Deciphering the Non-Equivalence of Serine and Threonine O-Glycosylation Points: Implications for Molecular Recognition of the Tn Antigen by an anti-MUC1 Antibody

Angew Chem Int Ed Engl. 2015 Aug 17;54(34):9830-4. doi: 10.1002/anie.201502813. Epub 2015 Jun 26.

Abstract

The structural features of MUC1-like glycopeptides bearing the Tn antigen (α-O-GalNAc-Ser/Thr) in complex with an anti MUC-1 antibody are reported at atomic resolution. For the α-O-GalNAc-Ser derivative, the glycosidic linkage adopts a high-energy conformation, barely populated in the free state. This unusual structure (also observed in an α-S-GalNAc-Cys mimic) is stabilized by hydrogen bonds between the peptidic fragment and the sugar. The selection of a particular peptide structure by the antibody is thus propagated to the carbohydrate through carbohydrate/peptide contacts, which force a change in the orientation of the sugar moiety. This seems to be unfeasible in the α-O-GalNAc-Thr glycopeptide owing to the more limited flexibility of the side chain imposed by the methyl group. Our data demonstrate the non-equivalence of Ser and Thr O-glycosylation points in molecular recognition processes. These features provide insight into the occurrence in nature of the APDTRP epitope for anti-MUC1 antibodies.

Keywords: X-ray diffraction; antibodies; conformation analysis; glycopeptides; molecular recognition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology*
  • Antigens, Tumor-Associated, Carbohydrate / chemistry
  • Antigens, Tumor-Associated, Carbohydrate / immunology*
  • Glycosylation
  • Models, Molecular
  • Molecular Conformation
  • Mucin-1 / chemistry
  • Mucin-1 / immunology*
  • Serine / chemistry
  • Serine / immunology*
  • Serine / metabolism
  • Threonine / chemistry
  • Threonine / immunology*
  • Threonine / metabolism

Substances

  • Antibodies, Monoclonal
  • Antigens, Tumor-Associated, Carbohydrate
  • Mucin-1
  • Tn antigen
  • Threonine
  • Serine