Plasticity of the PAS domain and a potential role for signal transduction in the histidine kinase DcuS
Plasticity of the PAS domain and a potential role for signal transduction in the histidine kinase DcuS
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DOI：
10.1038/nsmb.1493
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发表时间：
2008-10-01
影响因子：
16.8
通讯作者：
Baldus, Marc
中科院分区：
文献类型：
--
作者：
Etzkorn, Manuel;Kneuper, Holger;Baldus, Marc
目前对膜包埋传感器激酶如何识别信号和调节激酶活性的机制理解有限。在这里，我们报告的多域膜传感器激酶DcuS使用固态NMR，结构建模和诱变的结构-功能关系。将单个细胞质Per-Arnt-Sim（PAS）结构域的实验数据与计算机生成的结构模型进行比较。这些研究，连同以前的NMR工作的周质PAS域，使结构研究的膜嵌入的40 kDa的构建体的固态NMR，包括PAS段和膜域。结构改变主要限于靠近跨膜区段的蛋白质区域。来自分离的和多结构域的构建体的数据有利于在胞质结构域中的无序N-末端螺旋。该区域残基的突变强烈影响功能，表明蛋白质的灵活性与朝向激酶结构域的信号转导和激酶活性的调节有关。
The mechanistic understanding of how membrane-embedded sensor kinases recognize signals and regulate kinase activity is currently limited. Here we report structure-function relationships of the multidomain membrane sensor kinase DcuS using solid-state NMR, structural modeling and mutagenesis. Experimental data of an individual cytoplasmic Per-Arnt-Sim (PAS) domain were compared to structural models generated in silico. These studies, together with previous NMR work on the periplasmic PAS domain, enabled structural investigations of a membrane-embedded 40-kDa construct by solid-state NMR, comprising both PAS segments and the membrane domain. Structural alterations are largely limited to protein regions close to the transmembrane segment. Data from isolated and multidomain constructs favor a disordered N-terminal helix in the cytoplasmic domain. Mutations of residues in this region strongly influence function, suggesting that protein flexibility is related to signal transduction toward the kinase domain and regulation of kinase activity.