Structural features of n-glycans linked to royal jelly glycoproteins: Structures of high-mannose type, hybrid type, and biantennary type glycans

Yoshinobu Kimura, Chiyoko Miyagi, Mariko Kimura, Teruhiko Nitoda, Nobuyuki Kawai, Hiroyuki Sugimoto

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51 Citations (Scopus)


The structures of N-glycans of total glycoproteins in royal jelly have been explored to clarify whether antigenic N-glycans occur in the famous health food. The structural feature of N-glycans linked to glycoproteins in royal jelly was first characterized by immunoblotting with an antiserum against plant complex type N-glycan and lectin-blotting with Con A and WGA. For the detail structural analysis of such N-glycans, the pyridylaminated (PA-) N-glycans were prepared from hydrazinolysates of total glycoproteins in royal jelly and each PA-sugar chain was purified by reverse-phase HPLC and size-fractionation HPLC. Each structure of the PA-sugar chains purified was identified by the combination of two-dimensional PA-sugar chain mapping, ESI-MS and MS/MS analyses, sequential exoglycosidase digestions, and 500 MHz 1H-NMR spectrometry. The immunoblotting and lectinblotting analyses preliminarily suggested the absence of antigenic N-glycan bearing β1-2 xylosyl and/or α1-3 fucosyl residue(s) and occurrence of β1-4GlcNAc residue in the insect glycoproteins. The detailed structural analysis of N-glycans of total royal jelly glycoproteins revealed that the antigenic N-glycans do not occur but the typical high mannose-type structure (Man9-4GlcNAc2) occupies 71.6% of total N-glycan, biantennary-type structures (GlcNAc2Man3GlcNAc2) 8.4%, and hybrid type structure (GlcNAc1Man4GlcNAc2) 3.0%. Although the complete structures of the remaining 17% N-glycans; C4, (HexNAc3Hex3HexNAc2: 3.0%), D2 (HexNAc2Hex5HexNAc2: 4.5%), and D3 (HexNAc3Hex4HexNAc2: 9.5%) are still obscure so far, ESI-MS analysis, exoglycosidase digestions by two kinds of β-N-acetylglucosaminidase, and WGA blotting suggested that these N-glycans might bear a β1-4 linkage N-acetylglucosaminyl residue.

Original languageEnglish
Pages (from-to)2109-2120
Number of pages12
JournalBioscience, Biotechnology and Biochemistry
Issue number10
Publication statusPublished - 2000


  • Antigenic N-glycan
  • Insect glycoprotein
  • N-glycan glycoform
  • Royal jelly

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry


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