Glycome analysis by tandem two-dimensional chromatography

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 15590038
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Chiba 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
• Keywords: Chromatography / Glycan / Glycosaminoglycan / Fluorescence

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.

Research Products

• [Journal Article] Structural characterization of human liver heparan sulfate (2005)
  • 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
• [Journal Article] Approach for functional analysis of glycan using RNA interference (2004)
  • 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
• [Journal Article] EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorihabditis elegans (2003)
  • 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
• [Journal Article] Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter (2003)
  • 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
• [Journal Article] Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate (2003)
  • 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
• [Journal Article] EXT gene family member rib-2 is essential for embryonic development and heparan sulfate biosynthesis in Caenorhabditis elegans (2003)
  • 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
• [Journal Article] Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate (2003)
  • 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