Elucidation of Binding Sites of Dual Antagonists in the Human Chemokine Receptors CCR2 and CCR5

Hall, Spencer E.; Mao, Allen; Nicolaidou, Vicky; Finelli, Mattéa J.; Wise, Emma L.; Nedjai, Belinda; Kanjanapangka, Julie; Harirchian, Paymann; Chen, Deborah; Selchau, Victor; Ribeiro, Sofia; Schyler, Sabine; Pease, James E.; Horuk, Richard; Vaidehi, Nagarajan

doi:10.1124/mol.108.053470

Cited by 55 publications

(67 citation statements)

References 38 publications

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“…binding pocket of CCR2. RS504393 and Teijin interact with the highly conserved glutamic acid residue E291, most likely via their basic nitrogen atom (Mirzadegan et al, 2000;Hall et al, 2009). For BMS22, the adjacent T292 was found to be important for binding (Cherney et al, 2008), indicating that it shares the same binding pocket as RS504393 and Teijin.…”

Section: Discussionmentioning

confidence: 99%

“…dx.doi.org/10.1124/mol.113.086850. their binding sites can only partly overlap. For CCR2, several mutagenesis studies have provided evidence for the binding of small-molecule antagonists in the major and minor pocket (Mirzadegan et al, 2000;Berkhout et al, 2003;Hall et al, 2009). These ligands often contain a positively charged basic nitrogen that interacts with the conserved negatively charged glutamic acid residue (E291) in TM7, which is directly located between the major and minor binding pocket (Rosenkilde and Schwartz, 2006).…”

Section: Introductionmentioning

confidence: 99%

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Multiple Binding Sites for Small-Molecule Antagonists at the CC Chemokine Receptor 2

et al. 2013

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The chemokine receptor CCR2 is a G protein-coupled receptor that is activated primarily by the endogenous CC chemokine ligand 2 (CCL2). Many different small-molecule antagonists have been developed to inhibit this receptor, as it is involved in a variety of diseases characterized by chronic inflammation. Unfortunately, all these antagonists lack clinical efficacy, and therefore a better understanding of their mechanism of action is warranted. In this study, we examined the pharmacological properties of smallmolecule CCR2 antagonists in radioligand binding and functional assays. Six structurally different antagonists were selected for this study, all of which displaced the endogenous agonist 125

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multiple Binding Sites for Small-Molecule Antagonists at the CC Chemokine Receptor 2

et al. 2013

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“…The chimeric receptor CCR5-CCR2 all was previously described (Thiele et al, 2011), whereas CCR2-CCR5 C-term and CCR5-CCR2 C-term were designed in-house and cloned using polymerase chain reaction overlap extension technique (Piscataway, NJ). pcDNA3.11 plasmid containing the WT CCR2 with a 3Â hemagglutinin (HA) epitope tag at the N terminus was kindly provided by James Pease (Imperial College London, UK) (Hall et al, 2009). The promiscuous G protein Ga D6qi4myr (G qi4myr ) was kindly provided by Evi Kostenis (University of Bonn, Germany).…”

Section: Methodsmentioning

confidence: 99%

Discovery and Mapping of an Intracellular Antagonist Binding Site at the Chemokine Receptor CCR2

et al. 2014

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The chemokine receptor CCR2 is a G protein-coupled receptor that is involved in many diseases characterized by chronic inflammation, and therefore a large variety of CCR2 small molecule antagonists has been developed. On the basis of their chemical structures these antagonists can roughly be divided into two groups with most likely two topographically distinct binding sites. The aim of the current study was to identify the binding site of one such group of ligands, exemplified by three allosteric antagonists, CCR2-RA-[R], JNJ-27141491, and SD-24. We first used a chimeric CCR2/CCR5 receptor approach to obtain insight into the binding site of the allosteric antagonists and additionally introduced eight single point mutations in CCR2 to further characterize the putative binding pocket. All constructs were studied in radioligand binding and/or functional IP turnover assays, providing evidence for an intracellular binding site for CCR2-RA-[R], JNJ-27141491, and SD-24. For CCR2-RA-[R] the most important residues for binding were found to be the highly conserved tyrosine Y 7.53 and phenylalanine F 8.50 of the NPxxYx (5,6) F motif, as well as V 6.36 at the bottom of TM-VI and K 8.49 in helix-VIII. These findings demonstrate for the first time the presence of an allosteric intracellular binding site for CCR2 antagonists. This contributes to an increased understanding of the interactions of diverse ligands at CCR2 and may allow for a more rational design of future allosteric antagonists.

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“…The selection of the residue Glu291 was grounded from the reported results of sitedirected mutagenesis studies. 18,19 Several residues in the upper helices region of CCR2 have been shown to participate in the binding of small molecule ligands. Among those residues, the interaction between the acidic side chain Glu291 in TM helix VII and the basic amine of the ligand has been considered critical in CCR2, as well as in other chemokine receptor family members.…”

Section: Notesmentioning

confidence: 99%

“…18 However, Hall et al reported the model in which the amide hydrogen in TAK-779 interacts with Glu291. 19 In this study, Tyr49 showed the hydrogen bonding with the oxygen in the tetrahydro-pyranyl moiety in TAK-779 binding site model. This interaction was also observed in the model by Hall et al Table 2 shows Tyr49 has significant influence on Table 2.…”

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confidence: 99%

Prediction of Binding Mode between Chemokine Receptor CCR2 and Its Known Antagonists using Ligand Supported Homology Modeling

Kim

Lim²,

Lee³

et al. 2012

Bulletin of the Korean Chemical Society

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Chemokines play a crucial role in the trafficking of leukocytes in the body through the binding to their related receptors. Chemokine Receptor 2 (CCR2) belongs to GProtein Coupled Receptor (GPCR) family and expressed in monocytes, immature dendritic cells, activated T lymphocytes, basophils, and endothelial and vascular smooth-muscle cells. CCR2 binds several chemokines; CCL2, CCL7, CCL8, and CCL13.1 CCR2 and its ligand have been implicated in the pathophysiology of a number of diseases, including rheumatoid arthritis, multiple sclerosis, atherosclerosis, organ transplant rejection, and insulin resistance.2 Knowledge of the structural basis on CCR2-ligand interaction could help facilitate the design of novel CCR2 antagonists. We modeled and predicted the binding sites of widely known CCR2 antagonists, TAK-779 3 and Teijin-lead 4 ( Fig. 1), using ligand supported homology modeling method.Homology modeling predicts the three-dimensional structure of a given protein sequence based on its alignment to reference proteins of known three-dimensional structure (socalled templates). It is known as the most successful technique for predicting the three-dimensional protein structure. 5In conventional homology modeling method, the ligand molecules are not considered in the process of protein model building. After building the protein model, the ligand molecules are docked into the protein model. These approaches often showed the consequence that residue side chains involved in ligand binding are inappropriately modeled. The time-consuming re-modeling of the binding site or docking is required to generate sensible side-chain and ligand orientations. In the case of GPCR, the conventional homology modeling approaches for obtaining the binding site models are more challenging because the number of the available templates is very limited and the sequence identity to the templates is very low. The available templates for GPCR are the solved three-dimensional structures of bovine rhodopsin, 6 β 2 -adrenergic receptor (β 2 AR), 7 turkey β 1 -adrenergic receptor, 8 and human adenosine A 2A receptor. 9The homology modeling from those limited number of templates may result in uncertainty not only regarding backbone positioning but also about side chain conformations. One approach to solve this is to optimize the side chains through mechanics and/or dynamics in an empty pocket; however, this method may be inaccurate due to possible disruption of the binding site. Another approach is to use dynamics and ligand data as a restraint. This method also has a possibility of poor quality. 10 Thus, the development of modified homology modeling methods for constructing a plausible ligand binding mode is an important theme in GPCR-ligand modeling research.The incorporation of ligand information from the early stage of homology modeling has been studied as an alternative strategy to the conventional homology modeling. This approach was first developed and termed as ligand supported homology modeling by Klebe and coworkers.11 Recently, the improved...

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Elucidation of Binding Sites of Dual Antagonists in the Human Chemokine Receptors CCR2 and CCR5

Abstract: Design of dual antagonists for the chemokine receptors CCR2 and CCR5 will be greatly facilitated by knowledge of the structural differences of their binding sites. Thus, we computationally predicted the binding site of the dual CCR2/CCR5 antagonist N

Cited by 55 publications

References 38 publications

Multiple Binding Sites for Small-Molecule Antagonists at the CC Chemokine Receptor 2

Multiple Binding Sites for Small-Molecule Antagonists at the CC Chemokine Receptor 2

Discovery and Mapping of an Intracellular Antagonist Binding Site at the Chemokine Receptor CCR2

Prediction of Binding Mode between Chemokine Receptor CCR2 and Its Known Antagonists using Ligand Supported Homology Modeling

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