Evidence for the heterotetrameric structure of the adenosine A2A–dopamine D2 receptor complex

Allosteric interactions between agonists and antagonists within the adenosine A _2A receptor-dopamine D ₂ receptor heterotetramer

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Molecular Determinants of CGS21680 Binding to the Human Adenosine A2A Receptor

Abstract: The adenosine A 2A receptor (A 2A R) plays a key role in transmembrane signaling mediated by the endogenous agonist adenosine. Here, we describe the crystal structure of human A 2A

Cited by 133 publications

References 47 publications

Crystal structure of the adenosine A 2A receptor bound to an antagonist reveals a potential allosteric pocket

Crystal structure of the adenosine A 2A receptor bound to an antagonist reveals a potential allosteric pocket

Evidence for the heterotetrameric structure of the adenosine A2A–dopamine D2 receptor complex

Allosteric interactions between agonists and antagonists within the adenosine A 2A receptor-dopamine D 2 receptor heterotetramer

Contact Info

Product

Resources

About

Molecular Determinants of CGS21680 Binding to the Human Adenosine A_2A Receptor

Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Crystal structure of the adenosine A _2A receptor bound to an antagonist reveals a potential allosteric pocket

Allosteric interactions between agonists and antagonists within the adenosine A _2A receptor-dopamine D ₂ receptor heterotetramer