Glycosylation of a novel member of the immunoglobulin gene superfamily expressed in rat carcinoma cell lines.
MAb E4 recognizes a 66-kDa glycoprotein, pE4, which is a member of the immunoglobulin gene superfamily. This protein is expressed at the cell surface in rat colon and mammary carcinomas, but only in trace amounts in normal adult rat tissues. Since expression of aberrant carbohydrate structures is often associated with malignant transformation, glycosylation of pE4 was analyzed. Reactivity of lectins with pE4 suggested the absence of N-acetylneuraminic acid, terminal galactose and O-linked glycan, and the presence of N-linked glycans. Tunicamycin treatment reduced the binding of MAb E4 to cancer cells suggesting that the E4 epitope is at least partially glycosylated. Digestions with neuraminidases, O-glycosidase and peptide-N-glycosidase F confirmed these results. Pronase treatment abolished the binding of MAb E4, indicating that E4 epitope involves not only a carbohydrate determinant but also a peptide moiety. Mild periodate oxidation abolished the binding of MAb E4, indicating that non-reducing terminus carbohydrates are part of the E4 epitope. Neutral sugar analysis revealed the absence of galactose and the presence of fucose. Since fucose is sensitive to periodate oxidation, this sugar could be the carbohydrate part of the determinant recognized by MAb E4. Reactivity of lectins specific for fucose indicated the presence of alpha(1-6)-fucose on pE4.
Chadéneau
C
C
INSERM CJF90-11, Institut de Biologie, Nantes, France.
Le Moullac
B
B
Cornu
G
G
Meflah
K
K
Denis
M G
MG
D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism*; D002240:Carbohydrate Sequence; D003110:Colonic Neoplasms / Q000235:genetics* / Q000378:metabolism*; D010450:Endopeptidases / Q000494:pharmacology; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D005643:Fucose / Q000378:metabolism; D005803:Genes, Immunoglobulin; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006031:Glycosylation; D037102:Lectins / Q000378:metabolism; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000494:pharmacology; D011134:Polysaccharides / Q000032:analysis; D051381:Rats; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; D006023:Glycoproteins; D037102:Lectins; D008565:Membrane Proteins; D011134:Polysaccharides; C092931:Pvr protein, rat; D010504:Periodic Acid; D014415:Tunicamycin; D005643:Fucose; D006026:Glycoside Hydrolases; D010450:Endopeptidases