Identification and characterization of a rat macrophage inflammatory protein-1alpha receptor.
A macrophage inflammatory protein-1alpha (MIP-1alpha) receptor has been cloned from rat (Rattus norvegicus) genomic DNA by PCR using oligonucleotides based on the mouse CCR1 DNA sequence and expressed in HEK293 cells to enable its characterization. The receptor was cloned three times in independent experiments to generate a consensus sequence. The rat ccrl receptor amino acid sequence is 92% identical to mouse and 80% identical to human CCR1. A consensus clone was transfected into HEK293 cells using the expression vector pIRES, and stable receptor expressing cell lines were isolated. In competitive receptor binding assays using iodinated human MIP-1alpha, rat ccrl binds hMIP-1alpha, hMIP-1beta, and hMCP-3, but not hRANTES or human interleukin-8 (hIL-8). We have been unable to demonstrate calcium mobilization by rat ccrl in HEK293 cells using human chemokines as ligands. Therefore, we have adopted lowercase nomenclature for the receptor until signaling is observed. This receptor and cell line may be of use in the preclinical development of CCR1 antagonists.
Waller
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A
British Biotech Pharmaceuticals, Ltd, Oxford, UK.
Nayee
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P
Czaplewski
L G
LG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D020013:Calcium Signaling; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018925:Chemokines / Q000494:pharmacology / Q000502:physiology; D016384:Consensus Sequence; D004247:DNA / Q000097:blood; D006801:Humans; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D011917:Rats, Inbred Lew; D054389:Receptors, CCR1; D019707:Receptors, Chemokine / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014162:Transfection
C515300:CCR1 protein, human; C515301:Ccr1 protein, mouse; C515302:Ccr1 protein, rat; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D018925:Chemokines; D019402:Macrophage Inflammatory Proteins; D054389:Receptors, CCR1; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; C080633:macrophage inflammatory protein 1alpha receptor; D004247:DNA