Investigating the mechanism of phosphate aquisition by Pseudomonas aeruginosa

铜绿假单胞菌获取磷酸盐的机制研究

• 批准号: 401975-2011
• 负责人: Moraes, Trevor
• 金额: $ 2.99万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=573117
• 关键词: Investigating; mechanism; phosphate; aquisition; Pseudomonas

Phosphate is an essential nutrient that is found in lipids, nucleic acids, proteins and sugars and is also key to many biological reactions that require ATP and/or signaling pathways that exploit protein phosphorylation. Thus, bacteria utilize a global regulatory circuit, the Pho regulon, to manage intracellular bacterial phosphate homeostasis. The Pho regulon regulates the expression of key proteins involved in the acquisition of phosphate, including integral membrane proteins that play an important role in the transport of phosphate across lipid bilayers and into the cell. These membrane proteins also act as sensory proteins that interact with phosphate binding proteins to detect the levels of extracellular phosphate and in turn regulate their own expression through the Pho regulon. The Pho regulon pathway also controls many other gene networks including the expression and secretion of virulence factors in response to phosphate levels, thus this phosphate acquisition pathway in bacteria is extremely important. The integral membrane proteins (OprP and PstABC) that translocate phosphate interact with their aqueous soluble transport partners (PstS and PhoU) facilitating the transport of these anions and also initiate signaling events that are propagated to control gene expression. Research in the Moraes lab will examine the protein interactions between integral membrane protein transporters and their soluble transporter counterparts that lead to transport and signal transduction. We plan to dissect the components of this pathway and their interactions in molecular detail using tools such as X-ray crystallography to provide atomic resolution 3D- images of the proteins involved in phosphate translocation. Indeed, the high-resolution structure of the outer membrane protein OprP determined by Dr. Moraes provides insight into the first step of phosphate acquisition. Highly qualified personnel will be well trained in the areas of membrane protein structural biology, microbiology, biochemistry and molecular biology, preparing them for careers in basic scientific research and biotechnology.

磷酸盐是一种必需的养分，它在脂质，核酸，蛋白质和糖中发现，也是许多需要ATP和/或信号传导途径的生物反应的关键，这些途径可以利用蛋白质磷酸化。 因此，细菌利用全球调节回路PHO调节量来管理细胞内细菌磷酸盐稳态。 PHO调节量调节与磷酸盐采集有关的关键蛋白的表达，包括整体膜蛋白，这些膜蛋白在磷酸盐跨脂质双层和细胞中发挥重要作用。 这些膜蛋白还充当感官蛋白，与磷酸盐结合蛋白相互作用以检测细胞外磷酸盐的水平，进而通过Pho congulon调节其自身的表达。 PHO调节途径还控制着许多其他基因网络，包括响应磷酸盐水平的毒力因子的表达和分泌，因此细菌中这种磷酸盐采集途径非常重要。 磷酸易位的磷酸盐的整合膜蛋白（OPRP和PSTABC）与它们的水溶性可溶性传输伙伴（PSTS和PHOU）相互作用，促进了这些阴离子的传输，还启动了传播以控制基因表达的信号传导事件。 MORAES实验室中的研究将检查积分膜蛋白转运蛋白与其可溶性转运蛋白对应物之间的蛋白质相互作用，从而导致运输和信号转导。我们计划使用诸如X射线晶体学等工具，以提供参与磷酸盐易位的蛋白质的原子分辨率3D图像，以分子细节进行分子细节的分子相互作用。实际上，由Moraes博士确定的外膜OPRP的高分辨率结构提供了对磷酸盐采集的第一步的见解。高素质的人员将在膜蛋白结构生物学，微生物学，生物化学和分子生物学领域接受良好的培训，为基础科学研究和生物技术的职业做好准备。