Demonstration of the presence of α-1,6-branched side chains in the mannan of Candida stellatoidea

Nobuyuki Shibata, Nobutaka Senbongi, Tomoko Hosoya, Kumi Kawahara, Rieko Akagi, Akifumi Suzuki, Hidemitsu Kobayashi, Shigeo Suzuki, Yoshio Okawa

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9 Citations (Scopus)

Abstract

A mild acetolysis of the mannans of Candida stellatoidea was performed after acetylation to yielded an α-1,6-branched mannohexaose, the presence of which had been predicted from the appearance of a specific H1-H2-correlated cross-peak in two-dimensional homonuclear Hartmann-Hahn spectroscopy. In this study, we found that the de-O-acetylation of a 4-O-acetyl group at the branching point, the 3,6-di-O-substituted mannose unit, of an acetylated oligosaccharide by sodium methoxide is significantly slower than that of other acetyl groups. We could separate the 4-O-acetylated branching oligosaccharide from linear isomer using high-performance liquid chromatography. Before and after the de-O-acetylation of the purified branching oligosaccharide, their 1H-NMR signals were sequentially assigned by means of the nuclear Overhauser effect. In the sequential NMR assignment study, we showed that the α-1,6-linked mannose unit is attached to the 3-O-substituted unit based on the presence of NOE cross-peak between H1 of the branching mannose unit and 1-16 of the 3-O-substituted mannose unit. An enzyme-linked immunosorbent inhibition assay of the reactivity of factor 4 serum to C. stellataoidea mannan by several oligosaccharides indicated that the α-1,6-branched oligosaccharide and the β-1,2 linkage-containing oligosaccharides showed inhibitory activity. This result indicates that factor 4 serum, as well as factor 5 and 6 sera, contains antibodies against β-1,2 linked mannose units which have been reported to participate in pathogenicity via cytokine production and/or adherence. From the assignment results of H1-H2-correlated cross-peaks of oligosaccharides and mannans, the molar ratio of the mannan side chains was proposed. In this study, we demonstrated that the epitope structure of the C. stellatoidea type I strains was the same as that of the C. albicans NIH B-792 (serotype B) strain.

Original languageEnglish
Pages (from-to)477-485
Number of pages9
JournalEuropean Journal of Biochemistry
Volume246
Issue number2
DOIs
Publication statusPublished - 1997 Jan 1
Externally publishedYes

Keywords

  • Assignment
  • Candida stellatoidea
  • Mannan
  • Mannooligosaccharide
  • NMR

ASJC Scopus subject areas

  • Biochemistry

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    Shibata, N., Senbongi, N., Hosoya, T., Kawahara, K., Akagi, R., Suzuki, A., Kobayashi, H., Suzuki, S., & Okawa, Y. (1997). Demonstration of the presence of α-1,6-branched side chains in the mannan of Candida stellatoidea. European Journal of Biochemistry, 246(2), 477-485. https://doi.org/10.1111/j.1432-1033.1997.00477.x