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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Crystallization, dehydration and experimental phasing of WbdD, a bifunctional kinase and methyltransferase from Escherichia coli O9a
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Crystallization, dehydration and experimental phasing of WbdD, a bifunctional kinase and methyltransferase from Escherichia coli O9a

机译：结晶、脱水和实验世界献血日的定相双官能团的激酶从大肠杆菌O9a甲基转移酶

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摘要

WbdD is a bifunctional kinase/methyltransferase that is responsible for regulation of lipopolysaccharide O antigen polysaccharide chain length in Escherichia coli serotype O9a. Solving the crystal structure of this protein proved to be a challenge because the available crystals belonging to space group I23 only diffracted to low resolution (>95% of the crystals diffracted to resolution lower than 4 ? and most only to 8 ?) and were non-isomorphous, with changes in unit-cell dimensions of greater than 10%. Data from a serendipitously found single native crystal that diffracted to 3.0 ? resolution were non-isomorphous with a lower (3.5 ?) resolution selenomethionine data set. Here, a strategy for improving poor (3.5 ? resolution) initial phases by density modification and cross-crystal averaging with an additional 4.2 ? resolution data set to build a crude model of WbdD is desribed. Using this crude model as a mask to cut out the 3.5 ? resolution electron density yielded a successful molecular-replacement solution of the 3.0 ? resolution data set. The resulting map was used to build a complete model of WbdD. The hydration status of individual crystals appears to underpin the variable diffraction quality of WbdD crystals. After the initial structure had been solved, methods to control the hydration status of WbdD were developed and it was thus possible to routinely obtain high-resolution diffraction (to better than 2.5 ? resolution). This novel and facile crystal-dehydration protocol may be useful for similar challenging situations.
机译：世界献血日是一个双官能激酶/甲基转移酶负责监管脂多糖O抗原多糖链长度在大肠杆菌血清型O9a。事实证明这种蛋白质的晶体结构是一个挑战,因为可用的晶体只属于空间群I23衍射低分辨率(> 95%的晶体衍射分辨率低于4 ??)和non-isomorphous,变化单胞尺寸大于10%。从一个偶然发现单独的本地晶体衍射到3.0吗?non-isomorphous较低(3.5 ?)决议硒代蛋氨酸数据集。改善贫困(3.5吗?通过修改和cross-crystal密度平均一个额外的4.2吗?数据集建立的粗糙模型世界献血日描述。我把3.5吗?一个成功的molecular-replacement解决3.0吗?世界献血日的被用来建立一个完整的模型。水合状态的个体晶体出现支撑变量衍射质量世界献血日晶体。被解决,方法控制水化世界献血日的状态是这样可能经常获得高分辨率衍射(比2.5更好?这本小说和肤浅crystal-dehydration协议可以用于类似的挑战的情况。

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