A Structural Model of the Urease Activation Complex Derived from Ion Mobility-Mass Spectrometry and Integrative Modeling
A Structural Model of the Urease Activation Complex Derived from Ion Mobility-Mass Spectrometry and Integrative Modeling
复制标题
DOI：
10.1016/j.str.2018.03.001
复制
发表时间：
2018-04-03
期刊：
影响因子：
5.7
通讯作者：
Ruotolo, Brandon T.
中科院分区：
文献类型：
--
作者：
Eschweiler, Joseph D.;Farrugia, Mark. A.;Ruotolo, Brandon T.
通过18亚基酶辅基蛋白-辅助蛋白预活化复合体合成活性产气克雷伯菌尿素酶已被广泛的生化研究，但到目前为止，该复合体仍难以直接进行结构表征。利用离子迁移率-质谱法，我们鉴定了核心尿素酶脱辅基蛋白与其辅助蛋白之间的几种蛋白质复合体，包括610 kDa(UreABC)(3)(UreDFG)(3)复合体。使用我们最近开发的计算建模工作流，我们生成了与实验限制一致的假定(UreABC)(3)(UreDFG)(3)物种的系综，并表征了这些模型中存在的结构模糊性。通过整合以前研究的结构信息，我们大大提高了离子迁移率-质谱学衍生模型的分辨率，我们观察到与该复合体的所有可用数据一致的离散结构群体。
The synthesis of active Klebsiella aerogenes urease via an 18-subunit enzyme apoprotein-accessory protein pre-activation complex has been well studied biochemically, but thus far this complex has remained refractory to direct structural characterization. Using ion mobility-mass spectrometry, we characterized several protein complexes between the core urease apoprotein and its accessory proteins, including the 610-kDa (UreABC)(3)(UreDFG)(3) complex. Using our recently developed computational modeling workflow, we generated ensembles of putative (UreABC)(3)(UreDFG)(3) species consistent with experimental restraints and characterized the structural ambiguity present in these models. By integrating structural information from previous studies, we increased the resolution of the ion mobility-mass spectrometry-derived models substantially, and we observe a discrete population of structures consistent with all of the available data for this complex.