大肠杆菌tol系统一个组分TolB的结晶及初步晶体学研究
Crystallization and preliminary crystallographic study of a component of the Escherichia coli tol system: TolB.
作者信息
Abergel C, Rigal A, Chenivesse S, Lazdunski C, Claverie J M, Bouveret E, Bénédetti H
机构信息
Information Génétique et Structurale, EP 91, Institut de Biologie Structurale et Microbiologie, 31 Chemin Joseph Aiguier, Marseille, 13402 CEDEX 20, France.
出版信息
Acta Crystallogr D Biol Crystallogr. 1998 Jan 1;54(Pt 1):102-4. doi: 10.1107/s0907444997008020.
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 P21 with unit-cell parameters a = 64.48, b = 41.06, c = 78.41 A, beta = 110.78 degrees. Frozen crystals diffract to 1.9 A 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.
来自大肠杆菌的TolB是A组大肠杆菌素用于穿透并杀死细胞的Tol系统的一部分。一个带有六个组氨酸标签的TolB衍生物被过量表达，通过在镍亲和柱上的螯合作用进行纯化，并使用SAmBA软件进行结晶以确定最佳结晶方案。晶体属于单斜晶系，空间群为P21，晶胞参数a = 64.48、b = 41.06、c = 78.41 Å，β = 110.78°。冷冻晶体的衍射分辨率为1.9 Å。目前正在对天然TolB和各种半胱氨酸取代突变体进行重原子衍生物筛选。此外，正在制备硒代甲硫氨酸取代的蛋白质，以便使用MAD方法进行结构测定。
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