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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Expression, purification, crystallization and preliminary crystallographic analysis of Leishmania mexicana phosphoglycerate mutase.
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Expression, purification, crystallization and preliminary crystallographic analysis of Leishmania mexicana phosphoglycerate mutase.

机译：表达,纯化,结晶初步晶体的分析利什曼虫墨西哥磷酸甘油酸酯变位酶。

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摘要

Bacterially expressed 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (iPGAM) from Leishmania mexicana with a six-His tag fused at its C-terminus was expressed from plasmid pET28a after IPTG induction in Escherichia coli cells and gave a yield of 20 mg of highly purified iPGAM per litre of cell culture. Crystals of the protein complexed with 3-phosphoglycerate were obtained by the hanging-drop method of vapour diffusion with PEG 4000 as the precipitating agent in the presence of cobalt chloride and diffracted synchrotron radiation to beyond 1.90 A. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 62.46, b = 72.27, c = 129.68 A. A model of Bacillus stearothermophilus iPGAM (33% identity) was used to provide an initial molecular-replacement solution. X-ray data to 2.05 A for the structure of L. mexicana iPGAM complexed with 2-phosphoglycerate have also been collected.
机译：细菌表达2, 3-bisphosphoglycerate-independent磷酸甘油酸酯变位酶(iPGAM)利什曼虫墨西哥six-His标签融合的糖基从质粒pET28a表示在IPTG诱导大肠杆菌细胞并给出一个产量20毫克的高纯度细胞培养的iPGAM每升。蛋白质包裹着3 -磷酸甘油酸通过蒸汽的悬滴法扩散与4000年沉淀的挂钩二氯化钴和代理衍射同步加速器辐射超过1.90答:晶体属于斜方晶系的空间集团P2(1) 2(1) 2(1),晶胞参数= 62.46, b = 72.27, c = 129.68。芽孢杆菌stearothermophilus iPGAM身份(33%)被用来提供一个初始的吗molecular-replacement解决方案。2.05结构的l .墨西哥iPGAM包裹着2-phosphoglycerate也收集。

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