Structural studies of fc receptors. II. The effect of dissociation and reassociation of phospholipids on the activity of Fc gamma receptors of macrophages.
The receptor for the Fc portion of immunoglobulin G (Fc receptor) of guinea pig macrophages was solubilized with a detergent and partially delipidated to the point where the ligand binding activity was essentially lost. Delipidation of the Fc receptor was done by fractionating the macrophage lysate by gel filtration in the presence of detergent. The elution behavior of Fc receptor-detergent complex and phospholipid-detergent mixed micelles varied depending on the kind of detergents used for membrane solubilization and for gel filtration. Separation of phospholipids from Fc receptor was best achieved when octylglucoside-solubilized fraction was chromatographed on Sepharose CL-6B in the presence of deoxycholate; the phospholipid peak emerged at Kav = 0.55 and the Fc receptor at Kav = 0.45. The fraction of Kav = 0.45 showed only a marginal activity when the activity was measured after removal of detergents, but activity was clearly shown when phospholipid fraction was added to this fraction prior to removal of the detergents. Reappearance of the Fc receptor activity was shown to be due to association of phospholipids with the Fc receptor. Three kinds of phospholipids with different polar head groups examined, phosphatidylcholine, phosphatidylserine, and phosphatidylethanolamine, were all able to reconstitute active Fc receptor, although phosphatidylethanolamine was somewhat less effective than the others. Thus, our study demonstrated the amphipathic nature of the Fc receptor, the binding of which is dependent on the interaction with phospholipids.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D003902:Detergents; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008264:Macrophages / Q000378:metabolism*; D010743:Phospholipids / Q000494:pharmacology*; D011961:Receptors, Fc / Q000378:metabolism*; D017452:Receptors, IgG; D011971:Receptors, Immunologic / Q000378:metabolism*; D012995:Solubility
D003902:Detergents; D010743:Phospholipids; D011961:Receptors, Fc; D017452:Receptors, IgG; D011971:Receptors, Immunologic