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1999 Fiscal Year Final Research Report Summary

Analysis of Oligosaccharide Structures from the Reducing End Terminal by Combining Partial Acid Hydrolysis and a Two-Dimensional Sugar Map

Research Project

Project/Area Number 10558100
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

HASE Sumihiro  Department of Chemistry, Graduate School of Science, Osaka University, 大学院・理学研究科, 教授 (80028232)

Co-Investigator(Kenkyū-buntansha) TSUKAMOTO Shigehisa  Analytical Chemistry Research Laboratory, IRICA Instruments, Inc. Electronic engineer, 開発研究部, 研究員
Project Period (FY) 1998 – 1999
KeywordsSugar chain / 2D-map / Pyridylaminotion / oligosaccharide / partial acid hydrolysis / Sturcfure
Research Abstract

Structure analysis is a necessary step in elucidating the roles of glycoprotein sugar chains. Glycoproteins with biological activity isolated from natural sources, however, are often obtainable in only limited amounts. To circumvent this drawback, we developed a fluorescence labeling method, which we have been using in combination with exoglycosidase digestion. Although the method is useful, we frequently encounter difficulty in obtaining enzymes or in the incubation conditions, and even when we do have an enzyme, it is not always able to liberate sugar residues due to substrate specificity.
To overcome this, a new sensitive and convenient method for the structural analysis of oligosaccharides was developed, in which partial acid hydrolysis was combined with two-dimensional sugar mapping of pyridylamino (PA-) oligosaccharides. APA-oligosaccharide was partially hydrolyzed, the acid hydrolysis conditions being optimized so as to obtain various PA-olgosaccharide fragments with high yields from different types of PA-oligosaccharides. The acid hydrolyzates were then fractionated every 1 glucose unit by size-fractionation HPLC, and each fraction was analyzed by reversed-phase HPLC. The structure of each PA-oligosaccharide fragment was identified on a two-dimensional map prepared with standard PA-sugar chains, after which the original PA-oligosaccharide was reconstructed from the reducing end terminal based on the obtained structures of the PA-oligosaccharide fragments. The method was successfully applied to resolving the structure of an N-linked sugar chain.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] A. Shimazaki, et al.: "A New Suger Chain of the Proteinase Inhibitor from Latex of Carica papaya"J. Biochem.. 125・3. 560-565 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H. Wakabayashi, et al.: "Novel Proteoglycan Linkage Tetrasaccharides of Human Urinary Soluble Thrombomodulin, SO_4-3GlcAβ1-3Galβ1-3(±Sia α2-6)Galβ1-4Xyl"J. Biol. Chem.. 274・9. 5436-5442 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K. yanagida, et al.: "A Pyridylamination method aimed at automatic oligosaccharide Analysis of N-linked suger chains"Anal. Biochem.. 274・2. 229-234 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuma Iwai: "Detection of Man5GlcNAc and Related Free oligosaccharides in the Cytosol Fraction of Hen Oviduct"J. Biochem.. 125・1. 70-74 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kanta yanagida: "Introduction of a New Scale Into Reversed-phase High-performance Liquid Chromatography of Pyridylamino Suger Chains for Structural Assignment"J. Chromatogr. A. 800. 187-198 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasushi Makino: "Analysis of Oligosaccharide Structures from the Reducing End Terminal by Combining Partial Acid Hydrolysis and a Two-dimensional Suger Map"Anal. Biochem.. 264・2. 172-179 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shunji Natsuka: "Analysis of N- and O-Glycans by Pyridylamination"Humana Press. 13 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 長谷純宏: "生産と技術誌51巻"蛍光標識法による糖鎖の構造と機能の解析. 4 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y. Makino, Kaoru Omichi and S. Hase: "Analysis of Oligosaccharide Structures from the Reducing End Terminal by Combining Partial Acid Hydrolysis and a Two-Dimensional Sugar Map."Anal. Biochem.. 264, (2). 172-179 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Yanagida, H. Ogawa, K. Omichi and S. Hase: "Introduction of a New Scale into Reversed-Performance Liquid Chromatography of Pyridylamino Sugar Chains for Structural Assigment."J. Chromatogr. A. 800. 187-198 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Shimazaki, Y. Makino, K. Omichi, S. Odani and S. Hase: "A New Sugar Chain of the Proteinase Inhibitor from Latex of Carica papaya."J. Biochem.. 125. 560-565 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Wakabayashi, S. Natsuka, T. Mega, N. Otsuki, M. Isaji, Masaki Naotsuka, S. Koyama, T. Kanamori, K. Sakai and S. Hase: "Novel Proteoglycan Linkage Tetrasaccharides of Human Urinary Soluble Thrombomoduin, SOィイD24ィエD2-3GlcAβ1-3Galβ1-3(±Siaα2-6)Galβ1-4Xyl."J. Biol. Chem.. 274, (9). 5436-5442 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Yanagida, S. Natsuka, and S. Hase: "A pyridylamination method aimed at automatic oligosaccharide Analysis of N-linked sugar chains."Anal. Biochem.. 274. 229-234 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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