Long-wavelength native-SAD phasing : opportunities and challenges

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2019
Autor:innen
Basu, Shibom
Olieric, Vincent
Leonarski, Filip
Matsugaki, Naohiro
Kawano, Yoshiaki
Takashi, Tomizaki
Huang, Chia-Ying
Yamada, Yusuke
Wang, Meitian
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IUCrJ. 2019, 6, pp. 373-386. eISSN 2052-2525. Available under: doi: 10.1107/S2052252519002756
Zusammenfassung

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 (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 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.

Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
570 Biowissenschaften, Biologie
Schlagwörter
single-wavelength anomalous dispersion; native-SAD phasing; Se/S-SAD; UV-laser cutting; crystal shaping; spherical crystals;absorption correction; anomalous scattering factor; structure determination
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ISO 690BASU, Shibom, Vincent OLIERIC, Filip LEONARSKI, Naohiro MATSUGAKI, Yoshiaki KAWANO, Tomizaki TAKASHI, Chia-Ying HUANG, Yusuke YAMADA, Kay DIEDERICHS, Meitian WANG, 2019. Long-wavelength native-SAD phasing : opportunities and challenges. In: IUCrJ. 2019, 6, pp. 373-386. eISSN 2052-2525. Available under: doi: 10.1107/S2052252519002756
BibTex
@article{Basu2019Longw-45811,
  year={2019},
  doi={10.1107/S2052252519002756},
  title={Long-wavelength native-SAD phasing : opportunities and challenges},
  volume={6},
  journal={IUCrJ},
  pages={373--386},
  author={Basu, Shibom and Olieric, Vincent and Leonarski, Filip and Matsugaki, Naohiro and Kawano, Yoshiaki and Takashi, Tomizaki and Huang, Chia-Ying and Yamada, Yusuke and Diederichs, Kay and Wang, Meitian}
}
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