基于离子淌度-质谱联用和整合建模的脲酶激活复合物结构模型。
A Structural Model of the Urease Activation Complex Derived from Ion Mobility-Mass Spectrometry and Integrative Modeling.
机构信息
Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824, USA.
出版信息
Structure. 2018 Apr 3;26(4):599-606.e3. doi: 10.1016/j.str.2018.03.001. Epub 2018 Mar 22.
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)(UreDFG) complex. Using our recently developed computational modeling workflow, we generated ensembles of putative (UreABC)(UreDFG) 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.
通过 18 个亚基酶蛋白-apoprotein-辅助蛋白前激活复合物来合成活性肺炎克雷伯氏菌脲酶在生物化学上已经得到了很好的研究，但到目前为止，这个复合物仍然难以直接进行结构特征分析。使用离子淌度-质谱法，我们对核心脲酶蛋白-apoprotein 与其辅助蛋白之间的几种蛋白质复合物进行了特征描述，包括 610 kDa 的（UreABC）（UreDFG）复合物。利用我们最近开发的计算建模工作流程，我们生成了与实验约束条件一致的假定（UreABC）（UreDFG）物种的集合，并对这些模型中的结构不确定性进行了特征描述。通过整合来自先前研究的结构信息，我们大大提高了离子淌度-质谱法衍生模型的分辨率，并且我们观察到了与该复合物所有可用数据一致的离散结构群体。
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