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  Long-wavelength native-SAD phasing: opportunities and challenges

Basu, S., Olieric, V., Leonarski, F., Matsugaki, N., Kawano, Y., Takashi, T., et al. (2019). Long-wavelength native-SAD phasing: opportunities and challenges. IUCrJ, 6(3), 373-386. doi:10.1107/S2052252519002756.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0003-E7FC-D Version Permalink: http://hdl.handle.net/21.11116/0000-0003-E7FD-C
Genre: Journal Article

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 Creators:
Basu, Shibom1, Author
Olieric, Vincent1, Author
Leonarski, Filip1, Author
Matsugaki, Naohiro1, Author
Kawano, Yoshiaki1, Author
Takashi, Tomizaki1, Author
Huang, Chia-Ying1, Author
Yamada, Yusuke1, Author
Vera, Laura1, Author
Olieric, Natacha1, Author
Basquin, Jerome2, Author              
Wojdyla, Justyna A.1, Author
Bunk, Oliver1, Author
Diederichs, Kay1, Author
Yamamoto, Masaki1, Author
Wang, Meitian1, Author
Affiliations:
1external, ou_persistent22              
2Conti, Elena / Structural Cell Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565144              

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Free keywords: SOFT X-RAYS; DATA-COLLECTION; MACROMOLECULAR CRYSTALLOGRAPHY; ANOMALOUS DIFFRACTION; ATOMIC-RESOLUTION; PROTEIN CRYSTALS; STRUCTURAL BASIS; ROUTINE USE; OXIDASE; FAMILYChemistry; Crystallography; Materials Science; single-wavelength anomalous dispersion; native-SAD phasing; Se/S-SAD; UV-laser cutting; crystal shaping; spherical crystals; absorption correction; anomalous scattering factor; structure determination;
 Abstract: 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 angstrom is demonstrated for native-SAD phasing on a 266 kDa multiprotein-ligand tubulin complex (T2R-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 angstrom is reported for native SAD. The potential of laser-shaping technology and other challenges for an optimized native-SAD experiment at wavelengths >3 angstrom are discussed.

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Language(s): eng - English
 Dates: 2019
 Publication Status: Published in print
 Pages: 14
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: ISI: 000466861200006
DOI: 10.1107/S2052252519002756
 Degree: -

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Title: IUCrJ
Source Genre: Journal
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Publ. Info: Chester CH1 2HU, England : International Union of Crystallography (IUCr)
Pages: - Volume / Issue: 6 (3) Sequence Number: - Start / End Page: 373 - 386 Identifier: ISSN: 2052-2525
CoNE: https://pure.mpg.de/cone/journals/resource/2052-2525