Molecular cloning, expression, purification and crystallographic analysis of zebrafish THEM2.
Molecular cloning, expression, purification and crystallographic analysis of zebrafish THEM2.
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DOI：
10.1107/s1744309112043813
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发表时间：
2012-12
期刊：
影响因子：
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通讯作者：
Han Li;F. Gao;Shanshan Yu;M. Jia;W. Gong
中科院分区：
文献类型：
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作者：
Han Li;F. Gao;Shanshan Yu;M. Jia;W. Gong
硫酯酶超家族成员 2 (THEM2) 对于哺乳动物细胞的细胞增殖至关重要。它属于热狗折叠硫酯酶超家族，在体外催化酰基辅酶A硫酯键的水解。在本研究中，来自斑马鱼的 THEM2 蛋白 (fTHEM2) 在大肠杆菌中表达，并通过 Ni 亲和层析和凝胶过滤层析进行纯化。 fTHEM2晶体是通过坐滴蒸气扩散法以PEG 10 000作为沉淀剂获得的。使用同步加速器辐射源收集分辨率为 1.80 Å 的 X 射线衍射数据。该晶体属于单斜晶系C2空间群，晶胞参数a=77.1，b=74.4，c=96.6 Å，β=93.7°。
Thioesterase superfamily member 2 (THEM2) is essential for cell proliferation of mammalian cells. It belongs to the hotdog-fold thioesterase superfamily and catalyzes the hydrolysis of the thioester bonds of acyl-CoA in vitro. In this study, THEM2 protein from zebrafish (fTHEM2) was expressed in Escherichia coli and purified by Ni-affinity and gel-filtration chromatography. fTHEM2 crystals were obtained using the sitting-drop vapour-diffusion method with PEG 10 000 as precipitant. X-ray diffraction data were collected to 1.80 Å resolution using a synchrotron-radiation source. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a=77.1, b=74.4, c=96.6 Å, β=93.7°.