The interleukin-8 receptor is encoded by a neutrophil-specific cDNA clone, F3R.
Interleukin-8 (IL-8) is one of the most potent chemotactic agents for neutrophils and has been implicated as a major mediator of inflammation. The IL-8 receptor is expressed exclusively in neutrophils and belongs to the family of G-protein-coupled receptors. In a recent paper we reported the characterization of a cDNA clone, F3R, isolated from a neutrophil cDNA library and showed that it encodes a G-protein-coupled receptor which is exclusively expressed in neutrophils. We also suggested, based on expression studies in Xenopus oocytes, that the F3R protein product is an isoform of the (fMLP) receptor (Thomas, K. M., Pyun, H. Y., and Navarro, J. (1990) J. Biol. Chem. 265, 20061-20064). In this work, the F3R receptor cDNA is expressed in monkey kidney cells (COS-7) and is shown to encode the IL-8 receptor. F3R cDNA does not encode for a fMLP receptor isoform. We show conclusively that the F3R-transfected COS-7 cells express the IL-8 receptor at a density equivalent to that observed in neutrophils. The pharmacological profile of the F3R-transfected cells is the same as that of neutrophils. The apparent Kd values for binding of 125I-IL-8 to neutrophils and F3R-transfected COS-7 cell membranes were 1.2 and 1.4 nM, respectively. Antipeptide antibodies against a partial sequence of the F3R protein product specifically immunoprecipitate the IL-8 receptor from transfected cells as well as neutrophils. The molecular characterization of the IL-8 receptor should provide the basis for further studies on the identification of the binding domain of this inflammatory receptor.
Thomas
K M
KM
Department of Physiology, Boston University School of Medicine, Massachusetts 02118.
Taylor
L
L
Navarro
J
J
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D004247:DNA; D019204:GTP-Binding Proteins / Q000378:metabolism; D000882:Haplorhini; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism*; D011233:Precipitin Tests; D012313:RNA / Q000032:analysis; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000235:genetics*; D023062:Receptors, Interleukin-8A; D014158:Transcription, Genetic; D014162:Transfection
D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D023062:Receptors, Interleukin-8A; D009240:N-Formylmethionine Leucyl-Phenylalanine; D012313:RNA; D004247:DNA; D019204:GTP-Binding Proteins