Identification of the immunoglobulin G receptor of human platelets.
The binding site of IgG on human platelets was studied by the use of the cleavable heterobifunctional cross-linking agent N-succinimidyl (4-azidophenyldithio)propionate. Binding characteristics of the derivatized IgG were similar to normal IgG. Periodate-borohydride treatment of platelets also did not significantly alter their ability to bind IgG. N-Succinimidyl (4-azidophenyldithio)propionate was bound to IgG via a succinimidyl ester and then photolyzed in the presence of intact platelets. Their membrane glycoproteins were first tritiated by the periodate-borohydride method. The cross-linked product was analyzed by two dimensional sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis. The non-reduced first-dimension gels were subjected to 5% 2-mercaptoethanol prior to separation in the second dimension. Such gels were then evaluated by fluorography, silver staining, and counting the radioactivity of sequential gel strips in the area of cross-linking. The protein complexes at the interface between stacking and running gel were further resolved in isoelectric focusing gels. One IgG-containing band could be identified. After reduction, the constituent proteins of the cross-linked complex were analyzed by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis and subsequent immunoblotting with an antiserum against platelet membrane glycoproteins. All of these studies gave evidence of glycoprotein IIIa as the receptor of IgG. Based on the results of the different experimental approaches, we conclude that glycoprotein IIIa is the IgG receptor in human platelets.
D001386:Azides; D001792:Blood Platelets / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007457:Iodine Radioisotopes; D008565:Membrane Proteins / Q000032:analysis; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D010980:Platelet Membrane Glycoproteins; D017452:Receptors, IgG; D011971:Receptors, Immunologic / Q000032:analysis* / Q000276:immunology; D012709:Serum Albumin / Q000378:metabolism; D012794:Sialic Acids / Q000032:analysis; D014316:Tritium
D001386:Azides; D006023:Glycoproteins; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010980:Platelet Membrane Glycoproteins; D017452:Receptors, IgG; D011971:Receptors, Immunologic; D012709:Serum Albumin; D012794:Sialic Acids; D014316:Tritium; C033169:3-((4-azidophenyl)dithio)propionic N-hydroxysuccinimide; D019158:N-Acetylneuraminic Acid