Understanding the molecular mechanism of bacterial sphingolipid synthesis

细菌产生广泛的膜脂，我们对这些膜脂的了解正在增长，最近的工作表明，它们产生的脂类以前被认为只由真核生物产生，包括一系列不同的鞘磷脂。鞘磷脂是真核细胞膜的关键结构成分，在细胞内和细胞间信号转导中也起着关键作用。最近在关键的人类微生物组物种中发现了它们，包括拟杆菌和大肠杆菌。该项目的目的是确定在大肠杆菌中神经鞘脂脂生物合成早期阶段涉及的关键酶的结构和功能，并了解这些酶的表达是如何响应环境和宿主因素的。在这个项目中，学生将使用分子克隆来产生蛋白质表达构建体；他们将在大肠杆菌表达宿主中产生重组蛋白质；并使用层析方法来纯化重组蛋白质。学生将发展结构生物学和生物化学方法的技能，以表征这些蛋白质的结构和功能。这个多学科项目将结构生物学和生物化学的专业知识与关键宿主/微生物相互作用中的基因调控网络研究的专业知识结合起来，结构生物学方面的其他专业知识将来自Durham Partners。学生还将有广泛的机会与我们在爱丁堡大学和罗格斯大学的合作者一起工作。最终，这个项目将帮助我们了解细菌用来产生这些重要膜脂的生物合成途径，以及它们的表达是如何在宿主环境中受到控制的。该项目的成果将对了解宿主微生物的相互作用产生影响，并可能为抗菌分子的开发提供新的靶点。

Bacteria produce a wide range of membrane lipids and our understanding of these is growing, with recent work showing that they produce lipids that were previously thought to only be produced by eukaryotes, including an array of different sphingolipids. Sphingolipids are key structural components of eukaryotic membranes, and they also play key roles in intra- and inter-cellular signalling. Recently they have been discovered in key human microbiome species, including Bacteroides thetaiotaomicron and Escherichia coli.The project aims are to determine the structure and function of the key enzymes involved in the early stages of sphingolipid biosynthesis in E. coli and understand how the expression of these enzymes is regulated in response to environmental and host factors. In this project the student will use molecular cloning to produce protein expression constructs; they will produce recombinant protein in an Escherichia coli expression host; and use chromatography methods to purify the recombinant proteins. The student will develop skills in structural biology and biochemistry methods to characterise the structure and function of these proteins. This multidisciplinary project combines expertise in structural biology and biochemistry with expertise in the study of gene regulatory networks in key host/microbe interactions additional expertise in structural biology will come from Durham partner. The student will also have extensive opportunities to work with our collaborators at Edinburgh University and Rutgers University. Ultimately this project will help us develop our understanding of the biosynthetic pathways used by bacteria to produce these important membrane lipids and how their expression is controlled in a host context. The project outcomes will have impact in understanding host microbe interactions and may provide new targets for the development of antimicrobial molecules.