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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Generating isomorphous heavy-atom derivatives by a quick-soak method. Part II: phasing of new structures.
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Generating isomorphous heavy-atom derivatives by a quick-soak method. Part II: phasing of new structures.

机译：产生同形的重原子衍生物quick-soak方法。结构。

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A quick-soak method has been applied to generate de novo heavy-atom phasing to solve two new protein structures, a type II transforming growth factor beta receptor (TBRII) and a natural killer cell receptor-ligand complex, NKG2D-ULBP3. In the case of TBRII, a crystal derivatized for only 10 min in saturated HgCl(2) provided adequate phasing for structure determination. Comparison between HgCl(2) derivatives generated by 10 min soaking and by 12 h soaking revealed similar phasing statistics. The shorter soak, however, resulted in a derivative more isomorphous to the native than the longer soak as judged by changes in the unit-cell parameter a upon derivatization as well as by the quality of a combined SIRAS electron-density map. In the case of the NKG2D-ULBP3 structure, all overnight soaks in heavy-atom solutions resulted in crystal lattice disorder and only the quick soaks preserved diffraction. Despite fragile lattice packing, the quick-soaked K(2)PtCl(4) derivative was isomorphous with the native crystal and the electron-density map calculated from combined SIR and MAD phases is better than that calculated from MAD phases alone. Combined with mass-spectrometry-assisted solution heavy-atom derivative screening and the use of synchrotron radiation, the quick-soak derivatization has the potential to transform the time-consuming conventional heavy-atom search into a real-time 'on-the-fly' derivatization process that will benefit high-throughput structural genomics.
机译：quick-soak方法已经应用于生成新创重原子逐步解决两个新的蛋白质结构,II型转变增长因子β受体(TBRII)和自然杀伤细胞receptor-ligand复杂,NKG2D-ULBP3。TBRII,水晶derivatized仅为10最小饱和HgCl(2)提供足够了定相结构的决心。HgCl(2)之间所产生的衍生品10分钟浸泡和12 h显示相似定相统计数据。导致衍生更多的同形本机比浸泡时间越长根据变化对衍生化的晶胞参数以及联合SIRAS的质量电子密度图。NKG2D-ULBP3结构,所有隔夜浸泡重原子导致晶格的解决方案障碍,只有快速浸泡保存衍射。quick-soaked K(2)竞购(4)导数同晶型与本机水晶和电子密度图计算出结合先生和疯狂的阶段比计算单从疯狂的阶段。mass-spectrometry-assisted解决重原子导数筛查和同步加速器的使用辐射,quick-soak衍生的潜力将耗时传统的重原子搜索实时“动态”的衍生化过程高通量的结构基因组学中获益。

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