Monoclonal antibodies against defined epitopes of the human transferrin receptor cytoplasmic tail.
Three murine monoclonal antibodies (mAbs) against the 61-residue amino-terminal cytoplasmic tail of the human transferrin receptor (TR) have been produced by immunization of mice with recombinant human TR produced in a baculovirus expression system. Mutant human TRs expressed in chick embryo fibroblasts (CEFs) with point mutations or deletions in their cytoplasmic tails have been used to map the epitopes defined by each of the mAbs. One mAb, H68.4, previously shown to block receptor internalization, binds proximal to the carboxy-terminal side of the YTRF internalization signal of TR. The second mAb, H73.2, binds near to the carboxy-terminal side of the H68.4 epitope, whereas the third mAb, 160.1, binds closer to the transmembrane region. H68.4 and H73.2 are auto-antibodies consistent with their epitopes mapping to a region of the human TR that has an identical amino acid sequence to the mouse TR. All three mAbs crossreact with the cytoplasmic tail of Chinese hamster TR. Double labelling of recombinant human TRs on chick embryo fibroblast (CEF) cell membrane preparations with B3/35 and H68.4 antibody-gold conjugates established that receptors in clathrin-coated pits were not labeled with H68.4, implying that associated coated pit proteins may block binding of this mAb.
White
S
S
Department of Cancer Biology, Salk Institute for Biological Studies, San Diego, CA 92186-5800.
Miller
K
K
Hopkins
C
C
Trowbridge
I S
IS
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002642:Chick Embryo; D003034:Coated Pits, Cell-Membrane / Q000276:immunology; D003429:Cross Reactions; D003593:Cytoplasm / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011990:Receptors, Transferrin / Q000276:immunology*; D016415:Sequence Alignment; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011990:Receptors, Transferrin