RAT BASOPHILIC LEUKEMIA CELL RECEPTOR FOR IGE .1. CHARACTERIZATION AS A GLYCOPROTEIN

Abstract

Rat basophilic leukemia cells were labeled enzymically with 125I or biosynthetically by culture in the presence of 14C-glucosamine or 3H-amino acids and then extracted with NP-40 [Nonidet P-40]. IgE[immunoglobulin E]-anti-IgE precipitates insolubilized a radiolabeled macromolecule from these extracts largely or entirely absent in control IgG-anti-IgG precipitates. When specific precipitates were boiled in sodium dodecyl sulfate (SDS) and analyzed by polyacrylamide gel electrophoresis in the presence of SDS, most of the 14C or 125I radioactivity was in the area corresponding to an apparent MW of 60,000-70,000 in 5.9% gels. In 10% and 12% gels, faster mobility was demonstrated indicating an atypical electrophoretic behavior often associated with glycoproteins and a presumptive MW of 50,000 or less. Since only IgE-containing precipitates localized label in this region and since such precipitates from cells saturated with IgE prior to surface iodination failed to show this band, the labeled macromolecule appears to be the IgE receptor itself. Analysis of the acid hydrolysates of precipitated 14C radioactivity demonstrated that label was entirely in hexosamines and sialic acid. 125I and 14C labels in the receptor region were eliminated almost completely with pepsin and pronase and to a lesser extent with trypsin.