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Analysis of ion dynamics in organelles by using targetable probes

Research Project

Project/Area Number 18H02102
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 37030:Chemical biology-related
Research InstitutionTohoku University

Principal Investigator

Mizukami Shin  東北大学, 多元物質科学研究所, 教授 (30420433)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000)
Fiscal Year 2020: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2019: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2018: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
Keywords蛍光プローブ / Mg2+ / Zn2+ / 蛋白質ラベル化 / 定量 / 蛋白質ラベル化技術 / Mg2+イオン / Zn2+イオン / 局在化 / オルガネラ / 蛍光イメージング / 金属イオン / マグネシウム / 亜鉛 / 定量的イメージング / オルガネラ内 / イメージング
Outline of Final Research Achievements

In this study, quantitative imaging techniques of Mg2+ and Zn2+ dynamics in organelles were developed. By integrating a highly selective Mg2+ probe, which is unaffected by changes in intracellular Ca2+ concentration, with protein labeling technology, we developed a technique for selectively visualizing Mg2+ in organelles and showed that the free Mg2+ concentration in the nucleus fluctuates during cell division. We also developed a localizable zinc quantification probe and found that the level of Zn2+ in the Golgi is about several tens of nM. Furthermore, we developed new probes with different dissociation constants and succeeded in quantitative imaging of free Zn2+ in the cytoplasm, nucleus, Golgi apparatus, endoplasmic reticulum and mitochondria.

Academic Significance and Societal Importance of the Research Achievements

細胞機能を理解するためには、分子の存在だけではなく、分子がどこにどれくらい存在するか(時空間的かつ定量的な情報)を知ることが重要である。化学的な高選択性プローブを細胞内局所へ局在させることによって、蛋白質プローブや低分子プローブでは得られない時空間分解能を持った金属イオン定量が可能になった。特に、亜鉛イオンの詳細な細胞内濃度マップに関する知見は、亜鉛イオンに関連する疾患機構の解明や治療薬の開発に繋がることが期待できる。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (42 results)

All 2021 2020 2019 2018 Other

All Int'l Joint Research (1 results) Journal Article (4 results) (of which Peer Reviewed: 4 results,  Open Access: 1 results) Presentation (31 results) (of which Int'l Joint Research: 12 results,  Invited: 4 results) Book (2 results) Remarks (3 results) Patent(Industrial Property Rights) (1 results)

  • [Int'l Joint Research] Thomas-Jefferson University(米国)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Protocol for synthesis and use of a turn-on fluorescent probe for quantifying labile Zn2+ in the Golgi apparatus in live cells2021

    • Author(s)
      Kowada Toshiyuki、Watanabe Tomomi、Liu Rong、Mizukami Shin
    • Journal Title

      STAR Protocols

      Volume: 2 Issue: 2 Pages: 100395-100395

    • DOI

      10.1016/j.xpro.2021.100395

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Optical Manipulation of Subcellular Protein Translocation Using a Photoactivatable Covalent Labeling System2021

    • Author(s)
      Kowada Toshiyuki、Arai Keisuke、Yoshimura Akimasa、Matsui Toshitaka、Kikuchi Kazuya、Mizukami Shin
    • Journal Title

      Angewandte Chemie International Edition

      Volume: 60 Issue: 20 Pages: 11378-11383

    • DOI

      10.1002/anie.202016684

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Quantitative Imaging of Labile Zn2+ in the Golgi Apparatus Using a Localizable Small-Molecule Fluorescent Probe2020

    • Author(s)
      Kowada Toshiyuki、Watanabe Tomomi、Amagai Yuta、Liu Rong、Yamada Momo、Takahashi Hiroto、Matsui Toshitaka、Inaba Kenji、Mizukami Shin
    • Journal Title

      Cell Chemical Biology

      Volume: 27 Issue: 12 Pages: 1521-1531

    • DOI

      10.1016/j.chembiol.2020.09.003

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Light-wavelength-based Quantitative Control of Dihydrofolate Reductase Activity Using Photochromic Isostere of Inhibitor2019

    • Author(s)
      Takato Mashita, Toshiyuki Kowada, Hiroto Takahashi, Toshitaka Matsui, Shin Mizukami
    • Journal Title

      ChemBioChem

      Volume: 印刷中 Issue: 11 Pages: 1382-1386

    • DOI

      10.1002/cbic.201800816

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] Quantitative Mapping of Cellular Labile Zn2+ via Development of Hybrid Fluorescent Probes2021

    • Author(s)
      劉熔, 小和田 俊行, 松井 敏高, 水上 進
    • Organizer
      日本化学会 第101春季年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 細胞機能を探索するための 光応答性プローブの開発2021

    • Author(s)
      水上進
    • Organizer
      日本薬学会第141年会一般シンポジウム「タンパク質高速分子動画に向けた光薬理学の新展開」
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 蛋白質ラベル化に基づく 細胞内分子の計測と制御2021

    • Author(s)
      水上進
    • Organizer
      新学術領域「生命金属科学」領域会議第1回地方巡業(仙台)
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 細胞小器官局在化蛍光プローブの開発と細胞内遊離亜鉛の定量イメージング2020

    • Author(s)
      小和田俊行, 渡邊朝美, 天貝佑太, 劉 熔, 山田 桃, 高橋泰人, 松井敏高, 稲葉謙次, 水上 進
    • Organizer
      第29回日本バイオイメージング学会学術集会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Quantitative Mapping of Organellar Labile Zn2+ via the Development of Hybrid Fluorescent Probes2020

    • Author(s)
      劉 熔, 小和田 俊行, 松井 敏高, 水上 進
    • Organizer
      第14回バイオ関連化学シンポジウム
    • Related Report
      2020 Annual Research Report
  • [Presentation] Quantitative imaging analysis of labile Zn2+ in intracellular organelles via the development of hybrid fluorescent probe2020

    • Author(s)
      劉熔, 小和田俊行, 渡邊朝美, 松井俊高, 水上進
    • Organizer
      理学・生命科学研究科合同シンポジウム 2020
    • Related Report
      2019 Annual Research Report
  • [Presentation] Quantitative Mapping of Cellular Labile Zn2+ via Development of Hybrid Fluorescent Probes2020

    • Author(s)
      劉熔, 小和田 俊行, 渡邊 朝美, 松井 敏高, 水上 進
    • Organizer
      日本化学会 第100春季年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 蛋白質ラベル化技術を用いた生体分子の可視化と機能制御2019

    • Author(s)
      水上 進
    • Organizer
      32回XFEL構造生物学ミーティング
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Quantitative analysis of free zinc ion concentration in living cells using novel targetable fluorescent probe2019

    • Author(s)
      Tomomi Watanabe, Toshiyuki Kowada, Rong Liu, Toshitaka Matsui, Shin Mizukami
    • Organizer
      15th International Symposium on Applied Bioinorganic Chemistry
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of Fluorescent Probes for Visualization and Quantification of Zn2+ in Organelles2019

    • Author(s)
      Rong Liu, Toshiyuki Kowada, Tomomi Watanabe, Toshitaka Matsui, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2019
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 細胞内小器官における亜鉛イオン濃度解析のための蛍光プローブの開発2019

    • Author(s)
      小和田 俊行, 劉 熔, 渡邊 朝美, 松井 敏高, 水上 進
    • Organizer
      第13回バイオ関連化学シンポジウム
    • Related Report
      2019 Annual Research Report
  • [Presentation] Development of chemical probes for investigating biomolecular dynamics in living cells2019

    • Author(s)
      Shin Mizukami
    • Organizer
      10th RSC-CSJ Joint Symposium
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Quantitative Mapping of Cellular Labile Zn2+ via Development of Hybrid Fluorescent Probes2019

    • Author(s)
      Rong Liu, Toshiyuki Kowada, Tomomi Watanabe, Toshitaka Matsui, Shin Mizukami
    • Organizer
      第19回東北大学多元物質科学研究所研究発表会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Development of fluorescent probes for visualization and quantitative analysis of free zinc ion in intracellular organelles2019

    • Author(s)
      Toshiyuki Kowada, Tomomi Watanabe, Rong Liu, Toshitaka Matsui, Shin Mizukami
    • Organizer
      Chemical Biology and Physiology Conference 2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] 蛋白質ラベル化技術を利用した蛋白質二量体化の光制御2019

    • Author(s)
      小和田 俊行、荒 井 啓介、吉村 彰真、松井 敏高、菊地 和也、水上 進
    • Organizer
      日本化学会第99春季年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] タンパク質への結合を可逆的に光制御可能なメトトレキセート誘導体の開発2019

    • Author(s)
      間下 貴斗, 小和田 俊行, 高橋 泰人, 松井 敏高, 水上 進
    • Organizer
      日本化学会第99春季年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 生体内オルガネラにおける亜鉛イオンの可視化2019

    • Author(s)
      渡邊朝美, 小和田俊行, 松井敏高, 水上 進
    • Organizer
      日本化学会第99春季年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Development of fluorescent probe for visualization and quantitative analysis of zinc ion in intracellular organelles2019

    • Author(s)
      Toshiyuki Kowada, Tomomi Watanabe, Toshitaka Matsui, Shin Mizukami
    • Organizer
      The 2nd Symposium for World Leading Research Centers
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of light-switchable protein-protein interaction system for protein degradation2019

    • Author(s)
      Toshiyuki Kowada, Toshitaka Matsui, Shin Mizukami
    • Organizer
      The 1st International Symposium on Chemical Communication (ISCC2019)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 可逆的光応答性薬剤の開発2018

    • Author(s)
      間下 貴斗, 小和田 俊行, 高橋 泰人, 松井 敏高, 水上 進
    • Organizer
      第29回 万有仙台シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] 細胞内オルガネラにおける亜鉛イオン動態の可視化2018

    • Author(s)
      渡邊 朝美, 小和田 俊行, 松井 敏高, 水上 進
    • Organizer
      第28回金属の関与する生体関連反応シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] 細胞内局所のpH定量を目的としたイメージングプローブの開発2018

    • Author(s)
      伊藤 理紗, 小和田 俊行, 松井 敏高, 水上 進
    • Organizer
      第12回バイオ関連化学シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] 光でタンパク質への結合を可逆的に制御可能なリガンドの開発2018

    • Author(s)
      間下 貴斗, 小和田 俊行, 高橋 泰人, 松井 敏高, 水上 進
    • Organizer
      第12回バイオ関連化学シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] 細胞内オルガネラにおける亜鉛イオンの可視化2018

    • Author(s)
      渡邊朝美, 小和田俊行, 松井敏高, 水上進
    • Organizer
      第18回多元研研究発表会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Live-cell fluorescence imaging of zinc ions using localizable small-molecule probe2018

    • Author(s)
      Toshiyuki Kowada, Tomomi Watanabe, Toshitaka Matsui, Shin Mizukami
    • Organizer
      5th CWRU-Tohoku Joint Workshop
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of fluorescent probes for real-time and quantitative measurement of local pH changes in cell2018

    • Author(s)
      Risa Ito, Toshiyuki Kowada, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of Photofunctional Tools Based on Chemistry and Biology2018

    • Author(s)
      Shin Mizukami
    • Organizer
      The 2nd International Symposium on Chemical Communication
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Development of fluorescent probes for visualization of zinc ion in intracellular organelles2018

    • Author(s)
      Tomomi Watanabe, Toshiyuki Kowada, Toshitaka Matsui, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Photo-switchable Dihydrofolate Reductase (DHFR) Inhibitor2018

    • Author(s)
      Takato Mashita, Toshiyuki Kowada, Hiroto Takahashi, Toshitaka Matsui, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of fluorescent probe for visualizing intracellular Mg2+ dynamics2018

    • Author(s)
      Keisuke Wakabayashi, Toshiyuki Kowada, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of self-assembling peptide for controlling aggregation state by light2018

    • Author(s)
      Yuri Hashimoto, Toshiyuki Kowada, Shin Mizukami
    • Organizer
      Tohoku University's Chemistry Summer School 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Book] イメージングの選び方・使い方100+(監修:原田慶恵、永井健治)2018

    • Author(s)
      小和田俊行・水上進
    • Publisher
      羊土社
    • ISBN
      4758103755
    • Related Report
      2018 Annual Research Report
  • [Book] 有機フッ素化合物の最新動向 (監修:今野勉)2018

    • Author(s)
      水上進
    • Publisher
      シーエムシー出版
    • ISBN
      9784781313375
    • Related Report
      2018 Annual Research Report
  • [Remarks] 東北大学多元物質科学研究所細胞機能分子化学研究分野

    • URL

      http://www2.tagen.tohoku.ac.jp/lab/mizukami/

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
  • [Remarks] プレスリリース:細胞小器官内の遊離亜鉛イオンの定量技術を開発

    • URL

      http://www2.tagen.tohoku.ac.jp/lab/news_press/20200930/

    • Related Report
      2020 Annual Research Report
  • [Remarks]

    • URL

      http://www2.tagen.tohoku.ac.jp/lab/mizukami/%e7%a0%94%e7%a9%b6%e5%86%85%e5%ae%b9/

    • Related Report
      2018 Annual Research Report
  • [Patent(Industrial Property Rights)] 光応答性化合物2018

    • Inventor(s)
      水上進,間下貴斗,小和田俊行,松井敏高
    • Industrial Property Rights Holder
      国立大学法人東北大学
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2018-165991
    • Filing Date
      2018
    • Related Report
      2018 Annual Research Report

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Published: 2018-04-23   Modified: 2022-01-27  

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