Coffey Barbara M, Akhand Saeed S, Anderson Gregory G
Department of Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN 46202, USA.
Microbiology (Reading). 2014 Jun;160(Pt 6):1200-1213. doi: 10.1099/mic.0.075275-0. Epub 2014 Apr 10.
The opportunistic pathogen Pseudomonas aeruginosa causes a wide range of infections, including chronic biofilm infections in the lungs of individuals with cystic fibrosis. We previously found that the inner-membrane protein MgtE can function both as a magnesium transporter and a virulence modulator, although the exact mechanism governing these activities is unclear. To address this issue, we carried out an experimental characterization of P. aeruginosa MgtE and generated a computer-rendered model. Our in silico analysis demonstrated the structural similarity of P. aeruginosa MgtE to that of the crystal structure of MgtE in Thermus thermophilus. Experimentally, we verified that MgtE is not essential for growth and found that it may not be involved directly in biofilm formation, even under low-magnesium conditions. We demonstrated both magnesium transport and cytotoxicity-regulating functions, and showed that magnesium-binding sites in the connecting helix region of MgtE are vital in coupling these two functions. Furthermore, limiting magnesium environments stimulated mgtE transcriptional responses. Our results suggested that MgtE might play an important role in linking magnesium availability to P. aeruginosa pathogenesis.
机会致病菌铜绿假单胞菌会引发多种感染,包括囊性纤维化患者肺部的慢性生物膜感染。我们之前发现内膜蛋白MgtE既可以作为镁转运蛋白发挥作用,也可以作为毒力调节剂,尽管调控这些活性的确切机制尚不清楚。为了解决这个问题,我们对铜绿假单胞菌MgtE进行了实验表征,并生成了一个计算机渲染模型。我们的计算机模拟分析表明,铜绿假单胞菌MgtE与嗜热栖热菌中MgtE的晶体结构具有结构相似性。在实验中,我们证实MgtE对生长并非必需,并发现即使在低镁条件下,它可能也不直接参与生物膜形成。我们展示了镁转运和细胞毒性调节功能,并表明MgtE连接螺旋区域中的镁结合位点在耦合这两种功能中至关重要。此外,限制镁的环境会刺激mgtE转录反应。我们的结果表明,MgtE可能在将镁的可利用性与铜绿假单胞菌致病机制联系起来方面发挥重要作用。