Glycoconjugate Journal

, Volume 13, Issue 5, pp 717–726 | Cite as

Benzyl-N-acetyl-α-d-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation

  • Philippe Delannoy
  • Isabelle Kim
  • Nathalie Emery
  • Carmen de Bolos
  • Andre Verbert
  • Pierre Degand
  • Guillemette Huet
Papers

Abstract

We have analysed the mucins synthesized by the HT-29 MTX cell subpopulation, derived from the HT-29 human colon carcinoma cells through a selective pressure with methotrexate (Lesuffleuret al., 1990,Cancer Res50: 6334–43), in the presence of benzyl-N-acetyl-α-galactosaminide (GalNAcα-O-benzyl), which is a potential competitive inhibitor of the β1,3-galactosyltransferase that synthesizes the T-antigen. The main observation was a 13-fold decrease in the sialic acid content of mucins after 24 h of exposure to 5mm GalNAcα-O-benzyl. This effect was accompanied by an increased reactivity of these mucins to peanut lectin, testifying to the higher amount of T-antigen. The second observation was a decrease in the secretion of the mucins by GalNAcα-O-benzyl treated cells. The decrease in mucin sialyation was achieved through thein situ β-galactosylation of GalNAcα-O-benzyl into Galβ1–3GalNAcα-O-benzyl, which acts as a competitive substrate of Galβ1–3GalNAc α2,3-sialyltransferase, as shown by the intracellular accumulation of NeuAcα2–3Galβ1–3GalNAcα-O-benzyl in treated cells.

Keywords

HT-29 cells mucins aryl-glycosides O-glycosylation sialyltransferases 

Abbreviations

BSM

bovine submaxillary mucin

MTX

methotrexate

PBS

sodium phosphate 10mm, NaCl 0.15m, pH 7.4 buffer

pNp

p-nitrophenol

TBS

Tris/HCl 10mm, NaCl 0.15m, pH 7.4 buffer

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Copyright information

© Chapman & Hall 1996

Authors and Affiliations

  • Philippe Delannoy
    • 1
  • Isabelle Kim
    • 2
  • Nathalie Emery
    • 2
  • Carmen de Bolos
    • 3
  • Andre Verbert
    • 1
  • Pierre Degand
    • 2
  • Guillemette Huet
    • 2
  1. 1.Laboratoire de Chimie Biologique, Unité Mixte de Recherche du CNRS no 111Université des Sciences et Technologies de LilleVilleneuve d'AscqFrance
  2. 2.INSERM U-377LilleFrance
  3. 3.Institut Municipal d'Investigacio MedicaUniversitat Autonoma de BarcelonaSpain

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