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Development and demonstration of novel purification techniques for disease-specific superficial proteins

Research Project

Project/Area Number 16K14680
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

Inoue Tsuyoshi  大阪大学, 工学研究科, 教授 (20263204)

Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsプロテオミクス解析 / 質量分析 / 表在性蛋白質 / 抗原探索 / 非天然ビオチン化ラベル / 改変型ストレプトアビジン / CTC / 血管内皮 / プロテオミクス / ラベル化技術 / 蛋白質精製 / 人工ビオチン / 抗体開発 / 診断薬開発 / 抗体医薬 / イメージング / 診断薬
Outline of Final Research Achievements

Regarding the labeling reagent, succeeded in synthesizing a derivative in which a succinimide group (NHS group) was added to the non-natural biotin derivatives.
As a result of experiments and proteomics analyses of vascular endothelium using mouse, it was found that the surface protein were only purified and the endogenous biotinylated proteins could be excluded by using the new derivatives. Proteomic analysis was also performed on diseased mice seeded with A20 lymphoma. Analyses of the vascular endothelial proteins that increased by 1.5 times or more in tumor-bearing mice were performed. There are 147 kinds of proteins listed only by the conventional method, and 28 kinds of proteins commonly listed in the conventional method and the novel method. However, it turned out that there were 53 kinds of proteins that were only found to be enhanced by the novel method. Validation is now under study.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (5 results)

All 2018 2017

All Journal Article (4 results) (of which Peer Reviewed: 4 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Patent(Industrial Property Rights) (1 results) (of which Overseas: 1 results)

  • [Journal Article] Crystal structure of secretory abundant heat soluble protein 4 from one of the toughest “water bears” micro-animals Ramazzottius varieornatus2018

    • Author(s)
      Fukuda Yohta、Inoue Tsuyoshi
    • Journal Title

      Protein Science

      Volume: 印刷中 Issue: 5 Pages: 993-999

    • DOI

      10.1002/pro.3393

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] New molecular packing in a crystal of pseudoazurin from Alcaligenes faecalis: a double-helical arrangement of blue copper.2017

    • Author(s)
      Yohta Fukuda, Eiichi Mizohata and Tsuyoshi Inoue
    • Journal Title

      Acta Cryst. F

      Volume: 73 Issue: 3 Pages: 159-166

    • DOI

      10.1107/s2053230x17002631

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Structural insights into a secretory abundant heat-soluble protein from an anhydrobiotic tardigrade, Ramazzottius?varieornatus2017

    • Author(s)
      Fukuda Yohta、Miura Yoshimasa、Mizohata Eiichi、Inoue Tsuyoshi
    • Journal Title

      FEBS letters

      Volume: 591 Issue: 16 Pages: 2458-2469

    • DOI

      10.1002/1873-3468.12752

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Active site geometry of a novel aminopropyltransferase for biosynthesis of hyperthermophile-specific branched-chain polyamine2017

    • Author(s)
      R. Hidese, K. M. Tse, S. Kimura, E. Mizohata, J. Fujita, Y. Horai, N. Umezawa, T. Higuchi, M. Niitsu, T. Oshima, T. Imanaka, T. Inoue, and S. Fujiwara
    • Journal Title

      The FEBS Journal

      Volume: 284 Issue: 21 Pages: 3684-3701

    • DOI

      10.1111/febs.14262

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Patent(Industrial Property Rights)] 抗体医薬の開発に資する創薬標的タンパク質の同定方法及び標的タンパク質に対する抗体の製造方法2018

    • Inventor(s)
      鈴木常司, 戸谷由之, 番場伸一, 鎌田春彦, 井上豪, 他
    • Industrial Property Rights Holder
      鈴木常司, 戸谷由之, 番場伸一, 鎌田春彦, 井上豪, 他
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Related Report
      2017 Annual Research Report
    • Overseas

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Published: 2016-04-21   Modified: 2019-03-29  

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