Synthesis of surface immunoglobulin E receptor in Xenopus oocytes by translation of mRNA from rat basophilic leukemia cells.
Immunoglobulin E-binding activity was expressed in Xenopus oocytes injected with mRNA from rat basophilic leukemia cells which possess abundant immunoglobulin E (IgE) receptor. Such activity was demonstrated with intact oocytes by their binding of 125I-labeled mouse monoclonal IgE. Binding activity was specific as shown by the total inhibition of 125I-IgE binding by unlabeled IgE but not by unlabeled IgG1. The relevance of the IgE-binding activity to the IgE receptor was also supported by the absence of this activity in oocytes injected with mRNA from cells lacking surface IgE receptors. mRNA coding for the IgE-binding activity was enriched in fractions sedimenting at 13.5 S in sucrose density gradients. From oocytes injected with rat basophilic leukemia mRNA, two major polypeptides were isolated by affinity purification on IgE immunoadsorbent. One (Mr = 31,000) is equivalent in size to the previously identified "receptor-associated protein;" the other (Mr = 40,000) is speculated to be a partially glycosylated or unglycosylated form of the alpha subunit of the IgE receptor. The binding of IgE-coated fluorescent microspheres by oocytes injected with rat basophilic leukemia mRNA demonstrated the surface expression of the IgE-binding proteins.
D000818:Animals; D001491:Basophils / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D005260:Female; D007073:Immunoglobulin E / Q000378:metabolism*; D007942:Leukemia, Experimental / Q000276:immunology*; D009865:Oocytes / Q000378:metabolism*; D011061:Poly A / Q000235:genetics; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000235:genetics*; D014981:Xenopus
D012333:RNA, Messenger; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D011061:Poly A; D007073:Immunoglobulin E; D012313:RNA