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

该项目将为博士生提供一个极好的机会，可以跨多个学科进行培训，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。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拓扑结构，这是一个迄今为止仍然难以解决的问题。结合谢菲尔德大学最新的单分子成像技术，这项工作有望推动科学发现的界限，并导致该领域的重大突破。博士生将受益于来自不同学科的两位导师提供的刺激性跨学科专业知识（S. Mesnage是一位微生物遗传学家，在细胞壁生物化学方面有很强的背景。威廉姆森是蛋白质结构方面的生物制药专家）。成功的候选人将接受几种互补方法的培训：（一）蛋白质工程的分子生物学和生物化学;（二）细菌细胞壁分析的HPLC和质谱法;（iii）表面等离子体共振，微量热（差示扫描量热法和等温滴定量热法）和NMR用于蛋白质-PG相互作用，及（iv）超分辨率荧光显微镜研究蛋白质的亚细胞定位。博士生的培训也将涉及与Simon Jones博士团队的密切互动（化学系，谢菲尔德大学），他目前正在开发生产肽聚糖片段的合成方案。

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.