Glycosylation of the tandem repeat unit of the MUC2 polypeptide leading to the synthesis of the Tn antigen.
A synthetic peptide corresponding to the human MUC2 tandem repeat domain containing 14 Thr residues was glycosylated in vitro using UDP-GalNAc and microsomal membranes of the colorectal cancer cell line, LS180. The products were fractionated by reverse phase HPLC, which gave seven glycopeptide fractions. Their molecular weights were estimated by matrix-assisted laser desorption/ionization mass spectrometry, the values obtained corresponding to glycopeptides containing from one to ten GalNAc residues. On solid phase radioimmunoassaying involving a monoclonal anti-Tn antibody (MLS128), it was found that the glycopeptides containing nine or ten GalNAc residues were strongly immunoreactive, whereas the glycopeptides containing less than six GalNAc residues were inactive, indicating that a cluster of GalNAc-Thr is essential for the Tn antigenicity.
Inoue
M
M
Department of Biochemistry, Faculty of Engineering, Kyoto Sangyo University, Kita-ku, Kyoto, 603, Japan.
Yamashina
I
I
Nakada
H
H
D000116:Acetylgalactosamine / Q000032:analysis; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000096:biosynthesis* / Q000276:immunology; D015179:Colorectal Neoplasms / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D055262:Mucin-2; D009077:Mucins / Q000737:chemistry*; D017350:N-Acetylgalactosaminyltransferases; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism; D010455:Peptides; D011863:Radioimmunoassay / Q000379:methods; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014407:Tumor Cells, Cultured; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000378:metabolism; D000097763:Polypeptide N-acetylgalactosaminyltransferase
D000116:Acetylgalactosamine; D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D006020:Glycopeptides; D055262:Mucin-2; D009077:Mucins; D017350:N-Acetylgalactosaminyltransferases; D010446:Peptide Fragments; D010455:Peptides; D014536:Uridine Diphosphate N-Acetylgalactosamine; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C524625:MUC2 protein, human; C040286:Tn antigen