Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Blotting, Northern
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Cell Line
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Cloning, Molecular
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DNA / analysis*
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Furans
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Humans
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Molecular Sequence Data
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Oligonucleotide Probes
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Open Reading Frames
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Platelet Activating Factor / genetics
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Platelet Activating Factor / isolation & purification*
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Platelet Membrane Glycoproteins*
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Receptors, Cell Surface / genetics
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Receptors, Cell Surface / isolation & purification*
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Receptors, G-Protein-Coupled*
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Transfection
Substances
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Furans
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Oligonucleotide Probes
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Platelet Activating Factor
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Platelet Membrane Glycoproteins
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Receptors, Cell Surface
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Receptors, G-Protein-Coupled
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platelet activating factor receptor
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L 659989
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DNA
Associated data
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GENBANK/M64053
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GENBANK/M64054
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GENBANK/M64055
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GENBANK/M64056
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GENBANK/M64057
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GENBANK/M64058
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GENBANK/M64059
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GENBANK/M64060
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GENBANK/M80436
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GENBANK/S60136