Studies of the IgE binding sites to rat mast cell receptor with proteolytic fragments and with a monoclonal antibody directed against epsilon heavy chain: evidence that the combining sites are located in the C epsilon 3 domain.
The binding sites of rat IgE to mast cell receptor were investigated by the use of proteolytic fragments and a monoclonal antibody to epsilon chain (MARE-1). Three main fragments were characterized by short-time papain digestion of IgE: F(ab')2-E, a fragment related to the C, 4 domain, and an asymmetric fragment corresponding probably to an IgE molecule with one proteolyzed C, 3 domain. Neither F(ab')2-E nor C, 4 could interfere with the binding of IgE to rat mast cells. These two fragments did not show significant polymerization upon heating at 56 degrees C, while large amounts of polymers were produced from whole IgE, MARE-1 monoclonal antibody was found to react neither with F(ab')2 nor with C, 4, thereby suggesting its interaction with the C, 3 domain. MARE-1 was found to inhibit partially (about 55%) the binding of IgE to its receptor. Taken together the results indicate that the binding sites of IgE to rat mast cell receptor are located within the C, 3 domain. In addition, isolation of the C, 4 domain will be useful to evaluate its participation in the affinity of IgE to receptors of other cells such as lymphocytes or macrophages.
D000818:Animals; D001666:Binding Sites, Antibody / Q000235:genetics*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005802:Genes, MHC Class II; D006358:Hot Temperature; D007128:Immunoglobulin Fragments / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007139:Immunoglobulin epsilon-Chains / Q000235:genetics*; D008407:Mast Cells / Q000276:immunology*; D008970:Molecular Weight; D010206:Papain / Q000494:pharmacology; D051381:Rats; D011961:Receptors, Fc / Q000235:genetics*; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000235:genetics*
D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007139:Immunoglobulin epsilon-Chains; D011961:Receptors, Fc; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D010206:Papain