A cell surface glycoprotein of rat basophilic leukemia cells close to the high affinity IgE receptor (Fc epsilon RI). Similarity to human melanoma differentiation antigen ME491.
A monoclonal antibody (mAb), AD1, was isolated that recognized a cell surface protein on rat basophilic leukemia cells (RBL-2H3). At high concentration, this antibody inhibited IgE-mediated but not calcium ionophore-induced histamine release (49% inhibition at 100 micrograms/ml). The mAb AD1 did not inhibit the binding of IgE or of several antibodies directed to the high affinity IgE receptor (Fc epsilon RI). Likewise, IgE did not inhibit mAb AD1 binding. However, several anti-Fc epsilon RI antibodies did inhibit mAb AD1 binding as intact molecules but not as Fab fragments. Therefore, the sites on the cell surface to which mAb AD1 binds are close to Fc epsilon RI. The mAb AD1 immunoprecipitated a broad, 50-60-kDa band from 125I-surface-labeled RBL-2H3 cells that upon peptide N-glycosidase F treatment was transformed into a sharp 27-kDa band. A similar 27-kDa protein was immunoprecipitated from surface-radiolabeled cells after culture with tunicamycin. Thus, the protein recognized by mAb AD1 is highly glycosylated with predominantly N-linked oligosaccharides. The N-terminal sequence of 43 amino acids was found to be different from any subunit of Fc epsilon RI but nearly identical to that of the human melanoma-associated antigen ME491. Therefore, mAb AD1 binds to a surface glycoprotein on RBL-2H3 cells sterically close to the Fc epsilon RI but distinct from the recognized subunits of the receptor.
Kitani
S
S
Clinical Immunology Section, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892.
Berenstein
E
E
Mergenhagen
S
S
Tempst
P
P
Siraganian
R P
RP
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000235:genetics; D000944:Antigens, Differentiation, B-Lymphocyte / Q000502:physiology*; D000951:Antigens, Neoplasm / Q000235:genetics; D015153:Blotting, Western; D006636:Histamine Release; D008407:Mast Cells / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000276:immunology / Q000502:physiology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins / Q000235:genetics; D011233:Precipitin Tests; D051381:Rats; D011961:Receptors, Fc / Q000502:physiology*; D017455:Receptors, IgE; D015398:Signal Transduction; D060149:Tetraspanin 30; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; C554586:CD63 protein, human; C554588:Cd63 protein, rat; D008562:Membrane Glycoproteins; D010980:Platelet Membrane Glycoproteins; D011961:Receptors, Fc; D017455:Receptors, IgE; D060149:Tetraspanin 30; D014415:Tunicamycin