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

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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
Publication statusPublished - Nov 1996

Fingerprint

Peas
Tandem Mass Spectrometry
Sugars
Pisum sativum
Mass spectrometry
Seed
glycoproteins
peas
Seeds
Glycoproteins
High Pressure Liquid Chromatography
Ions
ions
sugars
Acetylglucosaminidase
Glycoside Hydrolases
Xylose
seeds
Digestion
glycosidases

Keywords

  • endo-β-N-acetylglucosaminidase- PS
  • IS-MS/MS
  • N-linked sugar chain
  • Pisum sativum
  • plant glycoprotein

ASJC Scopus subject areas

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

Cite this

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title = "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)",
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.",
keywords = "endo-β-N-acetylglucosaminidase- PS, IS-MS/MS, N-linked sugar chain, Pisum sativum, plant glycoprotein",
author = "Yoshinobu Kimura and Akira Ohno and Shigeaki Takagi",
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T1 - 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)

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AU - Ohno, Akira

AU - Takagi, Shigeaki

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N2 - 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.

AB - 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.

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