Pushing the limits of sulfur SAD phasing: de novo structure solution of the N-terminal domain of the ectodomain of HCV E1.
Pushing the limits of sulfur SAD phasing: de novo structure solution of the N-terminal domain of the ectodomain of HCV E1.
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DOI：
10.1107/s139900471401339x
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发表时间：
2014-08
期刊：
影响因子：
--
通讯作者：
Stuart DI
中科院分区：
文献类型：
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作者：
El Omari K;Iourin O;Kadlec J;Fearn R;Hall DR;Harlos K;Grimes JM;Stuart DI
硫SAD定相方法成功地用于确定HCV E1的N-末端结构域的低分辨率衍射晶体的结构，通过结合来自32个晶体的数据。单波长S原子反常色散（S-SAD）是一种不需要重原子掺入或硒代甲硫氨酸衍生化的精密定相方法。然而，这种技术受到通常X射线波长信号缺乏的限制，需要非常精确地测量异常差异。在这里，数据收集和结构的解决方案的N-末端结构域的胞外域的HCV E1从晶体衍射非常弱的报告。通过结合来自32个晶体的数据，可以求解硫子结构并以7 μ m分辨率计算初始图，并且在密度修改和相扩展之后，使用更高分辨率的天然数据集可以实现3.5 μ m分辨率的模型构建。
The sulfur SAD phasing method was successfully used to determine the structure of the N-terminal domain of HCV E1 from low-resolution diffracting crystals by combining data from 32 crystals. Single-wavelength anomalous dispersion of S atoms (S-SAD) is an elegant phasing method to determine crystal structures that does not require heavy-atom incorporation or selenomethionine derivatization. Nevertheless, this technique has been limited by the paucity of the signal at the usual X-ray wavelengths, requiring very accurate measurement of the anomalous differences. Here, the data collection and structure solution of the N-terminal domain of the ectodomain of HCV E1 from crystals that diffracted very weakly is reported. By combining the data from 32 crystals, it was possible to solve the sulfur substructure and calculate initial maps at 7 Å resolution, and after density modication and phase extension using a higher resolution native data set to 3.5 Å resolution model building was achievable.