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Creation of functional cushion protein for a highly sensitive bimolecular interaction detection system by rational molecular design

Research Project

Project/Area Number 15K06582
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionOkayama University

Principal Investigator

Imanaka Hiroyuki  岡山大学, 自然科学研究科, 助教 (10379711)

Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords生体分子 / 固定化 / 相互作用 / 蛋白質 / ペプチド / リガンド / 超好熱菌 / バイオ分子 / タンパク質
Outline of Final Research Achievements

The applications of combinations of surface affinity peptide-tag and cushioning scaffold protein, CutA1 from a hyperthermophilic archaeon, were examined in detail to construct functional ligand biomolecular immobilized surfaces. Through the evaluations of non-labelled biomolecular interaction detection after immobilizations of rationally designed various CutA1s by quartz crystal microbalance (QCM), the importance of precise control of ligand orientation with avoiding steric hindrance to construct highly sensitive interaction detection system was clarified. On the other hand, an original affinity peptide of gold nanoparticle (AuNP-tag) was screened and its applications for sensitive colorimetric interaction detection was demonstrated with CutA1 scaffold. In addition, possible VHH antibody insertion site was successfully found in CutA1 and thus expansion of molecular design tolerance of CutA1 was verified.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (22 results)

All 2018 2017 2016 2015

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (20 results) (of which Int'l Joint Research: 8 results,  Invited: 3 results) Book (1 results)

  • [Journal Article] The use of a proteinaceous "cushion" with a polystyrene-binding peptide tag to control the orientation and function of a target peptide adsorbed to a hydrophilic polystyrene surface.2016

    • Author(s)
      Imanaka, H., D. Yamadzumi, K. Yanagita, N. Ishida, K. Nakanishi and K. Imamura
    • Journal Title

      Biotechnol Progress

      Volume: 32(2) Issue: 2 Pages: 527-534

    • DOI

      10.1002/btpr.2232

    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Presentation] Design of Functional Molecular Recognition Element by Utilizing Structurally Robust Protein Scaffold2017

    • Author(s)
      H. Imanaka, K. Date, H. Matoba, N. Ishida and K. Imamura
    • Organizer
      Young Asian Biological Engineers’ Community 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] CutA1を足場とした機能性分子認識素子の開発2017

    • Author(s)
      今中 洋行,伊達 弘輝,石田 尚之,今村 維克
    • Organizer
      化学工学会第49回秋季大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] CutA1をリガンド足場として利用した生体分子間相互作用高感度検出のための分子デザイン2017

    • Author(s)
      伊達 弘輝, 石田 尚之,今村 維克,清瀬 紀彦,宮崎 誠生,今中 洋行
    • Organizer
      第69回日本生物工学会大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 超好熱菌由来タンパク質CutA1を足場とした機能性分子認識素子のデザイン2017

    • Author(s)
      今中 洋行,伊達 弘輝,沖 拓哉,石田 尚之,今村 維克
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] CutA1を用いた分子デザインによりタンパク質間相互作用検出を高感度化する方法2017

    • Author(s)
      今中 洋行,伊達 弘輝,石田 尚之,今村 維克
    • Organizer
      日本農芸化学会2017年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 弱い生体分子間相互作用をCutA1の分子デザインを通じて感度よく検出する2017

    • Author(s)
      今中 洋行,伊達 弘輝,的場 晴香,沖 拓哉,石田 尚之,今村 維克
    • Organizer
      化学工学会第82年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Utilization of Hyperstable CutA1 as a Scaffold for Multiple Ligand Display in Biomolecular Recognition System2017

    • Author(s)
      H. Imanaka
    • Organizer
      2nd Korea-Japan smart biodesign workshop
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Influence of Orientation and Spatial Arrangement of Ligand Biomolecules on Functionality after Immobilization on Solid Materials2016

    • Author(s)
      Imanaka, H.*, K. Date, H. Matoba, and K. Imamura
    • Organizer
      Grobal Research Efforts on Energy and Nanomaterials (GREEN 2016)
    • Place of Presentation
      台北(台湾)
    • Year and Date
      2016-12-22
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] クッションタンパク質の分子デザインによるバイオ分子間相互作用の高感度化2016

    • Author(s)
      今中洋行,伊達弘輝,的場晴香,石田尚之,今村維克
    • Organizer
      真空・表面科学合同講演会:第36回表面科学学術講演会・第57回真空
    • Place of Presentation
      名古屋
    • Year and Date
      2016-11-29
    • Related Report
      2016 Research-status Report
  • [Presentation] Investigations of Biomolecular Interaction Detection System utilizing Proteinaceous Cushion2016

    • Author(s)
      Imanaka H.*, K. Date, H. Matoba, N. Ishida and K. Imamura
    • Organizer
      Young Asian Bioclogical Engineers’ Community 2016
    • Place of Presentation
      宮崎
    • Year and Date
      2016-10-27
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] CutA1及びVHH抗体を利用した高感度バイオセンシングシステムのデザイン2016

    • Author(s)
      伊達 弘輝, 石田 尚之, 今村 維克, 今中 洋行
    • Organizer
      第68回日本生物工学会大会
    • Place of Presentation
      富山
    • Year and Date
      2016-09-28
    • Related Report
      2016 Research-status Report
  • [Presentation] Molecular Design of Hyperthermostable Proteinaceous Cushion for Sensitive Biomolecular Interaction Detection System2016

    • Author(s)
      Imanaka H.*, K. Date, Y. Inaba, N. Ishida and K. Imamura
    • Organizer
      11th Interanational Congress on Extremophiles
    • Place of Presentation
      京都
    • Year and Date
      2016-09-12
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] リガンドおよびドメインの挿入がクッションタンパク質CutA1の構造安定性と相互作用検出感度に及ぼす影響2016

    • Author(s)
      伊達弘輝,稲葉優樹,石田尚之,今村維克,今中洋行
    • Organizer
      化学工学会第48回秋季大会
    • Place of Presentation
      徳島
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] 固体表面上に生体分子を機能的に固定化し,感度よく相互作用をみる方法2016

    • Author(s)
      今中洋行,伊達弘輝,的場晴香,石田尚之,今村維克
    • Organizer
      粉体工学会第52回夏期シンポジウム
    • Place of Presentation
      神戸
    • Year and Date
      2016-08-09
    • Related Report
      2016 Research-status Report
  • [Presentation] Molecular Design of Proteinaceous Cushion for Sensitive Biomolecular Interaction Detection System2016

    • Author(s)
      Imanaka H.*, K. Date, Y. Maehara, N. Ishida and K. Imamura
    • Organizer
      17th European Congress on Biotechnology
    • Place of Presentation
      クラクフ(ポーランド)
    • Year and Date
      2016-06-03
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] CutA1を基盤としたリガンドデザインによるバイオ分子間相互作用検出の高感度化2016

    • Author(s)
      今中 洋行,伊達 弘輝,前原 康秀,稲葉 優樹,石田 尚之,今村 維克
    • Organizer
      化学工学会 第81年会
    • Place of Presentation
      大阪
    • Year and Date
      2016-03-14
    • Related Report
      2015 Research-status Report
  • [Presentation] Design of Sensitive Biomolecular Interaction Detection System on Solid Surfaces2015

    • Author(s)
      Hiroyuki Imanaka, Toshinobu Kunikata, Miki Ishimaru, Runa Matsushita, Naoyuki Ishida, and Koreyoshi Imamura
    • Organizer
      Asian Congress on Biotechnology (ACB2015)
    • Place of Presentation
      Kuala Lumpur, Malaysia
    • Year and Date
      2015-11-18
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] 金ナノ粒子の機能化に資するアンカーペプチドの探索2015

    • Author(s)
      今中 洋行,重森 陽士郎,本多 裕之
    • Organizer
      第67回日本生物工学会
    • Place of Presentation
      鹿児島
    • Year and Date
      2015-10-27
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] Design of sensitive peptide-protein interaction detection system on solid surfaces2015

    • Author(s)
      Hiroyuki Imanaka
    • Organizer
      30th Anniversary Meeting and International Symposium of KSBB
    • Place of Presentation
      Incheon, Korea
    • Year and Date
      2015-10-14
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] 超好熱菌由来CutA1を基盤としたリガンドデザインと分子間相互作用検出2015

    • Author(s)
      今中 洋行,稲葉 優樹,松下 瑠奈,重森 陽士郎,石田 尚之,今村 維克
    • Organizer
      第15回日本蛋白質科学会
    • Place of Presentation
      徳島
    • Year and Date
      2015-06-25
    • Related Report
      2015 Research-status Report
  • [Book] 細胞・生体分子の固定化と機能発現2018

    • Author(s)
      今中洋行(黒田章夫監修)
    • Total Pages
      294
    • Publisher
      シーエムシー出版
    • ISBN
      9784781313269
    • Related Report
      2017 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2019-03-29  

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