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Establishment of technique for supersaturation control in liquid cell transmission electron microscopy and its application to protein crystallization

Research Project

Project/Area Number 20H02580
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 28040:Nanobioscience-related
Research InstitutionHokkaido University

Principal Investigator

Yamazaki Tomoya  北海道大学, 低温科学研究所, 特任助教 (50735032)

Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2022: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2021: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2020: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Keywords透過型電子顕微鏡 / 溶液セル / タンパク質 / 過飽和度 / 誘電泳動 / 試料冷却TEMホルダー / 放射線分解 / 結晶化 / 氷 / タンパク質結晶 / その場観察
Outline of Research at the Start

溶液からタンパク質が結晶化するとき、結晶の種となる結晶核が生成する。この過程を核生成と呼ぶ。近年、その前後の過程において、従来観測されてこなかった現象(結晶核の生成に必要な前駆物質の生成など)が見つかりつつある。本研究ではまず、これらの現象を捉えることが可能な溶液セル透過型電子顕微法において、結晶化の駆動力を制御するための技術を確立する。そして核生成前後の描像解明にアプローチする。

Outline of Final Research Achievements

To elucidate the crystallization process of protein crystals, it is necessary to observe the crystallization process with high temporal and spatial resolution. Such observation can be performed with a liquid-cell transmission electron microscopy. In order to perform efficient crystallization in the liquid cell, techniques to control supersaturation in the liquid cell were investigated. Three techniques were developed: radiolysis combined with anti-solvent, dielectrophoresis; specimen cooling. The application of these techniques to protein crystallization was investigated.

Academic Significance and Societal Importance of the Research Achievements

結晶化の物理・化学的メカニズムの解明は長年に渡り取り組まれている学術的に重要な課題である。また結晶化は、創薬に関わるタンパク質分子の構造解析、産業に関わる半導体などの材料開発、地球の熱収支に関わる雲の生成などの基礎であり、社会的に重要な現象である。これを調べるのに有効な溶液セル透過型電子顕微鏡法において、結晶化を能動的に生じさせる手法を開発した。これにより結晶化の瞬間を捉えることが容易になり、結晶化の理解が飛躍的に進展することが期待できる。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (18 results)

All 2023 2022 2021 2020

All Journal Article (7 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 2 results) Presentation (11 results) (of which Int'l Joint Research: 3 results)

  • [Journal Article] Feasibility of control of particle assembly by dielectrophoresis in liquid-cell transmission electron microscopy2022

    • Author(s)
      Yamazaki Tomoya、Niinomi Hiromasa、Kimura Yuki
    • Journal Title

      Microscopy

      Volume: 71 Issue: 4 Pages: 231-237

    • DOI

      10.1093/jmicro/dfac021

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Is dielectrophoresis effective for increasing local concentration of particles in liquid-cell transmission electron microscopy?2022

    • Author(s)
      Yamazaki Tomoya、Niinomi Hiromasa、Katsuno Hiroyasu、Hosseinkhannazer Hooman、Daigle Eric、Kimura Yuki
    • Journal Title

      Microscopy and Microanalysis

      Volume: 28 Issue: S1 Pages: 1934-1936

    • DOI

      10.1017/s1431927622007565

    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Journal Article] Radiolysis-Induced Crystallization of Sodium Chloride in Acetone by Electron Beam Irradiation2021

    • Author(s)
      Yamazaki Tomoya、Kimura Yuki
    • Journal Title

      Microscopy and Microanalysis

      Volume: - Issue: 3 Pages: 1-7

    • DOI

      10.1017/s1431927621000179

    • NAID

      120007171676

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid Ortho-to-para Nuclear Spin Conversion of H2 on a Silicate Dust Surface2021

    • Author(s)
      Tsuge M., Namiyoshi T., Furuya K., Yamazaki T., Kouchi A., Watanabe N.
    • Journal Title

      The Astrophysical Journal

      Volume: 908 Issue: 2 Pages: 234-234

    • DOI

      10.3847/1538-4357/abd9c0

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In-situ water-immersion experiments on amorphous silicates in the MgO?SiO2 system: implications for the onset of aqueous alteration in primitive meteorites2021

    • Author(s)
      Igami Yohei、Tsuchiyama Akira、Yamazaki Tomoya、Matsumoto Megumi、Kimura Yuki
    • Journal Title

      Geochimica et Cosmochimica Acta

      Volume: 293 Pages: 86-102

    • DOI

      10.1016/j.gca.2020.10.023

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] High-Density Liquid Water at a Water?Ice Interface2020

    • Author(s)
      Niinomi Hiromasa、Yamazaki Tomoya、Nada Hiroki、Hama Tetsuya、Kouchi Akira、Okada Junpei T.、Nozawa Jun、Uda Satoshi、Kimura Yuki
    • Journal Title

      The Journal of Physical Chemistry Letters

      Volume: 11 Issue: 16 Pages: 6779-6784

    • DOI

      10.1021/acs.jpclett.0c01907

    • NAID

      120007140366

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Time-Resolved Observation of Evolution of Amyloid-β Oligomer with Temporary Salt Crystals2020

    • Author(s)
      Nakajima Kichitaro、Yamazaki Tomoya、Kimura Yuki、So Masatomo、Goto Yuji、Ogi Hirotsugu
    • Journal Title

      The Journal of Physical Chemistry Letters

      Volume: 11 Issue: 15 Pages: 6176-6184

    • DOI

      10.1021/acs.jpclett.0c01487

    • NAID

      120007148352

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Presentation] ペルチェ式冷却TEM ホルダーと溶液セルを用いた氷結晶その場観察2023

    • Author(s)
      山﨑智也,屋嶋悠河,勝野弘康, 宮崎裕也,權堂貴志,木村勇気
    • Organizer
      JpGU 2023
    • Related Report
      2022 Annual Research Report
  • [Presentation] 溶液セルとペルチェ式冷却TEMホルダーを用いた氷結晶のその場観察2023

    • Author(s)
      山﨑智也、屋嶋悠河、勝野弘康、宮崎裕也、權堂貴志、木村勇気
    • Organizer
      The 79th Annual Meeting of the Japanese Society of Microscopy
    • Related Report
      2022 Annual Research Report
  • [Presentation] Liquid‐Cell Transmission Electron Microscopy of ice crystals and bubble behaviors in the ice2023

    • Author(s)
      Tomoya Yamazaki, Yuga Yashima, Hiroyasu Katsuno, Hiroya Miyazaki, Takashi Gondo, Yuki Kimura
    • Organizer
      PCI-2023
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 誘電泳動による粒子挙動の溶液セルTEMその場観察2022

    • Author(s)
      山﨑智也, 新家寛正, 勝野弘康, 木村勇気
    • Organizer
      The 78th Annual Meeting of the Japanese Society of Microscopy
    • Related Report
      2022 Annual Research Report
  • [Presentation] 溶液セル透過型電子顕微鏡法におけるサブミクロンサイズの粒子の誘電泳動2022

    • Author(s)
      山﨑智也, 新家寛正, 勝野弘康, 木村勇気
    • Organizer
      JpGU 2022
    • Related Report
      2022 Annual Research Report
  • [Presentation] Is dielectrophoresis effective for increasing local concentration of particles in liquid-cell transmission electron microscopy?2022

    • Author(s)
      Tomoya Yamazaki, Hiromasa Niinomi, Hiroyasu Katsuno, Hooman Hosseinkhannazer, Eric Daigle, Yuki Kimura
    • Organizer
      M&M 2022, on-demand
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ペルチェ式冷却TEMホルダーと溶液セルを用いた結晶化その場観察2022

    • Author(s)
      山﨑智也,屋嶋悠河,宮崎裕也,權堂貴志,木村勇気
    • Organizer
      JCCG-51
    • Related Report
      2022 Annual Research Report
  • [Presentation] Control of particles by AC field and its applicability in LC-TEM for observing crystallization and dissolution processes2021

    • Author(s)
      Tomoya Yamazaki, Hiromasa Niinomi, Yuki Kimura
    • Organizer
      JpGU 2021
    • Related Report
      2021 Annual Research Report
  • [Presentation] 溶液セルTEMにおける結晶化条件制御方法の検討2021

    • Author(s)
      山﨑智也, 木村勇気
    • Organizer
      日本顕微鏡学会 第77回学術講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] Supersaturation control for observation of crystallization in liquid-cell transmission electron microscopy2021

    • Author(s)
      Tomoya Yamazaki, Yuki Kimura
    • Organizer
      Pacifichem 2021
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Radiolysis-induced crystallization process of salt in liquid cell transmission electron microscopy2020

    • Author(s)
      Yamazaki Tomoya、Kimura Yuki
    • Organizer
      JpGU-AGU Joint Meeting 2020
    • Related Report
      2020 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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