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Development of a novel screening system for PPI-targeted drugs using cutinase fusion strategy

Research Project

Project/Area Number 24651260
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field Chemical biology
Research InstitutionOsaka University

Principal Investigator

TAKAGI Junichi  大阪大学, たんぱく質研究所, 教授 (90212000)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywords化合物スクリーニング / cutinase / ケミカルバイオロジー / セマフォリン / プレキシン / Wntシグナル伝達系 / ニカストリン
Research Abstract

A wide variety of methods have been developed to detect, analyze, and quantify protein interactions, including SPR, ITC, co-precipitation, affinity chromatography, ultracentrifugation, NMR, mass spectrometry, and ELISAs in vitro. While these methods are widely used to identify protein function, they quickly reach their limits when it comes to addressing the intricate, dynamic mechanics of a biological pathway. In contrast, cell-based PPIs assays are appropriate for wider applications. However, these assays have different limitations such as the low-contrast imaging, using fixed-cells, high-cost and a lot of step experiments. In this study, we have established the PPI assay system which observes the site-specifically labeled target proteins with a binding partner under physiological context in live cells.

Report

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

    (37 results)

All 2014 2013 2012 Other

All Journal Article (9 results) (of which Peer Reviewed: 7 results) Presentation (26 results) (of which Invited: 10 results) Book (2 results)

  • [Journal Article] PA tag: a versatile protein tagging system using a super high affinity antibody against a dodecapeptide derived from human podoplanin2014

    • Author(s)
      Fujii Y, Kaneko M, Neyazaki M, Nogi T, Kato Y, Takagi J.
    • Journal Title

      Protein Expr Purif

      Volume: 95 Pages: 240-247

    • DOI

      10.1016/j.pep.2014.01.009

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] A multi-purpose fusion tag derived from an unstructured and hyper-acidic region of the amyloid precursor protein.2013

    • Author(s)
      Sangawa, T., Tabata, S., Suzuki, K., Saeki, Y., Tanaka, K., and Takagi, J.
    • Journal Title

      Protein Science

      Volume: 22 Issue: 6 Pages: 840-850

    • DOI

      10.1002/pro.2254

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] セマフォリン関連分子群の立体構造2013

    • Author(s)
      松永幸子、高木淳一
    • Journal Title

      実験医学

      Volume: 31 Pages: 523-530

    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Journal Article] A point mutation in Semaphorin 4A associates with defective endosomal sorting and causes retinal degeneration2013

    • Author(s)
      Nojima S, et al.
    • Journal Title

      Nat Commun

      Volume: 4 Issue: 1 Pages: 1406-1406

    • DOI

      10.1038/ncomms2420

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp12012

    • Author(s)
      Kato K, Nishimasu H, Mihara E, Ishitani R, Takagi J, Aoki J, Nureki O
    • Journal Title

      Acta Crystallogr

      Volume: F68 Issue: 7 Pages: 778-782

    • DOI

      10.1107/s1744309112019306

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Higher-order architecture of cell adhesion mediated by polymorphic synaptic adhesion molecules neurexin and neuroligin2012

    • Author(s)
      Tanaka, H., Miyazaki, N., Matoba, K., Nogi, T., Iwasaki, K., and Takagi, J
    • Journal Title

      Cell Reports

      Volume: 2(1) Issue: 1 Pages: 101-110

    • DOI

      10.1016/j.celrep.2012.06.009

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Schwann cell myelination requires integration of laminin activities2012

    • Author(s)
      Karen K. McKee
    • Journal Title

      Journal of Cell Science

      Volume: 125 Pages: 4609-4619

    • DOI

      10.1242/jcs.107995

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of Enpp1, an extracellular glycoprotein involved in bone mineralization and insulin signaling2012

    • Author(s)
      Kato K, Nishimasu H, Okudaira S, Mihara E, Ishitani R, Takagi J, Aoki J, Nureki O
    • Journal Title

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

      Volume: 109 Issue: 42 Pages: 16876-16881

    • DOI

      10.1073/pnas.1208017109

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] LDL受容体ファミリーの構造と蛋白質間相互作用2012

    • Author(s)
      北郷悠、高木淳一
    • Journal Title

      The Lipid

      Volume: 23 Pages: 56-62

    • Related Report
      2013 Final Research Report 2012 Research-status Report
  • [Presentation] 創薬ターゲットの高品質生産を可能にする技術革新2014

    • Author(s)
      高木淳一
    • Organizer
      大阪大学未来戦略機構シンポジウム「アカデミア基盤研究から未来創薬へ」
    • Place of Presentation
      東京学士会館(東京)
    • Year and Date
      2014-03-27
    • Related Report
      2013 Final Research Report
  • [Presentation] Boosting the protein production pipeline by the use ofcustom-made epitope tag system2014

    • Author(s)
      Takagi J
    • Organizer
      IPR Seminar"Antibody Design, Modeling and Applications"
    • Place of Presentation
      IPR, Osaka University(大阪)
    • Year and Date
      2014-01-15
    • Related Report
      2013 Final Research Report
  • [Presentation] 創薬関連分子の構造生物学の最前線2013

    • Author(s)
      高木淳一
    • Organizer
      千里ライフサイエンスセミナー
    • Place of Presentation
      千里ライフサイエンスセンター(大阪)
    • Year and Date
      2013-10-16
    • Related Report
      2013 Final Research Report
  • [Presentation] 神経ガイダンス因子セマフォリン3AのC末端におけるニューロピリン受容体との結合モチーフの同定2013

    • Author(s)
      松永幸子
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取)
    • Year and Date
      2013-06-14
    • Related Report
      2013 Final Research Report
  • [Presentation] PAタグを用いた新規アフィニティータグシステムの確立2013

    • Author(s)
      藤井勇樹
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取)
    • Year and Date
      2013-06-13
    • Related Report
      2013 Final Research Report
  • [Presentation] Multi-faceted approach to analyze structure and function of integrins2013

    • Author(s)
      Takagi J
    • Organizer
      Gordon Research Conference (Fibronectin, Integrins, & Related molecules)
    • Place of Presentation
      Ventura Beach Marriott(USA)
    • Year and Date
      2013-02-14
    • Related Report
      2013 Final Research Report
  • [Presentation] Revisiting the NRP1-Binding Determinant in the C-Terminal Region of SEMA3A2013

    • Author(s)
      Matsunaga Y
    • Organizer
      EMBO Workshop "Semaphorin function and mechanism of action"
    • Place of Presentation
      Abbaye des Vaux de Cernay (France)
    • Related Report
      2013 Final Research Report
  • [Presentation] REVISITING THE NRP1-BINDING DETERMINANT IN THE C-TERMINAL REGION OF SEMA3A2013

    • Author(s)
      松永 幸子
    • Organizer
      EMBO Workshop "Semaphorin function and mechanism of action"
    • Place of Presentation
      Abbaye des Vaux de Cernay, France
    • Related Report
      2013 Annual Research Report
  • [Presentation] Higher-order Architecture of Cell Adhesion Mediated by Polymorphic Synaptic Adhesion Molecules Neurexin and Neuroligin2012

    • Author(s)
      Takagi J
    • Organizer
      第35回分子生物学会大会
    • Place of Presentation
      福岡国際会議場(福岡
    • Year and Date
      2012-12-13
    • Related Report
      2013 Final Research Report
  • [Presentation] 免疫学研究における構造生物学のインパクト2012

    • Author(s)
      高木淳一
    • Organizer
      第41回日本免疫学会学術集会
    • Place of Presentation
      神戸ポートアイランド(兵庫)
    • Year and Date
      2012-12-07
    • Related Report
      2013 Final Research Report
  • [Presentation] "構造神経科学のすすめ"−立体構造情報を使いこなすクールなニューロサイエンティストになろう−2012

    • Author(s)
      高木淳一
    • Organizer
      第35回日本神経科学会大会
    • Place of Presentation
      名古屋国際会議場(愛知)
    • Year and Date
      2012-09-21
    • Related Report
      2013 Final Research Report
  • [Presentation] Correlation technology to fill the gap between X-ray crystallography and microscopic imaging2012

    • Author(s)
      Takagi J
    • Organizer
      International Integrin Signaling Meeting
    • Place of Presentation
      Universitatszentrum Obergurgl, Tirol (Austria)
    • Year and Date
      2012-09-16
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛋白質の立体構造から理解する細胞外シグナル授受のしくみ2012

    • Author(s)
      高木淳一
    • Organizer
      第12回日本抗加齢医学会
    • Place of Presentation
      パシフィコ横浜(神奈川)
    • Year and Date
      2012-06-24
    • Related Report
      2013 Final Research Report
  • [Presentation] PPI阻害物質探索のためのprotein-based screening2012

    • Author(s)
      高木淳一
    • Organizer
      第12回日本蛋白質科学会年会
    • Place of Presentation
      名古屋国際会議場(愛知)
    • Year and Date
      2012-06-22
    • Related Report
      2013 Final Research Report
  • [Presentation] 創薬関連分子の構造生物学の最前線

    • Author(s)
      高木 淳一
    • Organizer
      千里ライフサイエンスセミナー
    • Place of Presentation
      千里ライフサイエンスセンター(大阪府)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Boosting the protein production pipeline by the use ofcustom-made epitope tag system

    • Author(s)
      Junichi Takagi
    • Organizer
      IPR Seminar “Antibody Design, Modeling and Applications”
    • Place of Presentation
      IPR, Osaka Univ.(大阪府)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 創薬ターゲットの高品質生産を可能にする技術革新

    • Author(s)
      高木 淳一
    • Organizer
      大阪大学未来戦略機構創薬シンポジウム「アカデミア基盤研究から未来創薬へ」
    • Place of Presentation
      東京 学士会館(東京都)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 神経ガイダンス因子セマフォリン3AのC末端におけるニューロピリン受容体との結合モチーフの同定

    • Author(s)
      松永 幸子
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] PAタグを用いた新規アフィニティータグシステムの確立

    • Author(s)
      藤井 勇樹
    • Organizer
      第13回日本蛋白質科学会年会
    • Place of Presentation
      とりぎん文化会館(鳥取県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] PPI阻害物質探索のためのprotein-based screening

    • Author(s)
      高木淳一
    • Organizer
      第12回日本蛋白質科学会年会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] 蛋白質の立体構造から理解する細胞外シグナル授受のしくみ

    • Author(s)
      高木淳一
    • Organizer
      第12回日本抗加齢医学会
    • Place of Presentation
      パシフィコ横浜
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] Correlation technology to fill the gap between X-ray crystallography and microscopic imaging

    • Author(s)
      Junichi Takagi
    • Organizer
      "International Integrin Signaling Meeting"
    • Place of Presentation
      Universitaetszentrum Obergurgl, Tirol - Austria
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] “構造神経科学”のすすめ-立体構造情報を使いこなすクールなニューロサイエンティストになろう-

    • Author(s)
      高木淳一
    • Organizer
      第35回日本神経科学会大会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] 免疫学研究における構造生物学のインパクト

    • Author(s)
      高木淳一
    • Organizer
      第41回日本免疫学会学術集会
    • Place of Presentation
      神戸ポートアイランド
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] Higher-order Architecture of Cell Adhesion Mediated by Polymorphic Synaptic Adhesion Molecules Neurexin and Neuroligin

    • Author(s)
      Junichi Takagi
    • Organizer
      第35回日本分子生物学会大会
    • Place of Presentation
      福岡国際会議場
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] Multi-faceted approach to analyze structure and function of integrins

    • Author(s)
      Junichi Takagi
    • Organizer
      Gordon Research Conference (Fibronectin, Integrins, & Related molecules)
    • Place of Presentation
      Ventura Beach Marriott, USA
    • Related Report
      2012 Research-status Report
    • Invited
  • [Book] 動物細胞を用いる膜タンパク質生産2013

    • Author(s)
      高木淳一
    • Publisher
      化学同人
    • Related Report
      2013 Final Research Report
  • [Book] 動物細胞を用いる膜タンパク質生産2013

    • Author(s)
      高木 淳一
    • Publisher
      「膜タンパク質構造研究」(岩田 想 編)化学同人
    • Related Report
      2013 Annual Research Report

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

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