Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells: Predominant occurrence of complex type N-glycans

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Abstract

Although it has been found that plant endo-β-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-β-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man 9-5GlcNAc1) occur at concentration of several micromolar (μM). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.

Original languageEnglish
Pages (from-to)1356-1363
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume70
Issue number6
DOIs
Publication statusPublished - 2006

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Glycoproteins
Cell culture
Polysaccharides
glycoproteins
cell culture
polysaccharides
Cell Culture Techniques
Mannose
mannose
rice
Bearings (structural)
Epitopes
Acetylglucosaminidase
Substrates
Structural analysis
Genes
glycopeptides
Plant Structures
Glycopeptides
glycosidases

Keywords

  • N-glycan metabolism
  • N-glycan structures
  • Oriza sativa
  • Rice glycoprotein

ASJC Scopus subject areas

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

Cite this

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title = "Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells: Predominant occurrence of complex type N-glycans",
abstract = "Although it has been found that plant endo-β-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-β-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man 9-5GlcNAc1) occur at concentration of several micromolar (μM). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95{\%} of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5{\%} of total N-glycans.",
keywords = "N-glycan metabolism, N-glycan structures, Oriza sativa, Rice glycoprotein",
author = "Megumi Maeda and Yoshinobu Kimura",
year = "2006",
doi = "10.1271/bbb.50637",
language = "English",
volume = "70",
pages = "1356--1363",
journal = "Bioscience, Biotechnology and Biochemistry",
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T1 - Glycoform analysis of N-glycans linked to glycoproteins expressed in rice culture cells

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AU - Maeda, Megumi

AU - Kimura, Yoshinobu

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AB - Although it has been found that plant endo-β-N-acetylglucosaminidase shows strong activity towards denatured glycoproteins and glycopeptides with high-mannose type N-glycans and free high-mannose type N-glycans bearing the chitobiosyl unit, the endogenous substrates for plant endoglycosidase have not yet been identified. Recently we purified and characterized an endo-β-N-acetylglucosaminidase from rice culture cells and identified the gene encoded (Maeda, M., and Kimura, Y., Trends Glycosci. Glycotech., 17, 205-214 (2005)). Furthermore, we found structural features of free N-glycans in the cells, indicating that high-mannose type species (Man 9-5GlcNAc1) occur at concentration of several micromolar (μM). Hence, in this study we analyzed glycoform of N-glycans linked to glycoproteins expressed in rice culture cells to see whether endogenous glycoproteinous substrate occurs in reasonable amounts. Structural analysis revealed that more than 95% of total N-glycans linked to glycoproteins in the rice cells had the plant complex type structure, including Lewis a epitope-harboring type, although high-mannose type structures account for less than 5% of total N-glycans.

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KW - N-glycan structures

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KW - Rice glycoprotein

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