Overexpression, purification, crystallization and preliminary X-ray crystallographic studies of a proline-specific aminopeptidase from Aneurinibacillus sp. strain AM-1.
Overexpression, purification, crystallization and preliminary X-ray crystallographic studies of a proline-specific aminopeptidase from Aneurinibacillus sp. strain AM-1.
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DOI：
10.1107/s1744309106047543
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发表时间：
2006-12
期刊：
影响因子：
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通讯作者：
M. Akioka;H. Nakano;Aya Horikiri;Y. Tsujimoto;H. Matsui;Tetsuya Shimizu;T. Nakatsu;H. Kato;Kunihiko Watanabe
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文献类型：
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作者：
M. Akioka;H. Nakano;Aya Horikiri;Y. Tsujimoto;H. Matsui;Tetsuya Shimizu;T. Nakatsu;H. Kato;Kunihiko Watanabe
为了阐明嗜热动脉瘤杆菌AM-1菌株产生的一种特异脯氨酸氨基肽酶的结构和分子机制，克隆了其基因，在大肠杆菌中过表达重组蛋白，并用悬垂气相扩散法纯化结晶。重组氨基肽酶晶体的x射线衍射数据采集至1.8 A分辨率。晶体属于P2(1)2(1)2正交空间群，晶胞参数a = 93.62, b = 68.20, c = 76.84 a。从semet取代氨基肽酶晶体中也获得了完整的数据集。来自semet取代晶体的MAD数据集的分辨率范围为20-2.1 A的数据用于相位测定。
To elucidate the structure and molecular mechanism of a characteristic proline-specific aminopeptidase produced by the thermophile Aneurinibacillus sp. strain AM-1, its gene was cloned and the recombinant protein was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 1.8 A resolution from the recombinant aminopeptidase crystal. The crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 93.62, b = 68.20, c = 76.84 A. A complete data set was also obtained from crystals of SeMet-substituted aminopeptidase. Data in the resolution range 20-2.1 A from the MAD data set from the SeMet-substituted crystal were used for phase determination.