使用融合蛋白获得适合X射线分析的晶体：包含DNA复制相关元件结合因子DREF的DNA结合结构域的GST融合蛋白的结晶。
Use of a fusion protein to obtain crystals suitable for X-ray analysis: crystallization of a GST-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF.
作者信息
Kuge M, Fujii Y, Shimizu T, Hirose F, Matsukage A, Hakoshima T
机构信息
Department of Molecular Biology, Nara Institute of Science and Technology (NAIST), Japan.
出版信息
Protein Sci. 1997 Aug;6(8):1783-6. doi: 10.1002/pro.5560060822.
Abstract
Crystals of glutathione-S-transferase (GST)-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF, were obtained under crystallization conditions similar to those for GST. Preliminary X-ray crystallographic analysis revealed that crystals of the GST-fused protein belong to space group P6(1)22 or P6(5)22 with unit cell dimensions a = b = 140.4 A, c = 93.5 A and gamma = 120 degrees, having one molecule in the crystallographic asymmetric unit. The crystals diffract to 2.5 A resolution. The cell dimensions are related to those of GST crystals thus far reported. Crystallization of the DNA-binding domain that was cleaved from the fused protein by thrombin was also carried out using several methods under numerous conditions, but efforts to produce well-ordered large crystals were unsuccessful. A possible application of GST-fusion proteins for small target proteins or domains to obtain crystals suitable for X-ray structure determination is proposed.
摘要
在与谷胱甘肽 - S - 转移酶（GST）相似的结晶条件下，获得了含有DNA复制相关元件结合因子DREF的DNA结合结构域的GST融合蛋白晶体。初步的X射线晶体学分析表明，GST融合蛋白晶体属于空间群P6(1)22或P6(5)22，晶胞参数为a = b = 140.4 Å，c = 93.5 Å，γ = 120°，在晶体学不对称单元中有一个分子。这些晶体的衍射分辨率为2.5 Å。其晶胞参数与迄今报道的GST晶体的晶胞参数相关。还使用多种方法在众多条件下对通过凝血酶从融合蛋白上切割下来的DNA结合结构域进行了结晶，但未能成功获得排列良好的大晶体。提出了GST融合蛋白在小的靶蛋白或结构域上的一种可能应用，以获得适合X射线结构测定的晶体。
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Use of a fusion protein to obtain crystals suitable for X-ray analysis: crystallization of a GST-fused protein containing the DNA-binding domain of DNA replication-related element-binding factor, DREF.使用融合蛋白获得适合X射线分析的晶体：包含DNA复制相关元件结合因子DREF的DNA结合结构域的GST融合蛋白的结晶。
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