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Development of methods to determine carbohydrate structure by Fourier transform mass spectrometry

Research Project

Project/Area Number 09558083
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Structural biochemistry
Research InstitutionTokyo Metropolitan Institute of Gerontology

Principal Investigator

ENDO Tamao  Tokyo Metropolitan Institute of Gerontology, Department of Glycobiology, Head, 糖鎖生物学部門, 研究室長 (30168827)

Co-Investigator(Kenkyū-buntansha) SASAKI Tasuku  Tokyo Metropolitan Institute of Gerontology, Department of Glycobiology, Researcher, 糖鎖生物学部門, 研究員 (40291132)
SATO Yuji  Tokyo Metropolitan Institute of Gerontology, Department of Glycobiology, Researcher, 糖鎖生物学部門, 研究員 (90280768)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 1999: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1998: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1997: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsFourier transform mass spectrometry (FTMS) / Glycoprotein / Carbohydrate / post source decay / MALDI-TOM / MS / 2-Aminobenzamide / 蛍光標識
Research Abstract

Molecular weight of seven 2-aminobenzamide (2AB)-derivatized oligosaccharides could be determined as small as 10 pmol by matrix-assisted laser desorption ionization (MALDI)-Fourier transform mass spectrometry (FTMS) with a good signal-to-noise ratio. Interestingly, all ions from either high-mannose or complex-type sugar chains have shown spontaneously with the release of GIcNac-3AB by B-type cleavage. Additionally, loss of two hexoses was found from nonreducing terminal from bi-, tri-, and tetra-antennary complex-type sugars, suggesting that galactose residues on GIcNAcβ1-2Man arm may be deleted specifically. On the other hand, ManィイD29ィエD2GIcNAcィイD22ィエD2-2AB released three hexoses from nonreducing termini, suggesting three Manα1-2 linkages may be fragile. 2AB-derivatized oligosaccharides were separated by three lectin column chromatographies and then subjected to MALDI time-of-flight mass spectrometry (MALDI-TOF/MS) for structural characterization of the carbohydrates. The combination … More of sequential exoglyconsidase digestion and MALDI-TOF/MS greatly facilitates the monosaccharide sequencing and is more feasible than size-exclusion column chromatography in terms of the time consumed and the laboriousness of the procedure. By this strategy, microsequencing of 2-3 pmol of oligosaccharide derivatives could be achieved. Furthermore, spectra obtained by the post source decay (PSD) mode provide excellent sequence information. The relative intensities of metastable ions due to fragmentation at glycosidic linkages were different among linkage isomers of particular oligosaccharides. These results demonstrate that PSD analysis possesses significant potential for the estimation of glycosidic linkage in carbohydrate structures. The quantitative nature of MALDI-TOF/MS in oligosaccharide analysis was of great use in the comparative study on normal and pathogenic prion proteins, PrPィイD1cィエD1 and PrPィイD1ScィエD1, respectively. Our developed methods enable analysis of glycan mixtures quantitatively. Less

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Yuji Sato: "Microsequencing of glycans using 2-aminobenzamide and MALDI-TOF mass spectrometry : occurrence of unique linkage-dependent fragmentation"Anal. Chem.. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tamao Endo: "O-Mannosyl glycan in mammals"Biochim. Biophys. Acta.. 1473. 237-246 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Sato: "Study of the sugar chains of recombinant human amyloid precursor protein produced by Chinese hamster ovary cells"Biochim. Biophys. Acta.. 1472. 344-358 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Megumi T-Hiruma: "Detection of novel carbohydrate binding activity of interleukin-1"J. Biol . Chem.. 274. 4459-4466 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Puline M.Rudd: "Glycosylation differences between the normal and pathogenic prion protein"Proc. Natl. Acad. Sci. USA. 96. 13044-13049 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tasuku Sasaki: "Detection of O-mannosyl glycans in rabbit skeletal muscle α-dystroglycan"Biochem. Biophys. Acta. 1425. 599-606 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tamao Endo: "Sialobiology and Other Novel Forms of Glycosylation"Gakushin Publisher. 5 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 遠藤玉夫: "糖鎖生物学"共立出版. 7 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Sato: "Microsequencing of glycans using 2-aminobenzamide and MLDI-TOF mass spectrometry Occurrence of unique linkage-dependent fragmentation"Anal. Chem.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Pauline Rudd: "Glycosylation differences between the normal and pathogenic prion protein isoforms"Proc. Natl. Acad. Sci. USA.. 96. 13044-13049 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tamao Endo: "O-Mannosyl glycan in mammals"Biochem. Biophys. Acta. 1473. 237-246 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Megumi Tandai-Hiruma: "Detection of novel carbohydrate binding activity of interleukin-1"J. Biol. Chem.,. 274. 4459-4466 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tasuku Sasaki: "Detection of O-mannosyl glycans in rabbit skeletal muscle α-dystroglycan"Biochem. Biophys. Acta. 1425. 599-606 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Sato: "Microsequencing of glycans using 2-aminobenzamide and MALDI-TOF mass spectrometry : occurrence of unique linkage-dependent fragmentation"Anal. Chem.. (印刷中).

    • Related Report
      1999 Annual Research Report
  • [Publications] Tamao Endo: "O-Mannosyl glycan in mammals"Biochim. Biophys. Acta.. 1473. 237-246 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Megumi T-Hiruma: "Detection of novel carbohydrate binding activity of interleukin-1"J. Biol. Chem.. 274. 4459-4466 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Pauline M. Rudd: "Glycosylation differences between the normal and pathogenic prion protein"Proc. Natl. Acad. Sci. USA. 96. 13044-13049 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tamao Endo: "Sialobiology and Other Novel Forms of Glycosylation"Gakushin Publisher. 5 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yuji Sato: "Comparison of lectin-binding patterns between young adult and older rat glycoproteins in the brain" Glycoconjugate J.(印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Yuji Sato: "Differential expression of sialoglycoproteins in the rat hippocampus and its changes during aging" Neuroscience Lett.(印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Yuji Sato: "Evidence for the presence of major peripheral myelin glycoprotein P_o in mammalian spinal cord and a change of its glycoprotein state during aging" Glycobiology. (印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Megumi T-Hiruma: "Detection of novel carbohydrate binding activity of interleukin-1" J.Biol.Chem.(印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] 遠藤玉夫: "糖鎖生物学" 共立出版, 7 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 遠藤玉夫: "Annual Review 神経 1998" 中外医学社, 8 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tamao Endo: "Structural differences among serum IgA proteins of chimpanzee,Rhesus monkey and rat orgin" Molec.Immunol.34・7. 557-565 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kiitiro Matsumura: "A role of dystroglycan in schwannoma cell adhesion to laminin" J.Biol.Chem.272・21. 13904-13910 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoyuki Tsuchiya: "Lectin Methods and Protocols" Humana Press, 11 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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