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Sequential enzyme complex formation on DNA

Research Project

Project/Area Number 26630419
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Biofunction/Bioprocess
Research InstitutionThe University of Tokyo

Principal Investigator

Hirakawa Hidehiko  東京大学, 工学(系)研究科(研究院), 助教 (90451799)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords酵素複合体 / 核内増殖抗原 / 古細菌 / Metallosphaera sedula / DNA / リガーゼ
Outline of Final Research Achievements

We tried to develop a method that arrange the order of multiple enzymes on DNA to construct artificial multienzyme complexes that catalyze sophisticated sequential or concerted reactions. To retain enzymes on DNA, we focused on a DNA sliding clamp. A stable DNA sliding clamp was discovered from a archaeon and then characterized. A DNA-binding protein was immobilized on DNA at the starting point of complex formation to avoid drop-off of proteins fused to the DNA sliding clamp. As as result, we successfully constructed multienzyme complexes on DNA.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (9 results)

All 2016 2015 2014

All Presentation (9 results) (of which Int'l Joint Research: 3 results)

  • [Presentation] 配列特異的なDNA結合タンパク質を利用したシトクロムP450システムの再構成2016

    • Author(s)
      畠山洋平、平川秀彦、長棟輝行
    • Organizer
      化学工学会第81年会
    • Place of Presentation
      関西大学(大阪府、吹田市)
    • Year and Date
      2016-03-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] DNA結合棚パク質を介したシトクロムP450システムの再構成2015

    • Author(s)
      畠山洋平、平川秀彦、長棟輝行
    • Organizer
      第67回日本生物工学会大会
    • Place of Presentation
      城山観光ホテル(鹿児島、鹿児島市)
    • Year and Date
      2015-10-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] PCNAヘテロ三量化を利用したDNA上への多酵素集積2015

    • Author(s)
      岩田史也、平川秀彦、長棟輝行
    • Organizer
      第67回日本生物工学会大会
    • Place of Presentation
      城山観光ホテル(鹿児島、鹿児島市)
    • Year and Date
      2015-10-28
    • Related Report
      2015 Annual Research Report
  • [Presentation] PCNAヘテロ三量化を利用したDNA上への多酵素集積2015

    • Author(s)
      岩田史也、平川秀彦、長棟輝行
    • Organizer
      酵素工学研究会 第74回講演会
    • Place of Presentation
      東京大学(東京都、文京区)
    • Year and Date
      2015-10-16
    • Related Report
      2015 Annual Research Report
  • [Presentation] Reconstitution of a Bacterial Cytochrome P450 System on DNA Scaffold2015

    • Author(s)
      Yohei Hatakeyama, Hidehiko Hirakawa, Teruyuki Nagamune
    • Organizer
      RIKEN Symposium “Metals in Biology” in Wako
    • Place of Presentation
      理化学研究所(埼玉県、和光市)
    • Year and Date
      2015-06-17
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Reconstitution of cytochrome P450 system utlizing DNA binding protein2015

    • Author(s)
      Yohei Hatakeyama, Hidehiko Hirakawa, Teruyuki Nagamune
    • Organizer
      19th International Conference on Cytochrome P450
    • Place of Presentation
      国立オリンピック記念青少年総合センター(東京都、渋谷区)
    • Year and Date
      2015-06-13
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Stable multienzyme complex formation using PCNA from Metallosphaera sedula2015

    • Author(s)
      Fumiya Iwata, Hidehiko Hirakawa, Teruyuki Nagamune
    • Organizer
      19th International Conference on Cytochrome P450
    • Place of Presentation
      国立オリンピック記念青少年総合センター(東京都、渋谷区)
    • Year and Date
      2015-06-13
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] クレン古細菌由来核内増殖抗原のDNA結合性の評価2014

    • Author(s)
      岩田史也、平川秀彦、長棟輝行
    • Organizer
      第87回日本生化学会大会
    • Place of Presentation
      京都・京都国際会館
    • Year and Date
      2014-10-18
    • Related Report
      2014 Research-status Report
  • [Presentation] DNA結合タンパク質を利用した細菌由来シトクロムP450システムの再構成2014

    • Author(s)
      平川秀彦、畠山洋平、岩田史也、長棟輝行
    • Organizer
      第66回日本生物工学会大会
    • Place of Presentation
      札幌・札幌コンベンションセンター
    • Year and Date
      2014-09-09
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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