Structural studies of Fc receptors. IV. Structure required for phospholipids for reconstitution of the delipidated Fc receptor of macrophages.
To analyze the interaction of the macrophage Fc receptor with phospholipids, we established an experimental system for delipidation of Fc receptor fraction and reconstitution of the Fc receptor activity in phospholipid vesicles. The separation of FcR from membrane phospholipids was achieved by ion exchange chromatography on DEAE-cellulose of the anionic detergent-lysate of the crude membrane fraction of guinea pig macrophages in the presence of detergent. The separation was based on the difference in charge between the complex of FcR and the anionic detergent and that of phospholipids and the detergent. The FcR fraction free of phospholipids showed no FcR activity as assessed in terms of its ability to inhibit the binding of labeled soluble immune complex of IgG2 antibody to macrophages, but the same fraction showed a definite activity when associated with phospholipids. This fraction was shown to contain a component of 44,000 daltons that is susceptible to surface-labeling and binds to IgG2-Sepharose in the affinity chromatography, indicating this component to be the Fc receptor. Reconstitution experiments with this fraction showed that phosphatidylcholine is the most effective phospholipid to reconstitute the FcR activity among those tested. Phosphatidylserine, phosphatidylinositol, and sphingomyelin were ineffective, while phosphatidylethanolamine showed a moderate effect. The inactivating effect of phospholipase C treatment on the Fc receptor activity of the membrane was shown to be due to the cleavage of phospholipids in the membrane but not due to modification of the Fc receptor molecule itself.
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000276:immunology; D006168:Guinea Pigs; D008264:Macrophages / Q000276:immunology*; D008563:Membrane Lipids / Q000502:physiology*; D008970:Molecular Weight; D010743:Phospholipids / Q000502:physiology*; D011961:Receptors, Fc / Q000302:isolation & purification / Q000378:metabolism*; D010738:Type C Phospholipases / Q000494:pharmacology
D008563:Membrane Lipids; D010743:Phospholipids; D011961:Receptors, Fc; D010738:Type C Phospholipases