Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2

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Standard

Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2. / Zweemer, Annelien J M; Bunnik, Julia; Veenhuizen, Margo; Miraglia, Fabiana; Lenselink, Eelke B; Vilums, Maris; de Vries, Henk; Gibert, Arthur; Thiele, Stefanie; Rosenkilde, Mette M; IJzerman, Adriaan P; Heitman, Laura H.

I: Molecular Pharmacology, Bind 86, Nr. 4, 2014, s. 358-68.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Zweemer, AJM, Bunnik, J, Veenhuizen, M, Miraglia, F, Lenselink, EB, Vilums, M, de Vries, H, Gibert, A, Thiele, S, Rosenkilde, MM, IJzerman, AP & Heitman, LH 2014, 'Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2', Molecular Pharmacology, bind 86, nr. 4, s. 358-68. https://doi.org/10.1124/mol.114.093328

APA

Zweemer, A. J. M., Bunnik, J., Veenhuizen, M., Miraglia, F., Lenselink, E. B., Vilums, M., de Vries, H., Gibert, A., Thiele, S., Rosenkilde, M. M., IJzerman, A. P., & Heitman, L. H. (2014). Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2. Molecular Pharmacology, 86(4), 358-68. https://doi.org/10.1124/mol.114.093328

Vancouver

Zweemer AJM, Bunnik J, Veenhuizen M, Miraglia F, Lenselink EB, Vilums M o.a. Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2. Molecular Pharmacology. 2014;86(4):358-68. https://doi.org/10.1124/mol.114.093328

Author

Zweemer, Annelien J M ; Bunnik, Julia ; Veenhuizen, Margo ; Miraglia, Fabiana ; Lenselink, Eelke B ; Vilums, Maris ; de Vries, Henk ; Gibert, Arthur ; Thiele, Stefanie ; Rosenkilde, Mette M ; IJzerman, Adriaan P ; Heitman, Laura H. / Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2. I: Molecular Pharmacology. 2014 ; Bind 86, Nr. 4. s. 358-68.

Bibtex

@article{32d401e4234e453ea2d958f3668e8a07,
title = "Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2",
abstract = "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.",
keywords = "Allosteric Site, Amino Acid Motifs, Amino Acid Sequence, Animals, CHO Cells, COS Cells, Cell Line, Tumor, Cercopithecus aethiops, Cricetinae, Cricetulus, Humans, Imidazoles, Ligands, Molecular Sequence Data, Point Mutation, Protein Binding, Pyrrolidines, Receptors, CCR2, Sulfonamides",
author = "Zweemer, {Annelien J M} and Julia Bunnik and Margo Veenhuizen and Fabiana Miraglia and Lenselink, {Eelke B} and Maris Vilums and {de Vries}, Henk and Arthur Gibert and Stefanie Thiele and Rosenkilde, {Mette M} and IJzerman, {Adriaan P} and Heitman, {Laura H}",
note = "Copyright {\textcopyright} 2014 by The American Society for Pharmacology and Experimental Therapeutics.",
year = "2014",
doi = "10.1124/mol.114.093328",
language = "English",
volume = "86",
pages = "358--68",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "4",

}

RIS

TY - JOUR

T1 - Discovery and mapping of an intracellular antagonist binding site at the chemokine receptor CCR2

AU - Zweemer, Annelien J M

AU - Bunnik, Julia

AU - Veenhuizen, Margo

AU - Miraglia, Fabiana

AU - Lenselink, Eelke B

AU - Vilums, Maris

AU - de Vries, Henk

AU - Gibert, Arthur

AU - Thiele, Stefanie

AU - Rosenkilde, Mette M

AU - IJzerman, Adriaan P

AU - Heitman, Laura H

N1 - Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

PY - 2014

Y1 - 2014

N2 - 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.

AB - 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.

KW - Allosteric Site

KW - Amino Acid Motifs

KW - Amino Acid Sequence

KW - Animals

KW - CHO Cells

KW - COS Cells

KW - Cell Line, Tumor

KW - Cercopithecus aethiops

KW - Cricetinae

KW - Cricetulus

KW - Humans

KW - Imidazoles

KW - Ligands

KW - Molecular Sequence Data

KW - Point Mutation

KW - Protein Binding

KW - Pyrrolidines

KW - Receptors, CCR2

KW - Sulfonamides

U2 - 10.1124/mol.114.093328

DO - 10.1124/mol.114.093328

M3 - Journal article

C2 - 25024169

VL - 86

SP - 358

EP - 368

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 4

ER -

ID: 137817839