Structural Analysis of N-Linked Oligosaccharide of Mitogenic Lectin-B from the Roots of Pokeweed (Phytolacca americana)

Yoshinobu Kimura, Ken Ichi Yamaguchi, Gunki Funatsu

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Abstract

The structure of an asparagine (N-) linked oligosaccharide of pokeweed lectin-B (PL-B) and the amino acid sequences around two glycosylation sites were identified. The pyridylamino (PA) oligosaccharide prepared from PL-B was eluted as a single peak on both reversed-phase (RP-) HPLC and size-fractionation (SF-) HPLC, and its structure was estimated to be Manal →6(Manαl → 3)(Xylβ1→2) Manβ1→4GlcNAcβ1β4(Fucα1→3)GlcNAc by a combination of component analysis, successive exoglycosidase digestions, IS-MS analysis, and 500 MHz 1H-NMR. Two tryptic glycopeptides were isolated from the reduced and S-pyridylethylated PL-B after gel filtration followed by RP-HPLC, indicating the presence of two glycosylation sites in PL-B. The amino acid sequences around the two glycosylation sites were determined to be Cys-Gly-Val-Asp-Phe-Gly-Asn(CHO)-Arg.

Original languageEnglish
Pages (from-to)537-540
Number of pages4
JournalBioscience, Biotechnology and Biochemistry
Volume60
Issue number3
Publication statusPublished - Mar 1996

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Keywords

  • IS-MS
  • Mitogenic lectin
  • N-linked oligosaccharide
  • Phytolacca americana
  • Plant glycoprotein

ASJC Scopus subject areas

  • Food Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Bioengineering

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