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Title: Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography.
Authors: Hasegawa, Kazuya
Yamashita, Keitaro
Murai, Tomohiro
Nuemket, Nipawan
Hirata, Kunio
Ueno, Go
Ago, Hideo
Nakatsu, Toru
Kumasaka, Takashi
Yamamoto, Masaki
Author's alias: 長谷川, 和也
山下, 恵太郎
村井, 智洋
ニパワン, ヌアムケット
平田, 邦生
上野, 剛
吾郷, 日出夫
中津, 亨
熊坂, 崇
山本, 雅貴
Keywords: serial synchrotron crystallography
mercury single-wavelength anomalous diffraction
radiation damage
Issue Date: Jan-2017
Publisher: International Union of Crystallography
Journal title: Journal of synchrotron radiation
Volume: 24
Issue: Pt. 1
Start page: 29
End page: 41
Abstract: Serial crystallography, in which single-shot diffraction images are collected, has great potential for protein microcrystallography. Although serial femtosecond crystallography (SFX) has been successfully demonstrated, limited beam time prevents its routine use. Inspired by SFX, serial synchrotron crystallography (SSX) has been investigated at synchrotron macromolecular crystallography beamlines. Unlike SFX, the longer exposure time of milliseconds to seconds commonly used in SSX causes radiation damage. However, in SSX, crystals can be rotated during the exposure, which can achieve efficient coverage of the reciprocal space. In this study, mercury single-wavelength anomalous diffraction (Hg-SAD) phasing of the luciferin regenerating enzyme (LRE) was performed using serial synchrotron rotation crystallography. The advantages of rotation and influence of dose on the data collected were evaluated. The results showed that sample rotation was effective for accurate data collection, and the optimum helical rotation step depended on multiple factors such as multiplicity and partiality of reflections, exposure time per rotation angle and the contribution from background scattering. For the LRE microcrystals, 0.25° was the best rotation step for the achievable resolution limit, whereas a rotation step larger than or equal to 1° was favorable for Hg-SAD phasing. Although an accumulated dose beyond 1.1 MGy caused specific damage at the Hg site, increases in resolution and anomalous signal were observed up to 3.4 MGy because of a higher signal-to-noise ratio.
Description: 微小タンパク質結晶からの効率的な構造解析法 : 凍結した試料を回転させる「SS-ROX法」を確立、汎用化へ. 京都大学プレスリリース. 2017-01-12.
Rights: This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
URI: http://hdl.handle.net/2433/217787
DOI(Published Version): 10.1107/S1600577516016362
PubMed ID: 28009544
Related Link: http://www.kyoto-u.ac.jp/ja/research/research_results/2016/161202_3.html
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