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Long-wavelength native-SAD phasing: opportunities and challenges

Native single-wavelength anomalous dispersion (SAD) is an attractive experimental phasing technique as it exploits weak anomalous signals from intrinsic light scatterers (Z < 20). The anomalous signal of sulfur in particular, is enhanced at long wavelengths, however the absorption of diffracted X...

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Autores principales: Basu, Shibom, Olieric, Vincent, Leonarski, Filip, Matsugaki, Naohiro, Kawano, Yoshiaki, Takashi, Tomizaki, Huang, Chia-Ying, Yamada, Yusuke, Vera, Laura, Olieric, Natacha, Basquin, Jerome, Wojdyla, Justyna A., Bunk, Oliver, Diederichs, Kay, Yamamoto, Masaki, Wang, Meitian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6503925/
https://www.ncbi.nlm.nih.gov/pubmed/31098019
http://dx.doi.org/10.1107/S2052252519002756
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author Basu, Shibom
Olieric, Vincent
Leonarski, Filip
Matsugaki, Naohiro
Kawano, Yoshiaki
Takashi, Tomizaki
Huang, Chia-Ying
Yamada, Yusuke
Vera, Laura
Olieric, Natacha
Basquin, Jerome
Wojdyla, Justyna A.
Bunk, Oliver
Diederichs, Kay
Yamamoto, Masaki
Wang, Meitian
author_facet Basu, Shibom
Olieric, Vincent
Leonarski, Filip
Matsugaki, Naohiro
Kawano, Yoshiaki
Takashi, Tomizaki
Huang, Chia-Ying
Yamada, Yusuke
Vera, Laura
Olieric, Natacha
Basquin, Jerome
Wojdyla, Justyna A.
Bunk, Oliver
Diederichs, Kay
Yamamoto, Masaki
Wang, Meitian
author_sort Basu, Shibom
collection PubMed
description Native single-wavelength anomalous dispersion (SAD) is an attractive experimental phasing technique as it exploits weak anomalous signals from intrinsic light scatterers (Z < 20). The anomalous signal of sulfur in particular, is enhanced at long wavelengths, however the absorption of diffracted X-rays owing to the crystal, the sample support and air affects the recorded intensities. Thereby, the optimal measurable anomalous signals primarily depend on the counterplay of the absorption and the anomalous scattering factor at a given X-ray wavelength. Here, the benefit of using a wavelength of 2.7 over 1.9 Å is demonstrated for native-SAD phasing on a 266 kDa multiprotein-ligand tubulin complex (T(2)R-TTL) and is applied in the structure determination of an 86 kDa helicase Sen1 protein at beamline BL-1A of the KEK Photon Factory, Japan. Furthermore, X-ray absorption at long wavelengths was controlled by shaping a lysozyme crystal into spheres of defined thicknesses using a deep-UV laser, and a systematic comparison between wavelengths of 2.7 and 3.3 Å is reported for native SAD. The potential of laser-shaping technology and other challenges for an optimized native-SAD experiment at wavelengths >3 Å are discussed.
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spelling pubmed-65039252019-05-16 Long-wavelength native-SAD phasing: opportunities and challenges Basu, Shibom Olieric, Vincent Leonarski, Filip Matsugaki, Naohiro Kawano, Yoshiaki Takashi, Tomizaki Huang, Chia-Ying Yamada, Yusuke Vera, Laura Olieric, Natacha Basquin, Jerome Wojdyla, Justyna A. Bunk, Oliver Diederichs, Kay Yamamoto, Masaki Wang, Meitian IUCrJ Research Papers Native single-wavelength anomalous dispersion (SAD) is an attractive experimental phasing technique as it exploits weak anomalous signals from intrinsic light scatterers (Z < 20). The anomalous signal of sulfur in particular, is enhanced at long wavelengths, however the absorption of diffracted X-rays owing to the crystal, the sample support and air affects the recorded intensities. Thereby, the optimal measurable anomalous signals primarily depend on the counterplay of the absorption and the anomalous scattering factor at a given X-ray wavelength. Here, the benefit of using a wavelength of 2.7 over 1.9 Å is demonstrated for native-SAD phasing on a 266 kDa multiprotein-ligand tubulin complex (T(2)R-TTL) and is applied in the structure determination of an 86 kDa helicase Sen1 protein at beamline BL-1A of the KEK Photon Factory, Japan. Furthermore, X-ray absorption at long wavelengths was controlled by shaping a lysozyme crystal into spheres of defined thicknesses using a deep-UV laser, and a systematic comparison between wavelengths of 2.7 and 3.3 Å is reported for native SAD. The potential of laser-shaping technology and other challenges for an optimized native-SAD experiment at wavelengths >3 Å are discussed. International Union of Crystallography 2019-04-01 /pmc/articles/PMC6503925/ /pubmed/31098019 http://dx.doi.org/10.1107/S2052252519002756 Text en © Shibom Basu et al. 2019 http://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.http://creativecommons.org/licenses/by/4.0/
spellingShingle Research Papers
Basu, Shibom
Olieric, Vincent
Leonarski, Filip
Matsugaki, Naohiro
Kawano, Yoshiaki
Takashi, Tomizaki
Huang, Chia-Ying
Yamada, Yusuke
Vera, Laura
Olieric, Natacha
Basquin, Jerome
Wojdyla, Justyna A.
Bunk, Oliver
Diederichs, Kay
Yamamoto, Masaki
Wang, Meitian
Long-wavelength native-SAD phasing: opportunities and challenges
title Long-wavelength native-SAD phasing: opportunities and challenges
title_full Long-wavelength native-SAD phasing: opportunities and challenges
title_fullStr Long-wavelength native-SAD phasing: opportunities and challenges
title_full_unstemmed Long-wavelength native-SAD phasing: opportunities and challenges
title_short Long-wavelength native-SAD phasing: opportunities and challenges
title_sort long-wavelength native-sad phasing: opportunities and challenges
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6503925/
https://www.ncbi.nlm.nih.gov/pubmed/31098019
http://dx.doi.org/10.1107/S2052252519002756
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