N- and O-linked oligosaccharides completely lack galactose residues in the gms1och1 mutant of Schizosaccharomyces pombe

Appl Microbiol Biotechnol. 2010 Mar;86(1):263-72. doi: 10.1007/s00253-009-2297-9. Epub 2009 Oct 21.

Abstract

Unlike their counterparts in budding yeast Saccharomyces cerevisiae, the glycoproteins of Schizosaccharomyces pombe contain, in addition to alpha-D-mannose (Man), a large number of alpha-D-galactose (Gal) residues. In both yeasts, large outer chains are attached to the oligosaccharide cores of glycoproteins during export via Golgi. Formation of the yeast-specific large outer chain is initiated by alpha-1,6-mannosylatransferase encoded by the och1+ gene, the disruption of which blocked outer chain elongation. We previously reported that N-linked oligosaccharide structures of S. pombe och1Delta mutant consisted of Gal(2-6)Man(9)GlcNAc(2) with alpha-linked Gal residues attached to the core oligosaccharide moiety. The disruption of gms1+, a gene encoding the UDP-galactose transporter required for the synthesis of galactomannan, abolished cell surface galactosylation in S. pombe. In this study, we constructed a gms1Deltaoch1Delta double mutant and determined the N- and O-linked oligosaccharide structures present on the cell surface. Oligosaccharides were liberated from glycoproteins by hydrazinolysis and labeled with the fluorophore, 2-aminopyridine. The pyridylaminated N-linked oligosaccharides were analyzed by high-performance liquid chromatography in combination with alpha1,2-mannosidase digestion and partial acetolysis. These analyses revealed that the N-linked oligosaccharides of gms1Deltaoch1Delta cells consisted of alpha1,2-linked Man-extended core oligosaccharides (Man(8-12)GlcNAc2) from which the fission yeast-specific alpha-linked Gal residues were completely absent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbohydrate Sequence
  • Galactose / analysis*
  • Galactose / chemistry
  • Mannosyltransferases / genetics*
  • Molecular Sequence Data
  • Monosaccharide Transport Proteins / genetics*
  • Mutation*
  • Oligosaccharides / chemistry*
  • Oligosaccharides / metabolism
  • Schizosaccharomyces / chemistry*
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / genetics*
  • UDP-Galactose Translocators

Substances

  • Galactose
  • Mannosyltransferases
  • Monosaccharide Transport Proteins
  • Oligosaccharides
  • Schizosaccharomyces pombe Proteins
  • UDP-Galactose Translocators
  • Och1 protein, S pombe