title: A 23 kDa membrane glycoprotein bearing NeuNAc alpha 2-3Gal beta 1-3GalNAc O-linked carbohydrate chains acts as a receptor for Streptococcus sanguis OMZ 9 on human buccal epithelial cells.
abstract: Streptococcus sanguis colonizes several human oral surfaces, including both hard and soft tissues. Large salivary mucin-like glycoproteins bearing sialic acid residues are known to bind various S.sanguis strains. However, the molecular basis for the adhesion of S.sanguis to human buccal epithelial cells (HBEC) has not been established. The present study shows that S.sanguis OMZ 9 binds to exfoliated HBEC in a sialic acid-sensitive manner. The desialylation of such cells invariably abolishes adhesion of S.sanguis OMZ 9 to the cell surface. A soluble glycopeptide bearing short sialylated O-linked carbohydrate chains behaves as a potent inhibitor of the attachment of S.sanguis OMZ 9 to exfoliated HBEC. The resialylation of desialylated HBEC with CMP-sialic acid and Gal beta 1,3GalNAc alpha 2,3-sialyltransferase specific for O-glycans restores the receptor function for S.sanguis OMZ 9, whereas a similar cell resialylation with the Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase specific for N-glycans is without effect. Finally, the same resialylation reaction carried out with CMP-9-fluoresceinyl-sialic acid as a substrate yields exfoliated HBEC bearing fluorescence on a single 23 kDa protein, when using the alpha 2,3-sialyltransferase as the catalyst. The latter finding demonstrates that this 23 kDa cell surface glycoprotein bears NeuNAc alpha 2-3Gal beta 1-3GalNAc O-linked sugar chains, a carbohydrate sequence which is recognized by S.sanguis OMZ 9 on exfoliated HBEC. In similar experiments carried out with a buccal carcinoma cell line termed SqCC/Y1, S.sanguis OMZ 9 did not attach in great numbers to such cultured cells, and these cells were shown to not express membrane glycoprotein bearing alpha 2,3-sialylated O-linked carbohydrate chains.
authors.affiliation: Nestlé Research Centre, Nestec Limited, Lausanne, Switzerland.
authors.forename: J R
authors.initials: JR
authors.lastname: Neeser
authors.forename: R C
authors.initials: RC
authors.lastname: Grafström
authors.forename: A
authors.initials: A
authors.lastname: Woltz
authors.forename: D
authors.initials: D
authors.lastname: Brassart
authors.forename: V
authors.initials: V
authors.lastname: Fryder
authors.forename: B
authors.initials: B
authors.lastname: Guggenheim
mesh_terms: D001422:Bacterial Adhesion; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000378:metabolism / Q000502:physiology; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009061:Mouth Mucosa / Q000382:microbiology* / Q000502:physiology; D009062:Mouth Neoplasms; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D012799:Sialyltransferases; D013298:Streptococcus sanguis / Q000502:physiology* / Q000648:ultrastructure; D014407:Tumor Cells, Cultured; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
chemical_list: D006020:Glycopeptides; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D012799:Sialyltransferases; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase