Novel approach to phasing proteins: derivatization by short cryo-soaking with halides
Novel approach to phasing proteins: derivatization by short cryo-soaking with halides
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DOI：
10.1107/s0907444999016352
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发表时间：
2000-02-01
影响因子：
2.2
通讯作者：
Rajashankar, KR
中科院分区：
文献类型：
--
作者：
Dauter, Z;Dauter, M;Rajashankar, KR
快速（小于1分钟）浸泡的蛋白质晶体在含有溴或碘阴离子的低温溶液导致这些异常散射纳入到有序的溶剂区域周围的蛋白质分子。这些卤素阴离子提供了一种方便的方法，通过其异常散射信号的相位：溴化物使用多波长异常色散（MAD）和溴化物和/或碘化物使用单波长异常色散（SAD）或单一同晶置换与异常散射（SIRAS）方法。这种方法已经在四种不同的蛋白质上成功地进行了测试，并已用于解决分子量为30 kDa的新蛋白质的结构。
A quick (less than 1 min) soak of protein crystals in a cryo-solution containing bromide or iodide anions leads to incorporation of these anomalous scatterers into the ordered solvent region around the protein molecules. These halide anions provide a convenient way of phasing through their anomalous scattering signal: bromides using multiwavelength anomalous dispersion (MAD) and bromides and/or iodides using single-wavelength anomalous dispersion (SAD) or single isomorphous replacement with anomalous scattering (SIRAS) methods. This approach has been tested successfully on four different proteins and has been used to solve the structure of a new protein of molecular weight 30 kDa.