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High-speed and high-sensitive methods of carbohydrates based on mass isotopomer analysis with the aim of monitoring glycan dynamics

Research Project

Project/Area Number 24790301
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field General medical chemistry
Research InstitutionThe Institute of Physical and Chemical Research

Principal Investigator

NAKAJIMA Kazuki  独立行政法人理化学研究所, 脳科学総合研究センター, 研究員 (10442998)

Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywords糖鎖修飾 / 糖ヌクレオチド / 質量分析 / クロマトグラフィー / グルコース代謝 / シアル酸 / 糖鎖 / 糖タンパク質
Outline of Final Research Achievements

In order to clarify biological significance of glycan dynamics, we have developed a tracing method for UDP-GlcNAc synthesis and utilization, and GlcNAc utilization using 13C6-glucose and 13C2-glucosamine, respectively, followed by the analysis of mass isotopomers. Because nucleotide sugars an important building block in glycan biosynthesis. Analysis of isotopomers of UDP-HexNAc and CMP-NeuAc, labeled with 13C6-glucose, revealed the relative contributions of metabolic pathways leading to UDP-GlcNAc synthesis and utilization. Using 13C2-glucosamine, the diversity of glycan turnover was observed in each glycan. In pancreatic insulinoma cells, the labeling efficiency of CMP-NeuAc was lower compared with that in hepatoma cells, indicating that metabolic flows are responsible for the low sialylation in the insulinoma cells. Thus, our strategy should be useful for systematically tracing each stage of cellular glycan dynamics.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • Research Products

    (13 results)

All 2014 2013 2012

All Journal Article (7 results) (of which Acknowledgement Compliant: 1 results,  Peer Reviewed: 6 results) Presentation (6 results)

  • [Journal Article] Simultaneous Quantification of Nucleotide Sugar Metabolism by LC and LC-MS2014

    • Author(s)
      Kazuki Nakajima, Naoyuki Taniguchi
    • Journal Title

      Glycoscience: Biology and Medicine

      Volume: 1 Pages: 103-110

    • Related Report
      2014 Annual Research Report
    • Acknowledgement Compliant
  • [Journal Article] The absence of core fucose upregulates GnT-III and Wnt target genes: a possible mechanism for an adaptive response in terms of glycan function2014

    • Author(s)
      Kurimoto A, Kitazume S, Kizuka Y, Nakajima K, Oka R, Fujinawa R, Korekane H, Yamaguchi Y, Wada Y, Taniguchi N
    • Journal Title

      J Biol Chem. (Epub 2014 Mar10.)

      Volume: 289(17) Issue: 17 Pages: 11704-14

    • DOI

      10.1074/jbc.m113.502542

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Ceramide galactosyltransferase expression is regulated positively by Nkx2.2 and negatively by OLIG2.2014

    • Author(s)
      Okahara K, Kizuka Y, Kitazume S, Ota F, Nakajima K, Hirabayashi Y, Maekawa M, Yoshikawa T, Taniguchi N.
    • Journal Title

      Glycobiology

      Volume: 10 Issue: 10 Pages: 926-34

    • DOI

      10.1093/glycob/cwu042

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The Absence of Core Fucose Up-regulates GnT-III and Wnt Target Genes: A POSSIBLE MECHANISM FOR AN ADAPTIVE RESPONSE IN TERMS OF GLYCAN FUNCTION.2014

    • Author(s)
      Kurimoto A, Kitazume S, Kizuka Y, Nakajima K, Oka R, Fujinawa R, Korekane H, Yamaguchi Y, Wada Y, Taniguchi N
    • Journal Title

      J Biol Chem

      Volume: 289 Pages: 11704-11714

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Mass isotopomer analysis of metabolically labeled nucleotide sugars and N- and O-glycans for tracing nucleotide sugar metabolisms.2013

    • Author(s)
      Nakajima, K., Ito, E., Ohtsubo, K., Shirato, K., Takamiya, R., Kitazume, S., Angata, T., Taniguchi, N.
    • Journal Title

      Mol. & Cell Proteomics.

      Volume: 12 Pages: 2468-2080

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Metabolically programmed quality control system for dolichol-linked oligosaccharides.2013

    • Author(s)
      Harada, Y., Nakajima, K., Masahara-Negishi, Y., Freeze, H.H., Angata, T., Taniguchi, N., Suzuki, T.
    • Journal Title

      Proc. Natl. Acad. Sci. U.S.A

      Volume: 110 Pages: 19366-19371

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Identification of ectonucleotide pyrophosphatase/phosphodiesterase 3 (ENPP3) as a regulator of N-acetylglucosaminyltransferase GnT-IX (GnT-Vb).2013

    • Author(s)
      Korekane, H., Park, J.Y, Matsumoto, A., Nakajima, K., Takamatsu, S., Ohtsubo, K., Miyamoto, Y., Hanashima, S., Kanekiyo, K., Kitazume, S., Yamaguchi, Y., Matsuo, I., Taniguchi, N.
    • Journal Title

      J. Biol. Chem.

      Volume: 288 Pages: 27912-27926

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] 双性イオイ性カラム(ZIC)-HILIC-ESI-MSによる脳内グルコース化分子の異性体定量2014

    • Author(s)
      8. 中嶋 和紀、秋山 央子、田中 香織、谷口 直之、平林 義雄
    • Organizer
      第33回日本糖質学会年会
    • Place of Presentation
      名古屋
    • Year and Date
      2014-08-10 – 2014-08-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] Mass isotopomer analysis of metabolically labeled nucleotide sugars and N- and O-glycans for tracing nucleotide sugar metabolisms.2013

    • Author(s)
      Nakajima, K., Ito, E., Ohtsubo, K., Shirato, K., Takamiya, R., Kitazume, S., Angata, T., Taniguchi, N.
    • Organizer
      HUPO 12th Annual World Congress
    • Place of Presentation
      yokohama
    • Related Report
      2013 Research-status Report
  • [Presentation] 質量同位体測定によるグルコサミンの代謝動態の追跡2013

    • Author(s)
      中嶋和紀、伊藤恵実、大坪和明、白土健、高宮里奈、北爪しのぶ、安形高志、谷口直之
    • Organizer
      第32回糖質学会年会
    • Place of Presentation
      大阪
    • Related Report
      2013 Research-status Report
  • [Presentation] 質量同位体測定によるグルコサミンの代謝動態の追跡2013

    • Author(s)
      中嶋和紀、伊藤恵実、大坪和明、白土健、高宮里奈、北爪しのぶ、安形高志、谷口直之
    • Organizer
      第86回日本生化学会年会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Presentation] Tracing Strategy of GlcNAc Cycle by Mass Isotopomers Analysis2012

    • Author(s)
      Nakajima, K., Ito, E., Ohtsubo, K., Shirato, K., Takamiya, R., Kitazume, S., Angata, T., and Taniguchi, N.
    • Organizer
      HUPO 2012
    • Place of Presentation
      Boston
    • Related Report
      2012 Research-status Report
  • [Presentation] 質量同位体測定に基づくN-アセチルグルコサミンサイクルの追跡戦略2012

    • Author(s)
      中嶋和紀、伊藤恵実、大坪和明、白土健、高宮里奈、北爪しのぶ、安形高志、谷口直之
    • Organizer
      第85回日本生化学会年会
    • Place of Presentation
      福岡
    • Related Report
      2012 Research-status Report

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Published: 2013-05-31   Modified: 2019-07-29  

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