Functional characterization of a novel peptidoglycan binding domain conserved in Gram-positive nosocomial pathogens

对于博士生来说，这个项目将是一个极好的机会，可以接受包括分子生物学、生物化学、生物物理学、结构生物学和细胞生物学在内的多个学科的培训。它还将采用谢菲尔德大学开发的最先进的超分辨率显微镜技术。在这项工作的背景下开发的新型荧光探针将能够探索肽聚糖的拓扑结构，这是一个到目前为止仍然难以解决的问题。与谢菲尔德大学最新的单分子成像技术相结合，这项工作有望突破科学发现的界限，在该领域取得重大突破。博士生将受益于来自不同学科的两位导师提供的富有启发性的跨学科专业知识(S.Mesnage是具有丰富细胞壁生物化学背景的微生物遗传学家；M.Williamson是蛋白质结构方面的生物物理学家专家)。成功的候选人将接受几种辅助方法的培训：(I)蛋白质工程的分子生物学和生物化学；(Ii)用于细菌细胞壁分析的高效液相色谱和质谱学；(Iii)用于蛋白质-PG相互作用的表面等离子体共振、微热量(差示扫描量热法和等温滴定热法)和核磁共振；以及(Iv)用于研究蛋白质的亚细胞定位的超分辨率荧光显微镜。博士生的培训还将涉及与谢菲尔德大学化学系的Simon Jones博士团队的密切互动，后者目前正在开发合成方案以生产肽聚糖片段。

This project will represent a fantastic opportunity for a PhD student to have training across several disciplines including molecular biology, biochemistry, biophysics, structural biology and cell biology. It will also embrace state-of-the-art super resolution microscopy technologies developed at the University of Sheffield. The novel fluorescent probes developed in the context of this work will enable to explore peptidoglycan topology, a question that has remained intractable so far. Combined with the latest single-molecule imaging technologies available at the university of Sheffield, this work is therefore expecting to push the boundaries of scientific discovery and lead to major breakthrough in the field.The PhD student will benefit from a stimulating interdisciplinary expertise provided by two supervisors from different disciplines (S. Mesnage is a microbial geneticist with strong background in cell wall biochemistry; M. Williamson is a biophysicist expert in protein structure). The successful candidate will be trained in several complementary approaches: (i) molecular biology and biochemistry for protein engineering; (ii) HPLC and mass spectrometry for bacterial cell wall analysis; (iii) Surface Plasmon Resonance, microcalorimety (Differential Scanning Calorimetry and Isothermal Titration Calorimetry) and NMR for protein-PG interactions, and (iv) super-resolution fluorescence microscopy to study the subcellular localization of proteins.The training of the PhD student will also involve a close interaction with the team of Dr Simon Jones (department of Chemistry, University of Sheffield), who is currently developing synthetic schemes to produce peptidoglycan fragments.