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Identification of reaction intermediates in iron-sulfur cluster biogenesis for understanding its reaction mechanism

Research Project

Project/Area Number 17K14510
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Bio-related chemistry
Research InstitutionSaitama University

Principal Investigator

FUJISHIRO Takashi  埼玉大学, 理工学研究科, 助教 (20740450)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords鉄硫黄クラスター / 生合成 / 反応中間体 / X線結晶構造解析 / PLP / Chelatase / 枯草菌 / 構造ゲノム / 硫黄 / システイン / 鉄 / 触媒 / 酵素
Outline of Final Research Achievements

Biosynthetic machineries (NIF, ISC, SUF, SUF-like machineries) for the iron-sulfur (Fe-S) clusters, which are biologically essential inorganic cofactors, were investigated by elucidating catalytic intermediates of sulfur-mobilizing enzymes. Two cysteine desulfurases, NifS in NIF machinery and SufS in SUF-like machinery were analyzed by X-ray crystallography. This resulted in elucidation of common intermediate PLP-L-cysteine and different conformational changes of their catalytic loops toward the sulfur-mobilization. Also, X-ray crystallographic analysis of SufS-SufU complex revealed a molecular mechanism in which the Zn-coordination structural change of SufU triggered the sulfur-mobilization from SufS to SufU.

Academic Significance and Societal Importance of the Research Achievements

これまで鉄硫黄クラスター生合成系酵素の生化学的解析、遺伝学的解析、立体構造解析は数多くなされてきた。しかしながら、酵素それぞれの反応中間体の解析に焦点を当てて、生合成機構を化学的な観点から詳細に調べた例は多くなく、そのため本研究は挑戦的な試みであった。鉄硫黄クラスターの無機硫黄供給系の化学的な理解により、4種の異なる生合成系の多様性や分子進化について、分子構造の観点から考察が可能となったため、今後、鉄硫黄クラスターが環境に合わせた生物の進化・多様化によりどのような特性を獲得したかについて、生物学的な知見を得ることができるようになると期待される。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (25 results)

All 2020 2019 2018 2017 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Open Access: 1 results) Presentation (17 results) (of which Int'l Joint Research: 4 results,  Invited: 3 results) Book (1 results) Remarks (1 results)

  • [Journal Article] Snapshots of PLP‐substrate and PLP‐product external aldimines as intermediates in two types of cysteine desulfurase enzymes2019

    • Author(s)
      Nakamura Ryosuke、Hikita Masahide、Ogawa Shoko、Takahashi Yasuhiro、Fujishiro Takashi
    • Journal Title

      The FEBS Journal

      Volume: 287 Issue: 6 Pages: 1138-1154

    • DOI

      10.1111/febs.15081

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure of sirohydrochlorin ferrochelatase SirB: the last of the structures of the class II chelatase family2019

    • Author(s)
      Fujishiro Takashi、Shimada Yukino、Nakamura Ryosuke、Ooi Miho
    • Journal Title

      Dalton Transactions

      Volume: 48 Issue: 18 Pages: 6083-6090

    • DOI

      10.1039/c8dt04727h

    • Related Report
      2019 Annual Research Report 2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Identification of IscU residues critical for de novo iron-sulfur cluster assembly2019

    • Author(s)
      Tanaka Naoyuki、Yuda Eiki、Fujishiro Takashi、Hirabayashi Kei、Wada Kei、Takahashi Yasuhiro
    • Journal Title

      Molecular Microbiology

      Volume: 112 Issue: 6 Pages: 1769-1783

    • DOI

      10.1111/mmi.14392

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Distinct roles for U-type proteins in iron-sulfur cluster biosynthesis revealed by genetic analysis of the Bacillus subtilis sufCDSUB operon2018

    • Author(s)
      Yokoyama Nao、Nonaka Chihiro、Ohashi Yukari、Shioda Masaharu、Terahata Takuya、Chen Wen、Sakamoto Kotomi、Maruyama Chihiro、Saito Takuya、Yuda Eiki、Tanaka Naoyuki、Fujishiro Takashi、Kuzuyama Tomohisa、Asai Kei、Takahashi Yasuhiro
    • Journal Title

      Mol Microbiol

      Volume: 107 Issue: 6 Pages: 688-703

    • DOI

      10.1111/mmi.13907

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Zinc-Ligand Swapping Mediated Complex Formation and Sulfur Transfer between SufS and SufU for Iron?Sulfur Cluster Biogenesis in Bacillus subtilis2017

    • Author(s)
      Fujishiro Takashi、Terahata Takuya、Kunichika Kouhei、Yokoyama Nao、Maruyama Chihiro、Asai Kei、Takahashi Yasuhiro
    • Journal Title

      J Am Chem Soc

      Volume: 139 Issue: 51 Pages: 18464-18467

    • DOI

      10.1021/jacs.7b11307

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Mapping the key residues of SufB and SufD essential for biosynthesis of iron-sulfur clusters2017

    • Author(s)
      Yuda Eiki、Tanaka Naoyuki、Fujishiro Takashi、Yokoyama Nao、Hirabayashi Kei、Fukuyama Keiichi、Wada Kei、Takahashi Yasuhiro
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 9387-9387

    • DOI

      10.1038/s41598-017-09846-2

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 鉄硫黄クラスター生合成系のPLP依存型酵素SufSのシクロセリンによる阻害機構の解明2020

    • Author(s)
      小川 翔子, 中村 亮裕, 小松 茉里佳, 引田 理英, 藤城 貴史, 高橋 康弘
    • Organizer
      日本化学会第100春季年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 鉄硫黄クラスター生合成SUF-likeマシナリーにおける亜鉛依存型硫黄運搬タンパク質SufUの構造と機能2019

    • Author(s)
      藤城 貴史, 寺畑 拓也, 島田 侑希乃, 高橋 康弘
    • Organizer
      第92回日本生化学大会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 特殊な環境に生育する微生物由来の鉄硫黄クラスター生合成系の試験管再構成の試み2019

    • Author(s)
      國近 航平、藤城 貴史、山川 誠、和田 啓、高橋 康弘
    • Organizer
      第92回日本生化学大会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 異なる2つのグループに属するシステインデスルフラーゼに対する阻害剤の作用機構の解析2019

    • Author(s)
      中村 亮裕、小松 茉莉佳、小川 翔子、國近 航平、藤城 貴史、高橋 康弘
    • Organizer
      第92回日本生化学大会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 鉄硫黄クラスター生合成系における硫黄供給酵素のシステイン脱硫反応機構2019

    • Author(s)
      中村 亮裕、藤城 貴史、高橋 康弘
    • Organizer
      2019年度 日本生化学会関東支部例会-ノーベル賞と生化学-
    • Related Report
      2019 Annual Research Report
  • [Presentation] Structure-function relationship of sirohydrochlorin ferrochelatase SirB2019

    • Author(s)
      T. Fujishiro, Y. Shimada, R. Nakamura, M. Ooi
    • Organizer
      19th International Conference on Biological Inorganic Chemistry (ICBIC19)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] X-ray crystallographic snapshots of catalytic intermediates of cysteine desulfurases2019

    • Author(s)
      R. Nakamura, T. Fujishiro, Y. Takahashi
    • Organizer
      19th International Conference on Biological Inorganic Chemistry (ICBIC19)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] X-ray crystallographic snapshots of sulfur mobilization to iron-sulfur cluster biosynthesis2018

    • Author(s)
      Takashi Fujishiro
    • Organizer
      The International Symposium on Bioinorganic Chemistry 201
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] [Fe]-Hydrogenase-cofactor biosynthesis2018

    • Author(s)
      Takashi Fujishiro
    • Organizer
      The 43rd International Conference on Coordination Chemistry (ICCC2018)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Exploring an unusual Cys-rich motif-containing Fe-S enzyme2018

    • Author(s)
      Takashi Fujishiro, Miho Ooi, Yasuhiro Takahashi
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] Zinc-mediated sulfur acquisition by SufS-SufU complex involved in biosynthesis of iron-sulfur cluster in Bacillus subtilis2018

    • Author(s)
      Takashi Fujishiro, Takuya Terahata, Kunichika Kouhei, Chihiro Maruyama, Kei Asai, Yasuhiro Takahashi
    • Organizer
      錯体化学会第68回討論会
    • Related Report
      2018 Research-status Report
  • [Presentation] X-ray crystallographic snapshot of SufS-SufU complex in iron-sulfur cluster biosynthetic SUF-like machinery2018

    • Author(s)
      Takashi Fujishiro, Takuya Terahata, Kunichika Kouhei, Chihiro Maruyama, Kei Asai, Yasuhiro Takahashi
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 枯草菌の鉄硫黄クラスター生合成系の硫黄供給酵素複合体のX線結晶構造解析2018

    • Author(s)
      藤城 貴史・寺畑 拓也・國近 航平・横山 奈央・丸山 ちひろ・朝井 計・高橋 康弘
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 超好熱性細菌の鉄硫黄クラスター生合成系:IscS2‐IscU複合体のX線結晶構造解析2018

    • Author(s)
      國近 航平・藤城 貴史・山川 誠・平林 佳・岩永 朋子・福山 恵一・和田 啓・高橋 康弘
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Research-status Report
  • [Presentation] 構造情報を利用した枯草菌由来金属キレターゼ様タンパク質の性状解析2017

    • Author(s)
      藤城 貴史・朝井 計・高橋 康弘
    • Organizer
      2017年度生命科学系学会合同年次大会(ConBio2017)
    • Related Report
      2017 Research-status Report
  • [Presentation] The NIF machinery for iron-sulfur cluster biogenesis2017

    • Author(s)
      Takashi Fujishiro,Yasuhiro Takahashi
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Elucidation of catalytic mechanism of cysteine desulfurase NifS of NIF machinery in iron-sulfur cluster biogenesis2017

    • Author(s)
      中村亮裕、藤城貴史,高橋康弘
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Research-status Report
  • [Book] Bioorganometallic Chemistry2020

    • Author(s)
      Prof. Dr. Wolfgang Weigand and Dr. Ulf-Peter Apfel, Ed.
    • Total Pages
      435
    • Publisher
      De Gruyter
    • ISBN
      9783110496505
    • Related Report
      2019 Annual Research Report
  • [Remarks] システインから無機硫黄を取り出す酵素反応の瞬間を捉える!

    • URL

      http://www.saitama-u.ac.jp/topics_archives/2017-1215-0953-9.html

    • Related Report
      2017 Research-status Report

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Published: 2017-04-28   Modified: 2021-02-19  

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