Monoclonal antibodies to either domain of ovotransferrin block binding to transferrin receptors on chick reticulocytes.
Monoclonal antibodies produced to both chicken ovotransferrin and to the isolated N- and C-terminal half-molecule domains of ovotransferrin have been used to probe the interaction of ovotransferrin with its specific receptor on chick embryo red blood cells. Two antibodies to epitopes on the N-terminal domain and one antibody to an epitope on the C-terminal domain were able to block the binding of 125I-labeled diferric ovotransferrin to the receptor. When the cellular surface receptors were first saturated with ovotransferrin at 0 degrees C, none of these antibodies bound to the cell-associated ovotransferrin. This suggests that the antibodies are to epitopes which lie very near to, or in the regions of, the two domains which interact with receptor. The same three antibodies also blocked the binding to the receptor of ovotransferrin associated in situ from the isolated N- and C-terminal half-molecule domains. A fourth antibody did not block binding to receptor of 125I-labeled diferric ovotransferrin or the associated domains; furthermore, it was able to bind to ovotransferrin bound to the cell surface at 0 degrees C. This antibody thus appears to recognize an epitope remote from the receptor binding region of ovotransferrin. Additional evidence for the requirement of the presence of both domains of ovotransferrin to effect binding to the transferrin receptor on chick reticulocytes was obtained with a fifth antibody which recognized only the N-terminal half-molecule domain but not holo-ovotransferrin. Although this antibody had no effect on the binding of 125I-labeled ovotransferrin to cells, it blocked binding to receptor of the associated domains of ovotransferrin, presumably by inhibiting the association of the two domains.
D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002642:Chick Embryo; D003207:Conalbumin / Q000097:blood / Q000276:immunology*; D004527:Egg Proteins / Q000276:immunology*; D000939:Epitopes / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D011990:Receptors, Transferrin / Q000378:metabolism*; D012156:Reticulocytes / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004527:Egg Proteins; D000939:Epitopes; D046911:Macromolecular Substances; D011990:Receptors, Transferrin; D003207:Conalbumin