• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Reconstitution of private chaperone systems using synthetic binding proteins

Research Project

Project/Area Number 15H05370
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Biomolecular chemistry
Research InstitutionOkayama University

Principal Investigator

Yasui Norihisa  岡山大学, 医歯薬学総合研究科, 准教授 (90467514)

Project Period (FY) 2015-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥23,660,000 (Direct Cost: ¥18,200,000、Indirect Cost: ¥5,460,000)
Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2016: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2015: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Keywords人工タンパク質 / タンパク質 / 進化分子工学 / 合成生物学 / 分子シャペロン / 一本鎖モネリン / 人工結合タンパク質 / 単鎖モネリン / 蛋白質
Outline of Final Research Achievements

In this study, to clarify the correlation between the fold-specific substrate recognition and the function of the private chaperones, the system for generation of the synthetic binding proteins that specifically bind to the target proteins has been developed. The phage display libraries of the synthetic binding proteins using single-chain monellin as a non-antibody molecular scaffold were designed and constructed. By sorting the phage display libraries against the green fluorescent protein variant (GFPuv), the synthetic binding protein that has the ability to bind to GFPuv (GBP-1) was generated. The characteristics of GBP-1 revealed in this study indicate that single-chain monellin is useful as a non-antibody molecule backbone of the synthetic binding proteins.

Academic Significance and Societal Importance of the Research Achievements

本研究では,ファージディスプレイライブラリーの作製とその選別により検討を重ねた結果,一本鎖モネリンを非抗体分子骨格として利用すると,標的分子に親和性を示す人工結合タンパク質を作製できることを示した。本研究で構築した一本鎖モネリンを非抗体分子骨格とする人工結合タンパク質ライブラリーは,プライベートシャペロンの機能解析に留まらず,様々な研究で利用することができる点でも意義がある。

Report

(5 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (18 results)

All 2019 2018 2017 2016 2015

All Journal Article (8 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 5 results,  Open Access: 5 results) Presentation (10 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Specific modification at the C-terminal lysine residue of the green fluorescent protein variant, GFPuv, expressed in Escherichia coli2019

    • Author(s)
      Takahiro Nakatani, Norihisa Yasui, Issei Tamura & Atsuko Yamashita
    • Journal Title

      Scientific Reports

      Volume: 9 Issue: 1 Pages: 4722-4722

    • DOI

      10.1038/s41598-019-41309-8

    • NAID

      120006777526

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Functional analyses of specific protein-protein interaction networks using affinity clamping technology2017

    • Author(s)
      安井 典久
    • Journal Title

      生化学

      Volume: 89 Issue: 2 Pages: 293-297

    • DOI

      10.14952/SEIKAGAKU.2017.890293

    • NAID

      40021203455

    • ISSN
      0037-1017
    • Year and Date
      2017-04-25
    • Related Report
      2017 Annual Research Report
  • [Journal Article] Structural basis for perception of diverse chemical substances by T1r taste receptors2017

    • Author(s)
      Nuemket Nipawan、Yasui Norihisa、Kusakabe Yuko、Nomura Yukiyo、Atsumi Nanako、Akiyama Shuji、Nango Eriko、Kato Yukinari、Kaneko Mika K.、Takagi Junichi、Hosotani Maiko、Yamashita Atsuko
    • Journal Title

      Nat Commun.

      Volume: 8 Issue: 1 Pages: 15530-15530

    • DOI

      10.1038/ncomms15530

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Structural basis for ligand capture and release by the endocytic receptor ApoER2.2017

    • Author(s)
      Hirai H, Yasui N, Yamashita K, Tabata S, Yamamoto M, Takagi J, Nogi T
    • Journal Title

      EMBO reports

      Volume: 18 Issue: 6 Pages: 982-999

    • DOI

      10.15252/embr.201643521

    • NAID

      130007420865

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Immunoaffinity Purification of the Glycosylated Extracellular Fragment of Mouse Plexin A2 Produced in a Mammalian Expression System.2017

    • Author(s)
      Nogi T, Mihara E, Yasui N, Takagi J
    • Journal Title

      Methods in Molecular Biology

      Volume: 1493 Pages: 57-72

    • DOI

      10.1007/978-1-4939-6448-2_4

    • ISBN
      9781493964468, 9781493964482
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Taste substance binding elicits conformational change of taste receptor T1r heterodimer extracellular domains2016

    • Author(s)
      E. Nango, S. Akiyama, S. Maki-Yonekura, Y. Ashikawa, Y. Kusakabe, E. Krayukhina, T. Maruno, S. Uchiyama, N. Nuemket, K. Yonekura, M. Shimizu, N. Atsumi, N. Yasui, T. Hikima, M. Yamamoto, Y. Kobayashi, A. Yamashita
    • Journal Title

      Sci. Rep

      Volume: 6 Issue: 1 Pages: 25745-25745

    • DOI

      10.1038/srep25745

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 翻訳後修飾を含むアミノ酸配列を高特異的に認識する人工結合タンパク質2015

    • Author(s)
      安井 典久, 服部 峰充, 小出 昌平
    • Journal Title

      バイオサイエンスとインダストリー

      Volume: 73 Pages: 292-296

    • NAID

      40020536333

    • Related Report
      2015 Annual Research Report
  • [Journal Article] 人工タンパク質を用いたタンパク質間相互作用構築による細胞内シグナル伝達ネットワークの詳細解析2015

    • Author(s)
      安井 典久
    • Journal Title

      生物物理

      Volume: 55 Pages: 151-153

    • NAID

      130005072027

    • Related Report
      2015 Annual Research Report
    • Open Access
  • [Presentation] 一本鎖モネリンを分子骨格とする人工結合タンパク質ファージディスプレイライブラリーの構築2018

    • Author(s)
      安井 典久,山下 敦子
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 分子骨格として一本鎖モネリンを用いたGFPを標的とする人工結合タンパク質の作製とその分子特性解析2018

    • Author(s)
      安井 典久,山下 敦子
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] サーマルシフトアッセイによる味覚受容体細胞外領域に対するアミノ酸の結合解析2018

    • Author(s)
      吉田 高志,安井 典久,渥美 菜奈子, 山下 敦子
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 表面プラズモン共鳴法による味覚受容体T1R細胞内C末端領域とカルシウム結合タンパク質の相互作用の速度論的解析2018

    • Author(s)
      吉田 涼希,安井 典久,山下 敦子
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] キチン結合ドメインを分子骨格とする人工結合タンパク質ファージディスプレイライブラリーの作製2017

    • Author(s)
      安井 典久
    • Organizer
      ConBio2017
    • Related Report
      2017 Annual Research Report
  • [Presentation] X線結晶構造解析によるメダカT1r2a/T1r3細胞外領域への無機金属イオン結合解析2017

    • Author(s)
      渥美 菜奈子, 安井 典久, 山下 敦子
    • Organizer
      日本味と匂学会第51回大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Expression, purification, and characterization of the entire heterodimeric extracellular regions of fish taste receptor2017

    • Author(s)
      Hiroki Maruhashi, Daisuke Noshiro, Norihisa Yasui, Toshio Ando, Atsuko Yamashita
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Fish taste receptor type 1 (T1R) extracellular domains exhibit broad specificity for ligand binding2016

    • Author(s)
      Nanako Atsumi, Yukiyo Nomura, Maude W. Baldwin, Norihisa Yasui, AtsukoYamashita
    • Organizer
      The 15th International Symposium on Molecular and Neural Mechanisms of Taste and Olfactory Perception
    • Place of Presentation
      Fukuoka, Japan
    • Year and Date
      2016-12-04
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 大腸菌組換え発現したGFPで見いだされるC末端Lys残基の翻訳後修飾2016

    • Author(s)
      安井 典久, 中谷 隆寛, 山下 敦子
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-12-01
    • Related Report
      2016 Annual Research Report
  • [Presentation] 味覚受容体T1R細胞内領域とCa2+結合タンパク質の相互作用に関する研究2016

    • Author(s)
      伊藤 愛優美, 安井 典久,Nipawan Nuemket, 山下 敦子
    • Organizer
      日本薬学会第136回年会
    • Place of Presentation
      横浜
    • Year and Date
      2016-03-29
    • Related Report
      2015 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2020-03-30  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi