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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...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2019
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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. |
format | Online Article Text |
id | pubmed-6503925 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
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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