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2007 Fiscal Year Final Research Report Summary

Function search of glycoconjugates by selective laser desorption ionization mass spectronmeter using metal nanoparticle

Research Project

Project/Area Number 17205015
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemistry related to living body
Research InstitutionHokkaido University

Principal Investigator

NISIMURA Shin-ichitro  Hokkaido University, Graduate School of Advanced Life Science, Professor (00183898)

Co-Investigator(Kenkyū-buntansha) NAGAHORI Noriko  Hokkaido University, Graduate School of Advanced Life Science, Assistant Professor (90372268)
HINOU Hiroshi  Hckkaido University, Graduate School of Advanced Life Science, Assistant Professor (70333333)
Project Period (FY) 2005 – 2007
Keywordsgold colloidal nanoparticle / carbohydrate / glycoblotting / suicide substrate / MALDI-TOF MS / glycomics / quantum dots / live cell imaging
Research Abstract

this research was performed to understand the functions of glycoconjugates at the molecular level, that is to say, to develop a new methodology for hunting and identifying the partner molecules with target carbohydrates by using the gold colloidal nanoparticle and the mass spectrometer.
<Preparation water -soluble and non-specific adsorption depression type gold colloidal nanoparticle displaying carbohydrates>
in this study, the modification of nanoparticles was performed using specific affinity with a metallic surface and thiol group. Strong interaction of carbohydrate-displaying nanoparticle with a partner molecule is expected, because several hundred of carbohydrate molecules are displayed onto a surface of nanoparticle (carbohydrate cluster effect). In addition, the system that could display any carbohydrates was built by using chemoselective reaction with aminooxy1 group and a reducing end of carbohydrate. A library of carbohydrate presentation nano particle was made using this syst … More em. In addition, the surfaces of the nanoparticles were modified with suicide substrate type carbohydrate derivatives that could anchor the target proteins with covalent bonds. The presentation of carbohydrates on the nanoparticles was evaluated by MALDI-TOF MS using that the laser cleaved metal-thiol bond. In order to reduce the non-specific adhesion of proteins, a thiol compound with zwitterionic head group was designed and synthesized. All carbohydrate-displaying metal nanoparticles prepared in this study showed high water solubility and stability.
<Establishment of the methods for hunting and identifying the partner molecules of carbohydrates>
The highly water-soluble nanoparticles could be directly subjected to a various biochemical assays and MALDI-TOF MS measurement after the capture of partner molecules. Incubate nanoparticle and sample solution containing partner molecules, and wash the particle with buffer, then the particle was just applied to SDS-PAGE. The protein identification was possible according to the ordinary method of proteomics. In this way, a high sensitivity by reduced sample loss and a promotion of efficiency by simplification of activity process were achieved, compared with traditional approaches such as the affinity chromatography which needed elution by a competing carbohydrate. Less

  • Research Products

    (27 results)

All 2008 2007 2006 2005

All Journal Article (17 results) (of which Peer Reviewed: 7 results) Presentation (2 results) Book (5 results) Patent(Industrial Property Rights) (3 results) (of which Overseas: 1 results)

  • [Journal Article] A comprehensive approach to structural and functional glycomics based on chemoselective glycoblotting and sequential tag conversion2008

    • Author(s)
      Furukawa, J-i.
    • Journal Title

      Anal. Chem. 4

      Pages: 1094-1101

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] BlotGlycoABCTM: An integrated glycoblotting technique for rapid and large-scale clinical glycomics2008

    • Author(s)
      Miura, Y.
    • Journal Title

      Mol. Cell. Proteom 7

      Pages: 370-377

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Reverse glycoblotting allows rapid enrichment glycoproteomics of biopharmaceuticals and disease-related biomarkers2008

    • Author(s)
      Kurogochi, M.
    • Journal Title

      Angew. Chem. Int. Ed. 46

      Pages: 8808-8813

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] A comprehensive approach to structural and functional glycomics based on chemoselective glycoblotting and sequential tag conversion2008

    • Author(s)
      Furukawa J-i., Shinohara Y., Kuramoto H., Miura Y., Shimaoka H., Kurogochi M., Nakano M., Nishimura S.-I.
    • Journal Title

      Anal. Chem. 4

      Pages: 1094-1101

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] BlotGlycoABCTM: An integrated glycoblotting technique for rapid and large-scale clinical glycomics2008

    • Author(s)
      Miura Y., Hato M., Shinohara Y., Kuramoto H., Furukawa J.-i., Kurogochi M., Shimaoka H., Tada M., Nakanishi K., Ozaki M., Todo S., Nishimura S.-I.
    • Journal Title

      Mol. Cell. Proteom. 7

      Pages: 370-377

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Reverse glycoblotting allows rapid enrichment glycoproteomics of biopharmaceuticals and disease-related biomarkers2008

    • Author(s)
      Kurogochi M., Amano M., Fumoto M., Takimoto A., Kondo H., Nishimura S.-I.
    • Journal Title

      Angew. Chem. Int. Ed. 46

      Pages: 8808-8813

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Rapid and quantitative solid-phase esterification of sialic acid residues permits high throughput and reliable glycomics by mass spectrometry2007

    • Author(s)
      Miura, Y.
    • Journal Title

      Chem. Eur. J. 13

      Pages: 4797-4804

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Rapid and Simple Solid-Phase Esterification of Sialic Acid Residues for Quantitative Glycomics by Mass Spectrometry (pNA)2007

    • Author(s)
      Miura Y., Shinohara Y., Furukawa J., Nagahori N., Nishimura S.-I.
    • Journal Title

      Chem. Eur. J. 13

      Pages: 4797-4804

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] D: Molecular interactions: Multivalent presentation: Glycopolymers2007

    • Author(s)
      Nagahori N., Nishimura S.-I.
    • Journal Title

      Comprehensive Glycoscience (Elsevier Ltd.)

      Pages: 453-475

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Direct and efficient monitoring of glycosyltransferase reaction on gold colloidal nanoparticles by mass spectrometry2006

    • Author(s)
      N, Nagahori
    • Journal Title

      Chem. Eur. J. 12

      Pages: 6478-6485

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and sphingoglycolipids2006

    • Author(s)
      Naruchi, K.
    • Journal Title

      J. Org. Chem. 71

      Pages: 9609-9621

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Unusual N-Glycan Structures in α-Mannosidase II/IIx Double Null Embryos Identified by a Systematic Glycomics Approach Based on Two-dimensional LC Mapping and Matrix-dependent Selective Fragmentation Method in MALDI-TOF/TOF Mass Spectrometry2006

    • Author(s)
      Hato, M.
    • Journal Title

      Mol. Cell. Proteom. 5

      Pages: 2146-2157

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Direct and Efficient Monitoring of Glycosyltransferase Reactions on Gold Colloidal Nanoparticles by Using Mass Spectrometry2006

    • Author(s)
      Nagahori N., Nishimura S.-I.
    • Journal Title

      Chem. Eur. J. 12

      Pages: 6478-6485

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and sphingoglycolipids2006

    • Author(s)
      Naruchi K., Hamamoto T., Kurogochi M., Hinou H., Shimizu H., Matsushita T., Fujitani N., Kondo H., Nishimura S.-I.
    • Journal Title

      J. Org. Chem. 71

      Pages: 9609-9621

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Unusual N-Glycan Structures in α-Mannosidase II/IIx Double Null Embryos Identified by a Systematic Glycomics Approach Based on Two-dimensional LC Mapping and Matrix-dependent Selective Fragmentation Method in MALDI-TOF/TOF Mass Spectrometry2006

    • Author(s)
      Hato M., Nakagawa H., Kurogochi M., Akama T. O., Marth J. D., Fukuda M. N., Nishimura S.-I.
    • Journal Title

      Mol. Cell. Proteom. 5

      Pages: 2146-2157

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Mechanism-Based Inhibitors to Probe Transitional States of Glycoside Hydrolases2006

    • Author(s)
      Hinou H., Kurogochi M., Nishimura S.-I.
    • Journal Title

      Methods in Enzymology (Academic Press) 415

      Pages: 202-212

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Glyconanotecnology2006

    • Author(s)
      Nagahori N., Nishimura S.-I.
    • Journal Title

      Nanotechnology in Carbohydrate Chemistry (Yuasa H., Transworld Research Network)

      Pages: 149-166

    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] 糖脂質レプリカナノ微粒子2007

    • Author(s)
      長堀, 紀子
    • Organizer
      日本糖質学会
    • Place of Presentation
      九州大学
    • Year and Date
      2007-08-03
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] GSLs replica nanoparticle2007

    • Author(s)
      Nagahori, N.
    • Organizer
      The Japanese Society of Carbohydrate Research
    • Place of Presentation
      Kyushu University
    • Year and Date
      2007-08-03
    • Description
      「研究成果報告書概要(欧文)」より
  • [Book] ケミカルバイオロジー、蛋白質核酸酵素 vo1522007

    • Author(s)
      長堀, 紀子
    • Total Pages
      8
    • Publisher
      共立出版
    • Description
      「研究成果報告書概要(和文)」より
  • [Book] Comprehensive Glycoscience2007

    • Author(s)
      N., Nagahori
    • Total Pages
      23
    • Publisher
      Elsevier
    • Description
      「研究成果報告書概要(和文)」より
  • [Book] バイオとナノの融合II-新生命科学の応用2007

    • Author(s)
      長堀, 紀子
    • Total Pages
      8
    • Publisher
      北海道大学出版会
    • Description
      「研究成果報告書概要(和文)」より
  • [Book] Methods in Enzymology2006

    • Author(s)
      Hinou, H.
    • Total Pages
      11
    • Publisher
      Academic Press
    • Description
      「研究成果報告書概要(和文)」より
  • [Book] Nanotechnology in Carbohydrate Chemistry2006

    • Author(s)
      Nagahori, N.
    • Total Pages
      17
    • Publisher
      Transworld Research Network
    • Description
      「研究成果報告書概要(和文)」より
  • [Patent(Industrial Property Rights)] 特許権2007

    • Inventor(s)
      西村, 紳一郎・比能, 洋
    • Industrial Property Rights Holder
      北海道大学
    • Industrial Property Number
      特願2007-133706
    • Filing Date
      2007-05-21
    • Description
      「研究成果報告書概要(和文)」より
  • [Patent(Industrial Property Rights)] 特許権2007

    • Inventor(s)
      西村, 紳一郎・比能, 洋
    • Industrial Property Rights Holder
      北海道大学
    • Industrial Property Number
      特願2007-158635
    • Filing Date
      2007-06-15
    • Description
      「研究成果報告書概要(和文)」より
  • [Patent(Industrial Property Rights)] 特許権2005

    • Inventor(s)
      西村, 紳一郎, 他3名
    • Industrial Property Rights Holder
      北海道大学他2社
    • Industrial Property Number
      PCT/JP2005/023162
    • Filing Date
      2005-12-16
    • Description
      「研究成果報告書概要(和文)」より
    • Overseas

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Published: 2010-02-04  

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