Purification and characterization of the specific binding protein for platelet activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) from human platelets.
The binding protein to platelet activating factor (PAF) was solubilized with 2% Triton X-100 from human platelet membranes and purified approximately 23-fold by sequential chromatography on DEAE-cellulose, CM-cellulose and Sephadex G-200. The final preparation migrated as a single protein band corresponding to a radioactive peak of [3H]PAF bound on polyacrylamide disc gel electrophoresis. The molecular weight of the native form of the binding protein was estimated to be approximately 160,000 daltons by Sephadex G-200 column chromatography, and its isoelectric point was near 8.0. Exposure of the platelet membranes to an excess of unlabeled PAF diminished the binding of [3H]PAF.
D001792:Blood Platelets / Q000032:analysis*; D002352:Carrier Proteins / Q000302:isolation & purification*; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006801:Humans; D007526:Isoelectric Point; D008970:Molecular Weight; D010972:Platelet Activating Factor / Q000378:metabolism*; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D043562:Receptors, G-Protein-Coupled; D012995:Solubility
D002352:Carrier Proteins; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor