[注] 参考文献133件を含む16頁
構造生物学は、健康と疾患における分子機械の機能を解明する手掛かりをもたらすが、その究極の課題は、高解像度の構造の詳細と、機能に不可欠な動的な構造変化の双方を捉えることにある。
X線結晶構造解析（以下、X線解析）とクライオ電子顕微鏡（以下、クライオ電顕）といった高解像度手法は、主として、静的コンフォメーションの集合体を研究するために使用されてきたが、動的遷移状態を捉えることにも使用できる。
ハンブルグを拠点とする研究チームが今回、動的コンフォメーションの捉え方に焦点を当て、時間分解構造生物学における最新の戦略と進歩を概説する。X線解析とクライオ電顕における時間分解サンプル調製とデリバリーに関する最近の技術開発について解説し、これらの技術が分子レベルおよび原子レベルの生物学への理解を深めるためにどのように相互に利益をもたらすことができるかを探る。
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- MAIN
- Fig. 1: Biology is dynamic, spanning processes from subfemtosecond electronic transitions to second-long conformational changes or reactions.
- Vitrification methods for cryo-EM samples for trapping MD
- Fig. 2: Different sample preparation techniques for time-resolved cryo-EM reaction initiation.
- Table 1 Comparison of cryo-EM vitrification methods, sorted by vitrification time reported, and selection of published time-resolved X-ray crystallography methods, sorted by shortest time delay reported
- Traditional preparation of cryo-EM samples
- Freezing a dynamic system using traditional sample preparation methods
- The microfluidic ‘mix-and-spray’ method
- ‘On-grid’ mixing approaches
- Time-resolved cryo-EM sample preparation on microsecond to millisecond timescales
- The melt-and-freeze method
- The pump-and-freeze method
- Cryo-EM data analysis of heterogeneous datasetsTime-resolved sample preparation in X-ray science
- Time-resolved sample preparation in X-ray science
- Sample preparation for time-resolved serial crystallography
- Fig. 3: Sample delivery for XFEL and SSX experiments follows six concepts.
- Fig. 4: Light-triggered structural dynamics of DNA repair captured by time-resolved crystallography.
- Analysis of XFEL single-particle imaging data and parallels with cryo-EM
- Fig. 5: Parallels between cryo-EM (top row) and XFEL SPI (bottom row) single-particle workflows.
- Future time-resolved opportunities and cross-fertilization of ideas
- References
[出典] Review "Advancing time-resolved structural biology: latest strategies in cryo-EM and X-ray crystallography" Banari A [..] Seuring C. Nat Methods. 2025-05-01. https://doi.org/10.1038/s41592-025-02659-6 [著者所属] Centre for Structural Systems Biology (Germany), The Hamburg Centre for Ultrafast Imaging, Center for Free-Electron Laser Science, U Hamburg, Department of Structural Cell Biology (Leibniz Institute of Virology), U Medical Center Hamburg-Eppendorf, Deutsches Elektronen-Synchrotron DESY, Laboratory of Molecular Biophysics (Uppsala U, Sweden), Division of Structural Biology (U Oxford, UK)
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