Structural elucidation of n-linked sugar chains of storage glycoproteins in mature pea (pisum sativum) seeds by ion-spray tandem mass spectrometry (IS-MS/MS)

Yoshinobu Kimura, Akira Ohno, Shigeaki Takagi

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The structures of N-linked sugar chains of the storage glycoproteins in mature pea seeds have been estimated. Nine pyridylaminated (PA-) N-linked sugar chains were derived and purified from the hydrazinolysate of the storage glycoproteins by reversed-phase HPLC and size-fractionation HPLC. The structures of the PA-sugar chains purified were first identified by two-dimensional PA-sugar chain mapping, considering the data of sugar composition analysis or sequential exoglycosidase digestions. The deduced structures were further analyzed by IS-MS/MS analysis. Every relevant fragment ion derived from all PA-sugar chains could be assigned on the basis of deduced structures. The estimated nine structures fell into two categories; the first was a typical oligomannose type (Man8-3GlcNAc2; 77.7%) which can be hydrolyzed by endo-β-N-acetylglucosaminidase PS [Y. Kimura et al., Biosci. Biotech. Biochem., 60, 228–232 (1996)], the second was a xylose-containing type (Man4-3Xyl1GlcNAc2, Man3Fuc1Xyl1GlcNAc2; 22.3%). Among these structures, Man8GlcNAc2 (19.7%), Man6GlcNAc2 (24.7%), and Man3Fuc1Xyl1GlcNAc2 (18.8%) were the dominant structures.

Original languageEnglish
Pages (from-to)1841-1850
Number of pages10
JournalBioscience, Biotechnology and Biochemistry
Volume60
Issue number11
DOIs
Publication statusPublished - Jan 1996

Keywords

  • Endo-β-N-acetylglucosaminidase-PS
  • IS-MS/MS
  • N-linked sugar chain
  • Pisum sativum
  • Plant glycoprotein

ASJC Scopus subject areas

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

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