Characterization of a human cDNA that encodes a functional receptor for platelet activating factor.
We have cloned a cDNA for the platelet activating factor (PAF) receptor by screening an HL-60 granulocyte cDNA library with degenerate oligonucleotide probes based on conserved sequences of rhodopsin-type receptors. The 342-amino acid receptor contains 7 putative transmembrane domains, but lacks sites for N-linked glycosylation at the N-terminus. Stably transfected fibroblasts expressing the cloned cDNA responded to sub-nanomolar PAF stimulation with calcium mobilization, which could be inhibited by the PAF antagonist L-659,989. The PAF receptor message was detected as a single species of approximately 4 kb in human placenta, lung, and differentiated HL-60 granulocytes. Expression of the cloned cDNA in undifferentiated HL-60 cells devoid of endogenous PAF receptor resulted in specific and saturable binding of the PAF antagonist WEB 2086 with a dissociation constant of 30.7 nM.
Ye
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RD
Department of Immunology, Scripps Research Institute, La Jolla, CA 92037.
Prossnitz
E R
ER
Zou
A H
AH
Cochrane
C G
CG
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D005663:Furans; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D010972:Platelet Activating Factor / Q000235:genetics / Q000302:isolation & purification*; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface / Q000235:genetics / Q000302:isolation & purification*; D043562:Receptors, G-Protein-Coupled; D014162:Transfection
D005663:Furans; D015345:Oligonucleotide Probes; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor; C054893:L 659989; D004247:DNA