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Glycome analysis by tandem two-dimensional chromatography

Research Project

Project/Area Number 15590038
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionChiba University

Principal Investigator

TOYODA Hidenao  Chiba University, Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学研究院, 助教授 (70217579)

Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsChromatography / Glycan / Glycosaminoglycan / Fluorescence / 高速液体クロマトグラフィー / ムチン型糖鎖
Research Abstract

We have devised a method for glycome analysis by using on-line two-dimensional HPLC. N-linked oligosaccharides, mucins and glycosaminoglycans were separated simultaneously by the HPLC. We applied this method to a genetically tractable model organism, Drosophila melanogaster. In addition to this chemical analysis, western-blot analysis by use of lectin staining and immunostaining provides attractive tools to further understanding of basic physiological functions of glycoconjugates. Thus, we carried out glycoconjugate profiling of Drosophila by 1D or 2D-electrophoresis with western-blot analysis followed lectin staining and immunostaining. Drosophila glycoconjugates were well characterized by SBA, ConA and WGA lectins and the method was applied to flies bearing mutations in glyco-genes. Furthermore, proteoglycans were profiled by western-blot analysis with immunostaining. These methods are planning to be employed in the proteo-glycomics study.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (10 results)

All 2005 2004 2003 Other

All Journal Article (7 results) Publications (3 results)

  • [Journal Article] Structural characterization of human liver heparan sulfate2005

    • Author(s)
      P.Vongchan, M.Warda, H.Toyoda, T.Toida, R.M.Marks, R.J.Linhardt
    • Journal Title

      Biochimica et Biophysica Acta 1721

      Pages: 1-8

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Approach for functional analysis of glycan using RNA interference2004

    • Author(s)
      S.Nishihara, R.Ueda, S.Goto, H.Toyoda, H.Ishida, M.Nakamura
    • Journal Title

      Glycoconjugate Journal 21

      Pages: 63-68

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorihabditis elegans2003

    • Author(s)
      H.Morio, Y.Honda, H.Toyoda, M.Nakajima, H.Kurosawa, T.Shirasawa
    • Journal Title

      Biochemical and Biophysical Research Communications 301

      Pages: 317-323

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter2003

    • Author(s)
      S.Kamiyama, T.Suda, R.Ueda, M.Suzuki, R.Okubo, N.Kikuchi, Y.Chiba, S.Goto, H.Toyoda, K.Saigo, M.Watanabe, H.Narimatsu, Y.Jigami, S.Nishihara
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 25958-25963

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate2003

    • Author(s)
      H.Barth, C.Shafer, M.I.Adah, F.Zhang, R.J.Linhardt, H.Toyoda, A.Kinoshita-Toyoda, T.Toida, T.H.Van Kuppevelt, E.Depla, F.Von Weizsacker, H.E.Blum, T.F.Baumert
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 41003-41012

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorhabditis elegans2003

    • Author(s)
      H.Morio, Y.Honda, H.Toyoda, M.Nakajima, H.Kurosawa, T.Shirasawa
    • Journal Title

      Biochemical and Biophysical Research Communications 301

      Pages: 317-323

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate2003

    • Author(s)
      H.Barth, C.Schafer, M.I.Adah, F.Zhang, R.J.Linhardt, H.Toyoda, A.Kinoshita-Toyoda, T.Toida, T.H.Van Kuppevelt, E.Depla, B.F.Von Weizsacker, H.E.Blum, T.F.Baumert
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 41003-41012

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] H.Morio, Y.Honda, H.Toyoda, M.Nakajima, H.Kurosawa, T.Shirasawa: "EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorhabditis elegans"Biochemical and Biophysical Research Communications. 301. 317-323 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] S.Kamiyama, T.Suda, R.Ueda, M.Suzuki, R.Okubo, N.Kikuchi, Y.Chiba, S.Goto, H.Toyoda, K.Saigo, M.Watanabe, H.Narimatsu, Y.Jigami, S.Nishihara: "Molecular cloning and identification of 3´-phosphoadenosine 5´-phosphosulfate transporter"Journal of Biological Chemistry. 278. 25958-25963 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Barth, C.Schafer, M.I.Adah, F.Zhang, R.J.Linhardt, H.Toyoda, A.Kinoshita-Toyoda, T.Toida, T.H.Van Kuppevelt, E.Depla, F.Von Weizsacker, H.E.Blum, T.F.Baumert: "Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate"Journal of Biological Chemistry. 278. 41003-41012 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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