Department of Environmental Protection, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Granada, Spain.
Department of Physical Chemistry, Faculty of Chemistry, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30071 Murcia, Spain.
Sci Rep. 2016 Jun 29;6:28967. doi: 10.1038/srep28967.
Inorganic phosphate (Pi) is a central signaling molecule that modulates virulence in various pathogens. In Pseudomonas aeruginosa, low Pi concentrations induce transcriptional alterations that increase virulence. Also, under low Pi levels, P. aeruginosa exhibits Pi chemotaxis-a process mediated by the two non-paralogous receptors CtpH and CtpL. Here we show that the two receptors operate via different mechanisms. We demonstrate that the ligand binding domain (LBD) of CtpH but not CtpL binds Pi directly. We identify the periplasmic ligand binding protein PstS as the protein that binds in its Pi loaded state to CtpL, resulting in receptor stimulation. PstS forms part of the Pi transporter and has thus a double function in Pi transport and chemotaxis. The affinity of Pi for CtpH was modest whereas that for PstS very high, which may explain why CtpH and CtpL mediate chemotaxis to high and low Pi concentrations, respectively. The pstS/ctpH double mutant was almost devoid of Pi taxis, indicating that PstS is the only CtpL Pi-shuttle. Chemotaxis mechanisms based on indirect ligand recognition were unambiguously identified in enterobacteria. The discovery of a similar mechanism in a different bacterial order, involving a different chemoreceptor type and chemoeffector suggests that such systems are widespread.
无机磷酸盐(Pi)是一种重要的信号分子,可调节多种病原体的毒力。在铜绿假单胞菌中,低 Pi 浓度会诱导转录改变,从而增加毒力。此外,在低 Pi 水平下,铜绿假单胞菌表现出 Pi 趋化性,这是由两个非同源受体 CtpH 和 CtpL 介导的过程。在这里,我们表明这两个受体通过不同的机制起作用。我们证明 CtpH 的配体结合域(LBD)而不是 CtpL 可以直接结合 Pi。我们确定周质配体结合蛋白 PstS 是在其 Pi 加载状态下与 CtpL 结合的蛋白,从而导致受体刺激。PstS 是 Pi 转运体的一部分,因此在 Pi 转运和趋化性中具有双重功能。Pi 与 CtpH 的亲和力适中,而与 PstS 的亲和力非常高,这可能解释了为什么 CtpH 和 CtpL 分别介导对高浓度和低浓度 Pi 的趋化性。pstS/ctpH 双突变体几乎没有 Pi 趋化性,表明 PstS 是 CtpL 唯一的 Pi 穿梭蛋白。基于间接配体识别的趋化作用机制在肠杆菌中得到了明确的鉴定。在不同的细菌目中发现了类似的机制,涉及不同的化学感受器类型和化学效应物,表明这种系统广泛存在。