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2019 年度 実績報告書

Adaptive optics approach for high-throughput, high-resolution cryo-electron microscopy of biological macromolecules and complexes

研究課題

研究課題/領域番号 19K21176
配分区分基金
研究機関東京大学

研究代表者

Danev Radostin  東京大学, 大学院医学系研究科(医学部), 教授 (50415931)

研究期間 (年度) 2019-04-01 – 2020-03-31
キーワードcryo-electron microscopy / molecular structure
研究実績の概要

The goal of this research was to improve the efficiency of electron cryo-microscopy (cryo-EM) experiments. Cryo-EM has become a fundamental method for high-resolution structure determination of biological macromolecules. However, to produce a high-quality result it requires acquisition of thousands of images from the sample. With previous methods this took several days. Cryo-EMs are expensive instruments. Reducing the time for data acquisition can speed up research and allow efficient use of research budgets. Previously, the typical throughput was ~1000 images/day. We developed and implemented a vastly improved “adaptive optics” method for automated image acquisition which increased the experimental throughput fivefold. The method uses optical shifting of the acquisition area, instead of the traditional mechanical stage movement, which saves significant amount of time. After development and testing of the new approach, we installed it on the main electron microscope of the new electron microscopy facility at the University of Tokyo. Our method became the standard automated data acquisition technique at the facility. We shared information about best practices and trained all experienced microscope users on its proper use. The data acquisition throughput increased to ~5500 images/day which enabled the acquisition of a complete dataset in one day. Consequently, the use efficiency of the facility was greatly improved, and this helped us and many other researchers to achieve outstanding high-resolution results in their molecular structural projects.

  • 研究成果

    (10件)

すべて 2020 2019

すべて 雑誌論文 (5件) (うち国際共著 5件、 査読あり 5件) 学会発表 (5件) (うち国際学会 2件、 招待講演 4件)

  • [雑誌論文] Activation of the GLP-1 receptor by a non-peptidic agonist2020

    • 著者名/発表者名
      Zhao Peishen、Liang Yi-Lynn、Belousoff Matthew J.、[14 co-authors]、Danev Radostin、Sexton Patrick M.、Wootten Denise
    • 雑誌名

      Nature

      巻: 577 ページ: 432~436

    • DOI

      10.1038/s41586-019-1902-z

    • 査読あり / 国際共著
  • [雑誌論文] Toward a Structural Understanding of Class B GPCR Peptide Binding and Activation2020

    • 著者名/発表者名
      Liang Yi-Lynn、Belousoff Matthew J.、Zhao Peishen、Koole Cassandra、Fletcher Madeleine M.、Truong Tin T.、Julita Villy、Christopoulos George、Xu H. Eric、Zhang Yan、Khoshouei Maryam、Christopoulos Arthur、Danev Radostin、Sexton Patrick M.、Wootten Denise
    • 雑誌名

      Molecular Cell

      巻: 77 ページ: 656~668.e5

    • DOI

      10.1016/j.molcel.2020.01.012

    • 査読あり / 国際共著
  • [雑誌論文] Structure and Dynamics of Adrenomedullin Receptors AM1 and AM2 Reveal Key Mechanisms in the Control of Receptor Phenotype by Receptor Activity-Modifying Proteins2020

    • 著者名/発表者名
      Liang Yi-Lynn、Belousoff Matthew J.、Fletcher Madeleine M.、Zhang Xin、Khoshouei Maryam、Deganutti Giuseppe、Koole Cassandra、Furness Sebastian G. B.、Miller Laurence J.、Hay Debbie L.、Christopoulos Arthur、Reynolds Christopher A.、Danev Radostin、Wootten Denise、Sexton Patrick M.
    • 雑誌名

      ACS Pharmacology & Translational Science

      巻: 3 ページ: 263~284

    • DOI

      10.1021/acsptsci.9b00080

    • 査読あり / 国際共著
  • [雑誌論文] Cryo-Electron Microscopy Methodology: Current Aspects and Future Directions2019

    • 著者名/発表者名
      Danev Radostin、Yanagisawa Haruaki、Kikkawa Masahide
    • 雑誌名

      Trends in Biochemical Sciences

      巻: 44 ページ: 837~848

    • DOI

      10.1016/j.tibs.2019.04.008

    • 査読あり / 国際共著
  • [雑誌論文] Improved applicability and robustness of fast cryo-electron tomography data acquisition2019

    • 著者名/発表者名
      Eisenstein Fabian、Danev Radostin、Pilhofer Martin
    • 雑誌名

      Journal of Structural Biology

      巻: 208 ページ: 107~114

    • DOI

      10.1016/j.jsb.2019.08.006

    • 査読あり / 国際共著
  • [学会発表] Phase plates in cryo-EM2019

    • 著者名/発表者名
      Radostin Danev
    • 学会等名
      EMBL Cryo-EM in Industry and Academia workshop
    • 国際学会 / 招待講演
  • [学会発表] Practical Performance of the Volta Phase Plate2019

    • 著者名/発表者名
      Radostin Danev
    • 学会等名
      Gordon Research Conference on Three Dimensional Electron Microscopy
    • 国際学会
  • [学会発表] Cryo-EM of Membrane Proteins With and Without Phase Plates2019

    • 著者名/発表者名
      Radostin Danev
    • 学会等名
      71st joint JSCB and PSSJ meeting
    • 招待講演
  • [学会発表] Cryo-EM of Membrane Receptors2019

    • 著者名/発表者名
      Radostin Danev
    • 学会等名
      92nd annual meeting of the Japanese biochemical society
    • 招待講演
  • [学会発表] Performance Contributions of Energy Filtering and Phase Plates for Single Particle Analysis of Membrane Proteins2019

    • 著者名/発表者名
      Radostin Danev
    • 学会等名
      NIPS EM Workshop 2019
    • 招待講演

URL: 

公開日: 2021-01-27  

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