The effect of glycosylation on the antibody recognition of a MUC2 mucin epitope

Katalin Uray, Mamoru Mizuno, Toshiyuki Inazu, Kohtaro Goto, Ferenc Hudecz

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

MUC2 glycoprotein, produced by the epithelium of the colon and built up mainly of repeat units of 1PTTTPITTTTTVTPTPTPTGTQT23, can be overexpressed or underglycosylated in gastrointestinal diseases, e.g. in case of colon carcinoma. We have been studying the epitope structure of the MUC2 by focusing on the repeat unit with the mucin peptide specific MAb 996 monoclonal antibody. This antibody recognizes the 18PTGTQ22 sequence as minimal, and 16PTPTGTQ22 as optimal epitope within the underglycosylated glycoprotein. In this article, we aim to clarify the effect of glycosylation of the epitope on MAb 996 antibody binding including its correlation with the secondary structure of the modified peptides: glycosylation in the epitope core and in the flank. For this we have prepared the 16PTPTGTQ22 peptide glycosylated with N-acetylgalactoseamine (Tn antigen) in position 17, 19, 21, or on all three threonines. The MAb 996 antibody binding properties of the peptides were characterized in competitive ELISA experiments, and their solution secondary structure was studied by circular dichroism spectroscopy in water and in the ordered structure promoting trifluoroethanol. Our results show that glycosylation in position 19 (peptide 16PTPT(GalNAcα)GTQ22) resulted in enhanced antibody recognition and significantly altered secondary structure, while glycosylation in position 21 completely demolished the binding. These findings could be useful in determining the nature of antigen-antibody interaction, and perhaps designing synthetic peptide vaccines for tumor therapy.

Original languageEnglish
Pages (from-to)390-395
Number of pages6
JournalBiopolymers - Peptide Science Section
Volume102
Issue number5
DOIs
Publication statusPublished - Sep 1 2014

Keywords

  • Antibody binding of epitope
  • Glycosylated peptides
  • MUC2 mucin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Biomaterials
  • Organic Chemistry

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