Benjamin Stieglitz, Katrin Rittinger, Lesley F Haire
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引用次数: 0
摘要
人 SHARPIN 的 N 端片段在大肠杆菌中重组表达、纯化并结晶。通过使用二维网格筛一步优化种子稀释和蛋白质浓度,获得了适合 X 射线衍射的晶体。晶体属于原始四方空间群 P4(3)2(1)2,单位晶胞参数 a = b = 61.55,c = 222.81 Å。从 100 K 的原生和硒代蛋氨酸蛋白质晶体中收集了完整的数据集,分辨率分别为 2.6 和 2.0 Å。
Crystallization of SHARPIN using an automated two-dimensional grid screen for optimization.
An N-terminal fragment of human SHARPIN was recombinantly expressed in Escherichia coli, purified and crystallized. Crystals suitable for X-ray diffraction were obtained by a one-step optimization of seed dilution and protein concentration using a two-dimensional grid screen. The crystals belonged to the primitive tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 61.55, c = 222.81 Å. Complete data sets were collected from native and selenomethionine-substituted protein crystals at 100 K to 2.6 and 2.0 Å resolution, respectively.
期刊介绍:
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