Crystallization and preliminary X-ray diffraction analysis of the full-length Mg2+ transporter MgtE
Crystallization and preliminary X-ray diffraction analysis of the full-length Mg2+ transporter MgtE
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DOI：
10.1107/s1744309107032332
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发表时间：
2007-08-01
影响因子：
0.9
通讯作者：
Nureki, Osamu
中科院分区：
文献类型：
--
作者：
Hattori, Motoyuki;Tanaka, Yoshiki;Nureki, Osamu
Mg2+转运蛋白的MgtE家族在所有三个结构域中都普遍保守。从7个不同物种中克隆了编码全长MgtE的基因。七种MgtE转运蛋白中的三种被过表达并纯化用于结晶试验。只有嗜热栖热菌MgtE成功地结晶使用坐滴蒸气扩散法。硒蛋氨酸取代（SeMet）晶体，获得交叉微晶种使用本机微晶。SeMet晶体使用同步辐射衍射X射线至3.5 A分辨率，并且属于空间群C222（1），具有晶胞参数a = 118.3，B = 134.9，c = 366.2 A。结构测定正在进行中。
The MgtE family of Mg2+ transporters are ubiquitously conserved in all three domains. The genes encoding full-length MgtE from seven different species were cloned. Three of the seven MgtE transporters were overexpressed and purified for use in crystallization trials. Only Thermus thermophilus MgtE was successfully crystallized using the sitting-drop vapour-diffusion method. Selenomethionine-substituted (SeMet) crystals were obtained by cross-micro-seeding using the native microcrystals. The SeMet crystals diffracted X-rays to 3.5 A resolution using synchrotron radiation and belong to space group C222(1), with unit-cell parameters a = 118.3, b = 134.9, c = 366.2 A. Structure determination is in progress.