Phospholipid distribution in the microenvironment of the immunoglobulin E-receptor from rat basophilic leukemia cell membrane.
It has been previously found that lipids were required to maintain intact the tetrameric structure of the receptor for immunoglobulin E (IgE) (Fc epsilon R) in detergent solutions [Rivnay, B., Rossi, G., Henkart, M., & Metzger, H. (1984) J. Biol. Chem. 259, 1212-1217, and references cited therein]. Failure of commercially obtained lipids to provide sufficient protection, however, underscored the necessity for development of additional analytical approaches. In order to identify the phospholipid distribution in the intimate natural environment of this receptor, both the plasma membrane vesicles and the ligand-receptor complex (IgE-Fc epsilon R) have been isolated by affinity chromatography. The phospholipids of both preparations were compared. After extensive washing with detergent lipid micelles, IgE-Fc epsilon R retained 0.1-1% of the total phospholipids in the purified plasma membrane. The receptor-bound lipids were shown to contain phosphatidylcholine and sphingomyelin; the content of the latter lipid was enriched 2-5-fold compared with that in the plasma membranes. This pattern was observed with several detergents employed for purification and under a variety of experimental conditions. In light of the general distribution of choline phospholipids in the outer leaflet of plasma membranes, this enrichment may not be a characteristic of this particular receptor exclusively. These observations should be particularly helpful in studies on aggregation-induced functions of the isolated Fc epsilon receptor. In general, the methods employed enable isolation of purified and lipid-protected integral proteins and also provide an appropriate reference source of intact membrane vesicles. These qualities render this approach useful in similar studies of other membrane proteins.
D000818:Animals; D001491:Basophils / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D007073:Immunoglobulin E / Q000378:metabolism*; D007942:Leukemia, Experimental / Q000276:immunology*; D008563:Membrane Lipids / Q000032:analysis*; D051379:Mice; D008823:Micelles; D008861:Microsomes / Q000276:immunology; D010743:Phospholipids / Q000032:analysis*; D051381:Rats; D011961:Receptors, Fc / Q000378:metabolism*; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000378:metabolism*
D008563:Membrane Lipids; D008823:Micelles; D010743:Phospholipids; D011961:Receptors, Fc; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D007073:Immunoglobulin E