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Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser
Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables essentially radiation-damage-free macromolecular structure determination using microcrystals that are too small for synchrotron studies. However, SFX experiments often require large amounts of sample in order to c...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8562661/ https://www.ncbi.nlm.nih.gov/pubmed/34804544 http://dx.doi.org/10.1107/S2052252521008046 |
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author | Nass, Karol Bacellar, Camila Cirelli, Claudio Dworkowski, Florian Gevorkov, Yaroslav James, Daniel Johnson, Philip J. M. Kekilli, Demet Knopp, Gregor Martiel, Isabelle Ozerov, Dmitry Tolstikova, Alexandra Vera, Laura Weinert, Tobias Yefanov, Oleksandr Standfuss, Jörg Reiche, Sven Milne, Christopher J. |
author_facet | Nass, Karol Bacellar, Camila Cirelli, Claudio Dworkowski, Florian Gevorkov, Yaroslav James, Daniel Johnson, Philip J. M. Kekilli, Demet Knopp, Gregor Martiel, Isabelle Ozerov, Dmitry Tolstikova, Alexandra Vera, Laura Weinert, Tobias Yefanov, Oleksandr Standfuss, Jörg Reiche, Sven Milne, Christopher J. |
author_sort | Nass, Karol |
collection | PubMed |
description | Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables essentially radiation-damage-free macromolecular structure determination using microcrystals that are too small for synchrotron studies. However, SFX experiments often require large amounts of sample in order to collect highly redundant data where some of the many stochastic errors can be averaged out to determine accurate structure-factor amplitudes. In this work, the capability of the Swiss X-ray free-electron laser (SwissFEL) was used to generate large-bandwidth X-ray pulses [Δλ/λ = 2.2% full width at half-maximum (FWHM)], which were applied in SFX with the aim of improving the partiality of Bragg spots and thus decreasing sample consumption while maintaining the data quality. Sensitive data-quality indicators such as anomalous signal from native thaumatin micro-crystals and de novo phasing results were used to quantify the benefits of using pink X-ray pulses to obtain accurate structure-factor amplitudes. Compared with data measured using the same setup but using X-ray pulses with typical quasi-monochromatic XFEL bandwidth (Δλ/λ = 0.17% FWHM), up to fourfold reduction in the number of indexed diffraction patterns required to obtain similar data quality was achieved. This novel approach, pink-beam SFX, facilitates the yet underutilized de novo structure determination of challenging proteins at XFELs, thereby opening the door to more scientific breakthroughs. |
format | Online Article Text |
id | pubmed-8562661 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-85626612021-11-18 Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser Nass, Karol Bacellar, Camila Cirelli, Claudio Dworkowski, Florian Gevorkov, Yaroslav James, Daniel Johnson, Philip J. M. Kekilli, Demet Knopp, Gregor Martiel, Isabelle Ozerov, Dmitry Tolstikova, Alexandra Vera, Laura Weinert, Tobias Yefanov, Oleksandr Standfuss, Jörg Reiche, Sven Milne, Christopher J. IUCrJ Research Papers Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables essentially radiation-damage-free macromolecular structure determination using microcrystals that are too small for synchrotron studies. However, SFX experiments often require large amounts of sample in order to collect highly redundant data where some of the many stochastic errors can be averaged out to determine accurate structure-factor amplitudes. In this work, the capability of the Swiss X-ray free-electron laser (SwissFEL) was used to generate large-bandwidth X-ray pulses [Δλ/λ = 2.2% full width at half-maximum (FWHM)], which were applied in SFX with the aim of improving the partiality of Bragg spots and thus decreasing sample consumption while maintaining the data quality. Sensitive data-quality indicators such as anomalous signal from native thaumatin micro-crystals and de novo phasing results were used to quantify the benefits of using pink X-ray pulses to obtain accurate structure-factor amplitudes. Compared with data measured using the same setup but using X-ray pulses with typical quasi-monochromatic XFEL bandwidth (Δλ/λ = 0.17% FWHM), up to fourfold reduction in the number of indexed diffraction patterns required to obtain similar data quality was achieved. This novel approach, pink-beam SFX, facilitates the yet underutilized de novo structure determination of challenging proteins at XFELs, thereby opening the door to more scientific breakthroughs. International Union of Crystallography 2021-09-23 /pmc/articles/PMC8562661/ /pubmed/34804544 http://dx.doi.org/10.1107/S2052252521008046 Text en © Karol Nass et al. 2021 https://creativecommons.org/licenses/by/4.0/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. |
spellingShingle | Research Papers Nass, Karol Bacellar, Camila Cirelli, Claudio Dworkowski, Florian Gevorkov, Yaroslav James, Daniel Johnson, Philip J. M. Kekilli, Demet Knopp, Gregor Martiel, Isabelle Ozerov, Dmitry Tolstikova, Alexandra Vera, Laura Weinert, Tobias Yefanov, Oleksandr Standfuss, Jörg Reiche, Sven Milne, Christopher J. Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title | Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title_full | Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title_fullStr | Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title_full_unstemmed | Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title_short | Pink-beam serial femtosecond crystallography for accurate structure-factor determination at an X-ray free-electron laser |
title_sort | pink-beam serial femtosecond crystallography for accurate structure-factor determination at an x-ray free-electron laser |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8562661/ https://www.ncbi.nlm.nih.gov/pubmed/34804544 http://dx.doi.org/10.1107/S2052252521008046 |
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