Crystallization and preliminary X-ray diffraction analysis of a novel-type phosphoserine phosphatase from Hyd rogenobac t e r the rmoph i1 us TK-6.
Crystallization and preliminary X-ray diffraction analysis of a novel-type phosphoserine phosphatase from Hyd rogenobac t e r the rmoph i1 us TK-6.
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来自氢杆菌 TK-6 的新型磷酸丝氨酸磷酸酶的结晶和初步 X 射线衍射分析。
DOI：
10.1107/s1744309112025213
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
M. (*Equal contribution)
中科院分区：
文献类型：
--
作者：
Chiba;Y.;* Horita;S.;* Ohtsuka;J.;Arai;H.;Nagata;K.;Igarashi;Y.;Tanokura;M. and Ishii;M. (*Equal contribution)
从嗜热和氧化氢的嗜热氢杆菌TK-6中鉴定出两种具有独特底物特异性的新型磷酸丝氨酸磷酸酶（PSPs）。其中一种PSPs （iPSP1）在大肠杆菌中异种表达，纯化并结晶。以PEG 4000为沉淀剂，采用坐滴气相扩散法获得了衍射质量的晶体。从单晶中采集了分辨率分别为45.0-2.50和45.0-1.50 Å的两个衍射数据集，并进行合并，得到了一个高度完整的数据集。该晶体的空间群为原始正交P212121，晶胞参数a = 49.8, b = 73.6, c = 124.3 Å。计算的Matthews系数（VM = 2.32 Å3 Da−1）表明该晶体每个不对称单元含有一个iPSP1配合物。
Two novel-type phosphoserine phosphatases (PSPs) with unique substrate specificity from the thermophilic and hydrogen-oxidizing bacterium Hydrogenobacter thermophilus TK-6 have previously been identified. Here, one of the PSPs (iPSP1) was heterologously expressed in Escherichia coli, purified and crystallized. Diffraction-quality crystals were obtained by the sitting-drop vapour-diffusion method using PEG 4000 as the precipitant. Two diffraction data sets with resolution ranges of 45.0–2.50 and 45.0–1.50 Å were collected from a single crystal and were merged to give a highly complete data set. The space group of the crystal was identified as primitive orthorhombic P212121, with unit-cell parameters a = 49.8, b = 73.6, c = 124.3 Å. The calculated Matthews coefficient (VM = 2.32 Å3 Da−1) indicated that the crystal contained one iPSP1 complex per asymmetric unit.