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Development of novel biosensor probes capable of sensing dynamic changes in intracellular sodium ion concentration

Research Project

Project/Area Number 22K19274
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionOsaka University

Principal Investigator

Minamino Tohru  大阪大学, 大学院生命機能研究科, 准教授 (20402993)

Project Period (FY) 2022-06-30 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2023: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords細菌 / 蛋白質 / 遺伝学 / 感染症
Outline of Research at the Start

同一のゲノム情報を持つにもかかわらず、機能的に分化した細胞群が様々な役割分担をしてバイオフィルムを維持している。バイオフィルムに生息する大部分の細胞では運動機能は抑制されているが、ごく稀にべん毛運動する細胞群が存在する。バイオフィルムに生息する細胞と同様にATPaseリング複合体が働かないサルモネラ変異体を用いた研究から、ナトリウムイオンが不均一な細胞集団を出現させる重要なシグナルである可能性が示唆された。この仮説を検証するため、バイオフィルム内部に生息する細菌の細胞内ナトリウムの動的変化を計測できる新規のバイオセンサープローブの開発を目指す。

Outline of Final Research Achievements

The MotPS complex acts as a sodium ion-driven stator unit in the Bacillus subtilis flagellar motor and binds to the peptidoglycan layer via the peptidoglycan-binding domain of MotS (MotS-PGB) to become an active stator unit. When the external sodium ion concentration exceeds about 130 mM, sodium ions bind directly to MotS-PGB and induce the disorder-to-order transition of MotS-PGB. When sodium ions are depleted, MotS-PGB becomes unstructured, causing rapid dissociation of the MotPS complex from the motor. Such conformational changes of MotS-PGB autonomously regulates the number of active stator units around the flagellar motor. In this study, we show that the Asp-70 and Glu-76 residues of MotS are sodium ion binding sites and that the region from Gln-68 to Glu-117 of MotS undergoes a reversible conformational change in the presence of sodium ions.

Academic Significance and Societal Importance of the Research Achievements

従来の蛍光色素を用いた細胞内ナトリウムイオン濃度の動的時間変化の測定は、定量的な解析が困難であるとともに、蛍光色素による細胞毒性が大きな問題となっている。本研究で同定したナトリウムセンサーはナトリウムイオンの濃度変化に迅速に応答できる非侵襲性バイオセンサープローブの開発に利用できる。このようなバイオセンサープローブは様々な生物種の細胞に発現させることができるため、ナトリウムイオンが関与する様々な生命現象の解明に貢献することが期待される。

Report

(3 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • Research Products

    (21 results)

All 2024 2023 2022 Other

All Journal Article (8 results) (of which Peer Reviewed: 8 results,  Open Access: 5 results) Presentation (11 results) (of which Invited: 2 results) Book (1 results) Remarks (1 results)

  • [Journal Article] FliH and FliI help FlhA bring strict order to flagellar protein export in Salmonella2024

    • Author(s)
      Kinoshita Miki、Minamino Tohru、Uchihashi Takayuki、Namba Keiichi
    • Journal Title

      Communications Biology

      Volume: 7 Issue: 1 Pages: 336-336

    • DOI

      10.1038/s42003-024-06081-0

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structure, Assembly, and Function of Flagella Responsible for Bacterial Locomotion2023

    • Author(s)
      Minamino Tohru、Kinoshita Miki
    • Journal Title

      EcoSal Plus

      Volume: 11 Issue: 1

    • DOI

      10.1128/ecosalplus.esp-0011-2023

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Frontiers of microbial movement research2023

    • Author(s)
      Minamino Tohru、Nakane Daisuke、Nakamura Shuichi、Kiyama Hana、Morimoto Yusuke V.、Miyata Makoto
    • Journal Title

      Biophysics and Physicobiology

      Volume: 20 Issue: 3 Pages: n/a

    • DOI

      10.2142/biophysico.bppb-v20.0033

    • ISSN
      2189-4779
    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Loss of Flagella-Related Genes Enables a Nonflagellated, Fungal-Predating Bacterium To Strengthen the Synthesis of an Antifungal Weapon2023

    • Author(s)
      Xiong Dan、Yang Zixiang、He Xueting、He Weimei、Shen Danyu、Wang Lu、Lin Long、Murero Aprodisia、Minamino Tohru、Shao Xiaolong、Qian Guoliang
    • Journal Title

      Microbiology Spectrum

      Volume: 11 Issue: 1

    • DOI

      10.1128/spectrum.04149-22

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Purification of the Transmembrane Polypeptide Channel Complex of the Salmonella Flagellar Type III Secretion System2023

    • Author(s)
      Kinoshita Miki、Namba Keiichi、Minamino Tohru
    • Journal Title

      Methods in Molecular Biology

      Volume: 2646 Pages: 3-15

    • DOI

      10.1007/978-1-0716-3060-0_1

    • ISBN
      9781071630594, 9781071630600
    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Measurements of the Ion Channel Activity of the Transmembrane Stator Complex in the Bacterial Flagellar Motor2023

    • Author(s)
      Morimoto Yusuke V.、Minamino Tohru
    • Journal Title

      Methods in molecular biology

      Volume: 2646 Pages: 83-94

    • DOI

      10.1007/978-1-0716-3060-0_8

    • ISBN
      9781071630594, 9781071630600
    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] High-Resolution Rotation Assay of the Bacterial Flagellar Motor Near Zero Loads Using a Mutant Having a Rod-Like Straight Hook2023

    • Author(s)
      Nakamura Shuichi、Minamino Tohru
    • Journal Title

      Methods in Molecular Biology

      Volume: 2646 Pages: 125-131

    • DOI

      10.1007/978-1-0716-3060-0_11

    • ISBN
      9781071630594, 9781071630600
    • Related Report
      2022 Research-status Report
    • Peer Reviewed
  • [Journal Article] Activation mechanism of the bacterial flagellar dual-fuel protein export engine2022

    • Author(s)
      Minamino Tohru、Kinoshita Miki、Morimoto Yusuke V.、Namba Keiichi
    • Journal Title

      Biophysics and Physicobiology

      Volume: 19 Issue: 0 Pages: n/a

    • DOI

      10.2142/biophysico.bppb-v19.0046

    • ISSN
      2189-4779
    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] サルモネラ直線型べん毛繊維L型とR型の高分解能構造比較2024

    • Author(s)
      牧野文信, Kasim Waraichi, 木下実紀, 宮田知子, 南野徹, 難波啓一
    • Organizer
      べん毛研究交流会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 海洋ビブリオ菌の極べん毛モーターFliFG融合タンパク質からなるMSリングの構造と性状2024

    • Author(s)
      竹川宜宏, 錦野達郎, 岸川淳一, 廣瀬未果, 木下実紀, 小嶋誠司, 南野徹, 内橋貴之, 今田勝巳, 本間道夫
    • Organizer
      べん毛研究交流会
    • Related Report
      2023 Annual Research Report
  • [Presentation] べん毛輸送ATPase FliI 6量体の構造変化2024

    • Author(s)
      竹川宜宏, 碓井亜瑳子, 多治見祐希, 内橋貴之, 木下実紀, 南野徹, 今田勝巳
    • Organizer
      べん毛研究交流会
    • Related Report
      2023 Annual Research Report
  • [Presentation] バクテリアべん毛で働くプロトン駆動力の計測 (Measurement of proton motive force acting on bacterial flagella)2023

    • Author(s)
      森本雄祐, 南野徹
    • Organizer
      日本バイオイメージング学会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] Proton-protein antiporter mechanism in the type III secretion system of the bacterial flagellum (細菌べん毛の III 型分泌システムにおけるプロトン-タンパク質アンチポーター機構)2023

    • Author(s)
      Minamino T, Kinoshita M, Namba K.
    • Organizer
      日本生物物理学会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] CryoEM structure of the polypeptide channel complex of the bacterial flagellar type III secretion system (べん毛 III 型分泌装置のポリペプチドチャネル複合体のクライオ電子顕微鏡構造)2023

    • Author(s)
      Kinoshita M, Miyata T, Makino F, Kato T, Imada K, Namba K, Minamino T.
    • Organizer
      日本生物物理学会
    • Related Report
      2023 Annual Research Report
  • [Presentation] CryoEM structure of the ATPase ring complex of the flagellar type III export apparatus(べん毛III型輸送ATPase複合体のcryoEM構造)2023

    • Author(s)
      Usui A, Kinoshita M, Tajimi Y, Uchihashi T, Minamino T, Takekawa N, Imada K.
    • Organizer
      日本生物物理学会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Development of a novel membrane voltage sensor based on the bacterial flagellar type III secretion system (バクテリアべん毛Ⅲ型輸送装置を利用した新規膜電位センサーの開発)2023

    • Author(s)
      Sakata K, Minamino T, Morimoto YV.
    • Organizer
      日本生物物理学会
    • Related Report
      2023 Annual Research Report
  • [Presentation] べん毛T3SS ATPaseの構造変化2023

    • Author(s)
      碓井亜瑳子, 多治見祐希, 内橋貴之, 木下実紀, 南野徹, 竹川宜宏, 今田勝巳
    • Organizer
      生命分子科学研究会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Role of the cytoplasmic ATPase complex in export switching of the flagellar protein export apparatus.2023

    • Author(s)
      Minamino T, Kinoshita M, Namba K.
    • Organizer
      日本細菌学会
    • Related Report
      2022 Research-status Report
  • [Presentation] サルモネラIII型輸送装置のポリペプチドチャネル複合体の形成2023

    • Author(s)
      木下実紀, 宮田知子, 牧野文信, 難波啓一, 南野徹.
    • Organizer
      べん毛研究交流会
    • Related Report
      2022 Research-status Report
  • [Book] Bacterial and Archaeal Motility - Methods and Protocols.2023

    • Author(s)
      Minamino T , Miyata M, Namba K.
    • Total Pages
      405
    • Publisher
      Humana Press.
    • Related Report
      2022 Research-status Report
  • [Remarks]

    • URL

      https://rd.iai.osaka-u.ac.jp/ja/2ce577c0931a0530.html

    • Related Report
      2022 Research-status Report

URL: 

Published: 2022-07-05   Modified: 2025-01-30  

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