Tracking Protein Motions using Serial Femtosecond Crystallography with X-Ray Free-Electron Laser

Mizohata, Eiichi; Nango, Eriko; Tosha, Takehiko et al.

Current Protocols, 2025, 5(9), e70212

アクセス数:72件（2026-02-16 10:17 集計）

固定URL: https://hdl.handle.net/11094/102967

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タイトル
Tracking Protein Motions using Serial Femtosecond Crystallography with X-Ray Free-Electron Laser
著者
NRID
1000090571183 1000090571183
researchmap permalink
read0068597 read0068597
Scopus Author ID
9742902400 9742902400
著者名
Mizohata, Eiichi
NRID
1000090376947 1000090376947
著者名
Nango, Eriko
NRID
1000000548993 1000000548993
著者名
Tosha, Takehiko
NRID
1000060452330 1000060452330
著者名
Iwata, So
NRID
1000090392878 1000090392878
著者名
Kubo, Minoru
キーワード等
molecular movie
protein motion
SACLA
structure–function relationship
time-resolved analysis
内容
Mizohata E., Nango E., Tosha T., et al. Tracking Protein Motions using Serial Femtosecond Crystallography with X-Ray Free-Electron Laser. Current Protocols 5, e70212 (2025); https://doi.org/10.1002/cpz1.70212.
抄録
Since the birth of biochemistry, researchers have investigated the structure–function relationship of a wide variety of proteins. However, until recently, when X-ray free-electron lasers (XFELs) became available, it was not possible to visualize the motion of proteins from moment to moment with excellent temporal and spatial resolution. Here, we introduce practical methods to visualize protein motions at room temperature using serial femtosecond crystallography (SFX) using XFELs. With the development of this technology, it will be possible to visualize the entire reaction mechanism of many proteins in the future. We first outline a streamlined microcrystallization workflow for hen egg-white lysozyme, enabling rapid detector calibration and data-collection optimization. Next, we present a rotational seeding approach refined on copper-containing nitrite reductase that yields homogeneous microcrystals suitable for high-resolution SFX and readily adaptable to other challenging targets. Finally, we describe a time-resolved strategy combining microcrystals of fungal nitric-oxide reductase with photolabile caged substrates and synchronized UV triggering, capturing catalytic intermediates on the millisecond timescale. Together, these procedures enable investigators to progress from preparing samples to capturing dynamic structural snapshots. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Microcrystallization of lysozyme. Basic Protocol 2: Microcrystallization of copper-containing nitrite reductase. Basic Protocol 3: Time-resolved serial femtosecond crystallography.
出版者
John Wiley and Sons Inc
収録物名
Current Protocols
巻(号)
5(9)
ページ
e70212
刊行年月
2025-09-23
言語
英語
ハンドルURL
https://hdl.handle.net/11094/102967
eISSN
26911299
関連情報 (isIdenticalTo)
DOI (出版社版)
https://doi.org/10.1002/cpz1.70212
アクセス権
open access
権利情報
This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ
Version of Record
カテゴリ
学術雑誌論文 Journal Article