Crystallization and preliminary crystallographic study of a component of the Escherichia coli Tol system: TolB
Crystallization and preliminary crystallographic study of a component of the Escherichia coli Tol system: TolB
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DOI：
10.1107/s0907444997008020
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发表时间：
1998-01-01
影响因子：
2.2
通讯作者：
Benedetti, H
中科院分区：
文献类型：
--
作者：
Abergel, C;Rigal, A;Benedetti, H
来自大肠杆菌的 TolB 是 A 族大肠杆菌素用来穿透和杀死细胞的 Tol 系统的一部分。用六个组氨酸标记的 TolB 衍生物被过表达，通过镍亲和柱上的螯合纯化，并使用 SAmBA 软件进行结晶，以确定最佳结晶方案。该晶体属于单斜晶系，空间群 P2(1)，晶胞参数 a = 64.48，b = 41.06，c = 78.41 埃，β = 110.78 度。冷冻晶体的衍射分辨率为 1.9 埃。对天然 TolB 和各种半胱氨酸取代突变体的重原子衍生物的筛选正在进行中。此外，为了使用 MAD 方法进行结构测定，正在生产一种硒代蛋氨酸取代的蛋白质。
TolB from Escherichia coli is part of the Tol system used by the group A colicins to penetrate and kill cells. A TolB derivative tagged with six histidines was overexpressed, purified by chelation on a nickel affinity column and crystallized using the SAmBA software to define the optimal crystallization protocol. The crystals belong to the monoclinic system, space group P2(1) with unit-cell parameters a = 64.48, b = 41.06, c = 78.41 Angstrom, beta = 110.78 degrees. Frozen crystals diffract to 1.9 Angstrom resolution. Screening for heavy-atom derivatives both on the native TolB and various cysteine-substituted mutants is in progress. In addition, a selenomethionine-substituted protein is being produced in order to use the MAD method for structure determination.