Dauter Miroslawa, Dauter Zbigniew
Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne National Laboratory, Argonne, IL, USA.
Methods Mol Biol. 2007;364:149-58. doi: 10.1385/1-59745-266-1:149.
A short soak of protein crystals in cryosolution containing bromides or iodides leads to incorporation of these ions into the ordered solvent shell around the protein surface. The halide ions display significant anomalous signal, bromides in the vicinity of the absorption edge at 0.92 A, and iodides at longer wavelengths, e.g., provided by the copper sources. Bromides can, therefore, be used through multiwavelength anomalous diffraction or single-wavelength anomalous diffraction (SAD) techniques and iodides through SAD or multiple isomorphous replacement (MIRAS) phasing. The halide cryosoaking approach involves very little preparative effort and offers a rapid and simple way of solving novel protein crystal structures.
将蛋白质晶体在含有溴化物或碘化物的冷冻溶液中短暂浸泡,会导致这些离子掺入蛋白质表面周围有序的溶剂层中。卤离子显示出显著的反常信号,溴化物在0.92 Å吸收边缘附近,碘化物在更长波长处,例如由铜源提供。因此,溴化物可通过多波长反常衍射或单波长反常衍射(SAD)技术使用,碘化物可通过SAD或多同晶置换(MIRAS)相位法使用。卤化物冷冻浸泡方法几乎不需要制备工作,并提供了一种快速简单的解析新蛋白质晶体结构的方法。