Antigen-induced cross-linking of the IgE receptor leads to an association with the detergent-insoluble membrane skeleton of rat basophilic leukemia (RBL-2H3) cells.
Cross-linking of the IgE receptor on the surface of rat basophilic leukemia cells by multivalent Ag (DNP-BSA) causes a rapid conversion to a detergent-insoluble form. There is a concurrent increase in the amount of filamentous actin associated with the plasma membrane. Both the degree of receptor detergent insolubility and the rise in F-actin content are rapid with a half-maximal response of less than 1 min and can be rapidly reversed by the addition of monovalent Ag (DNP-lysine). These two early steps in the triggering of rat basophilic leukemia cells can be dissociated from each other by pretreatment of the cells with either cytochalasin or sodium azide. These reagents block the increase in F-actin but have no effect on receptor detergent insolubility. This indicates that microfilaments are not responsible for detergent insolubility of the receptor and that it may be the membrane skeleton that is interacting with the complex. This was further confirmed by the finding that cross-linking of the IgE receptors on the surface of purified plasma membranes also leads to detergent insolubility of the receptor. Therefore, all of the components necessary for detergent insolubility of the receptor are present in the plasma membrane, and cytoplasmic components are not needed. These results suggest that detergent insolubility and immobility of the cross-linked receptors are caused by multivalent interaction with the membrane skeleton. Actin filaments may then interact with these receptor-membrane skeletal complexes in order to produce large scale clustering and capping. The membrane skeleton may therefore be acting as an intermediate structure between the cell-surface receptors and microfilaments.
Apgar
J R
JR
Division of Membrane Biology, Medical Biology Institute, La Jolla, CA 92037.
D008841:Actin Cytoskeleton / Q000502:physiology; D000199:Actins / Q000378:metabolism; D000818:Animals; D000941:Antigens / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000737:chemistry*; D003599:Cytoskeleton / Q000502:physiology*; D003902:Detergents / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D051381:Rats; D011961:Receptors, Fc / Q000737:chemistry*; D017455:Receptors, IgE; D012701:Serotonin / Q000378:metabolism; D012995:Solubility; D014407:Tumor Cells, Cultured
D000199:Actins; D000941:Antigens; D000944:Antigens, Differentiation, B-Lymphocyte; D003902:Detergents; D008565:Membrane Proteins; D011961:Receptors, Fc; D017455:Receptors, IgE; D012701:Serotonin