The och1 mutant of schizosaccharomyces pombe produces galactosylated core structures of n-linked oligosaccharides

Takao Ohashi, Yuka Ikeda, Naotaka Tanaka, Shin Ichi Nakakita, Shunji Natsuka, Yuko Giga-Hama, Kaoru Takegawa

Research output: Contribution to journalArticle

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Abstract

Unlike the budding yeast Saccharomyces cerevisiae, the fission yeast Schizosaccharomyces pombe synthesizes large outer chains on the N-linked oligosaccharides that consist mainly of D-Gal and D-Man residues. The fission yeast och1+ gene product has γ1,6-mannosyltransferase activity, and Och1p is the key enzyme in the initiation of outer chain elongation. Although the in vitro substrate specificity of S. pombe Och1p has been reported (Yoko-o et al., FEBS Lett., 489, 75-80 (2001)), the structure of the N-linked oligosaccharides of och1δ cells has not been investigated. In this study, we report a structural analysis of S. pombe N-linked oligosaccharides. Lectin blot analysis indicated that galactose residues were attached to the cell surface glycoproteins of the och1δ cells. We conducted a structural analysis of pyridylaminated N-linked oligosaccharides prepared from galactomannoproteins by HPLC and 1H NMR. These analyses revealed that the N-linked oligosaccharides of the och1δ cells displayed heterogeneity in the glycan consisting of Hex11-15GlcNAc2. The structural heterogeneity arose mainly from the addition of γ1,2- and γ1,3-Gal residues to the Man9GlcNAc2 core structure.

Original languageEnglish
Pages (from-to)407-414
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number2
DOIs
Publication statusPublished - May 7 2009

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Schizosaccharomyces
Oligosaccharides
Yeast
Structural analysis
Mannosyltransferases
Saccharomycetales
Membrane Glycoproteins
Substrate Specificity
Galactose
Lectins
Polysaccharides
Saccharomyces cerevisiae
Elongation
Genes
High Pressure Liquid Chromatography
Nuclear magnetic resonance
Substrates
Enzymes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry
  • Medicine(all)

Cite this

The och1 mutant of schizosaccharomyces pombe produces galactosylated core structures of n-linked oligosaccharides. / Ohashi, Takao; Ikeda, Yuka; Tanaka, Naotaka; Nakakita, Shin Ichi; Natsuka, Shunji; Giga-Hama, Yuko; Takegawa, Kaoru.

In: Bioscience, Biotechnology and Biochemistry, Vol. 73, No. 2, 07.05.2009, p. 407-414.

Research output: Contribution to journalArticle

Ohashi, Takao ; Ikeda, Yuka ; Tanaka, Naotaka ; Nakakita, Shin Ichi ; Natsuka, Shunji ; Giga-Hama, Yuko ; Takegawa, Kaoru. / The och1 mutant of schizosaccharomyces pombe produces galactosylated core structures of n-linked oligosaccharides. In: Bioscience, Biotechnology and Biochemistry. 2009 ; Vol. 73, No. 2. pp. 407-414.
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