The N-terminal extracellular segments of the chemokine receptors CCR1 and CCR3 are determinants for MIP-1alpha and eotaxin binding, respectively, but a second domain is essential for efficient receptor activation.
CCR1 and CCR3 are seven-transmembrane domain G protein-coupled receptors specific for members of the CC chemokine subgroup of leukocyte chemoattractants. Both have been implicated in the inflammatory response, and CCR3, through its expression on eosinophils, basophils, and Th2 lymphocytes, may be especially important in allergic inflammation. CCR1 and CCR3 are 54% identical in amino acid sequence and share some ligands but not others. In particular, macrophage inflammatory protein 1alpha (MIP-1alpha) is a ligand for CCR1 but not CCR3, and eotaxin is a ligand for CCR3 but not CCR1. To map ligand selectivity determinants and to guide rational antagonist design, we analyzed CCR1:CCR3 chimeric receptors. When expressed in mouse pre-B cells, chimeras in which the N-terminal extracellular segments were switched were both able to bind both MIP-1alpha and eotaxin, but in each case, binding occurred via separate sites. Nevertheless, neither MIP-1alpha nor eotaxin were effective agonists at either chimeric receptor in either calcium flux or chemotaxis assays. These data are consistent with a multi-site model for chemokine-chemokine receptor interaction in which one or more subsites determine chemokine selectivity, but others are needed for receptor activation. Agents that bind to the N-terminal segments of CCR1 and CCR3 may be useful in blocking receptor function.
Krebs Institute, University of Sheffield, Western Bank, Sheffield S10 2TN, United Kingdom. j.pease@ic.ac.uk
J E
JE
Pease
J
J
Wang
P D
PD
Ponath
P M
PM
Murphy
D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology; D001665:Binding Sites / Q000502:physiology; D054413:Chemokine CCL11; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D019742:Chemokines, CC / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding / Q000502:physiology; D054389:Receptors, CCR1; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology; D014407:Tumor Cells, Cultured
C515457:Ccl11 protein, mouse; C515301:Ccr1 protein, mouse; C515351:Ccr3 protein, mouse; D054413:Chemokine CCL11; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D019742:Chemokines, CC; D016207:Cytokines; D019402:Macrophage Inflammatory Proteins; D054389:Receptors, CCR1; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins; D019204:GTP-Binding Proteins