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Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography

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 c...

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Autores principales: Hasegawa, Kazuya, Yamashita, Keitaro, Murai, Tomohiro, Nuemket, Nipawan, Hirata, Kunio, Ueno, Go, Ago, Hideo, Nakatsu, Toru, Kumasaka, Takashi, Yamamoto, Masaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5182019/
https://www.ncbi.nlm.nih.gov/pubmed/28009544
http://dx.doi.org/10.1107/S1600577516016362
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author Hasegawa, Kazuya
Yamashita, Keitaro
Murai, Tomohiro
Nuemket, Nipawan
Hirata, Kunio
Ueno, Go
Ago, Hideo
Nakatsu, Toru
Kumasaka, Takashi
Yamamoto, Masaki
author_facet Hasegawa, Kazuya
Yamashita, Keitaro
Murai, Tomohiro
Nuemket, Nipawan
Hirata, Kunio
Ueno, Go
Ago, Hideo
Nakatsu, Toru
Kumasaka, Takashi
Yamamoto, Masaki
author_sort Hasegawa, Kazuya
collection PubMed
description 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.
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spelling pubmed-51820192017-01-10 Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography Hasegawa, Kazuya Yamashita, Keitaro Murai, Tomohiro Nuemket, Nipawan Hirata, Kunio Ueno, Go Ago, Hideo Nakatsu, Toru Kumasaka, Takashi Yamamoto, Masaki J Synchrotron Radiat Radiation Damage 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. International Union of Crystallography 2017-01-01 /pmc/articles/PMC5182019/ /pubmed/28009544 http://dx.doi.org/10.1107/S1600577516016362 Text en © Kazuya Hasegawa et al. 2017 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Radiation Damage
Hasegawa, Kazuya
Yamashita, Keitaro
Murai, Tomohiro
Nuemket, Nipawan
Hirata, Kunio
Ueno, Go
Ago, Hideo
Nakatsu, Toru
Kumasaka, Takashi
Yamamoto, Masaki
Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title_full Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title_fullStr Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title_full_unstemmed Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title_short Development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
title_sort development of a dose-limiting data collection strategy for serial synchrotron rotation crystallography
topic Radiation Damage
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5182019/
https://www.ncbi.nlm.nih.gov/pubmed/28009544
http://dx.doi.org/10.1107/S1600577516016362
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