Targeting the biosynthesis of a novel Group A Streptococcus virulence factor

靶向新型 A 组链球菌毒力因子的生物合成

• 批准号: 2087769
• 金额: --
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2087769
• 关键词: Targeting; biosynthesis; novel; Group; Streptococcus

The Helge Dorfmueller (HD) lab is a young lab in the dynamic and vibrant Division of Molecular Microbiology. HD's research focus lies on understanding the molecular details of how the pathogen Streptococcus pyogenes (GAS) synthesises an essential virulence factor. GAS is a severe burden to life, causing a range of mild and severe, invasive infections, such as necrotising fasciitis ('flesh-eating disease') and toxic shock syndrome. It is imperative that we need to identify and characterise new drug targets to develop more efficient antibiotics and to prevent a shortfall of antibiotics in the near future. The HD lab investigates - in close collaboration with the laboratory from Prof. Mike Ferguson - the GAC biosynthesis pathway. Functional and structural characterisation will be conducted to elucidate the catalytic mechanism of this novel virulence factor, to form the basis for future antimicrobial drug development.The student will conduct fundamental and interdisciplinary research. The project and research training involves molecular microbiology and glycobiology, including cloning, protein expression and purification from recombinant bacterial system and inhibitor screening. Functional characterisation of the enzymes will be carried out using a combination of synthetic biology, bacterial genetics, carbohydrate analysis and enzyme assay development [1]. The Mike Ferguson lab provides expert training in carbohydrate purification and analysis, using biochemistry and mass-spectrometry [2].The student will be trained in inhibitor screening to identify the first chemical compounds that bind to the target enzyme. We pair glyco-enzymology with structural biology to investigate the kinetic properties of these compounds, which in combination provide the full basis for future structure-based drug design [3]. The student will have the opportunity to learn protein crystallisation and high-resolution single crystal X-ray diffraction analysis. These structural studies will reveal detailed insights into the mechanism of catalysis of these novel enzymes and the binding mode of identified inhibitors, providing the basis for future drug discovery programmes. References:[1] van der Beek, S. L., Le Breton, Y., Ferenbach, A. T., Chapman, R.N., van Aalten, D.M.F., Navratilova, I., Boons, G.J., McIver, K., van Sorge, N. M. and H.C. Dorfmueller; 'GacA is Essential for Group A Streptococcus and Defines a New Class of Monomeric dTDP-4-dehydrorhamnose Reductases (RmlD)', in print at Molecular Microbiology.[2] Allen, S., Richardson, J.M., Mehlert, A and Ferguson, M.A.J. (2013) 'Structure of a complex phosphoglycan epitope from gp72 of Trypanosoma cruzi.' J. Biol. Chem. 288, 11093-11105.[3] Dorfmueller, H.C., Borodkin, V.S., Schimpl, M., and van Aalten, D.M.F. (2009) 'GlcNAcstatins are nanomolar inhibitors of human O-GlcNAcase inducing cellular hyper-O-GlcNAcylation.' Biochem. J., 420(2), 221-7.

Helge Dorfmueller（HD）实验室是一个年轻的实验室，位于充满活力和活力的分子微生物学部门。HD的研究重点在于了解病原体化脓性链球菌（GAS）如何合成一种重要毒力因子的分子细节。GAS是一种严重的生命负担，可引起一系列轻度和重度的侵袭性感染，如坏死性筋膜炎（“食肉病”）和中毒性休克综合征。当务之急是我们需要确定和验证新的药物靶点，以开发更有效的抗生素，并防止在不久的将来出现抗生素短缺。HD实验室与Mike Ferguson教授的实验室密切合作，研究GAC生物合成途径。通过功能和结构表征来阐明这种新型毒力因子的催化机制，为未来的抗菌药物开发奠定基础。学生将进行基础和跨学科研究。该项目和研究培训涉及分子微生物学和糖生物学，包括克隆，重组细菌系统的蛋白表达和纯化以及抑制剂筛选。将结合合成生物学、细菌遗传学、碳水化合物分析和酶测定开发对酶进行功能表征[1]。Mike Ferguson实验室提供碳水化合物纯化和分析方面的专业培训，使用生物化学和质谱[2]。学生将接受抑制剂筛选的培训，以识别与目标酶结合的第一种化合物。我们将糖酶学与结构生物学结合起来研究这些化合物的动力学性质，这为未来基于结构的药物设计提供了充分的基础[3]。学生将有机会学习蛋白质结晶和高分辨率单晶X射线衍射分析。这些结构研究将揭示这些新型酶的催化机制和已识别抑制剂的结合模式的详细见解，为未来的药物发现计划提供基础。参考文献：[1]货车德比克，S。L.，Le布雷东，Y.，Ferenbach，A. T.，查普曼，R.N.，货车阿尔滕，D.M.F.，纳夫拉蒂洛娃岛Boons，G.J.，McIver，K.，货车索尔热，N. M.和H.C.之间。多夫米勒;“GacA对A组链球菌是必需的，并定义了一类新的单体dTDP-4-β-鼠李糖还原酶（RmlD β）”，在分子微生物学上发表。[2]艾伦，S.，Richardson，J.M.，Mehlert，A和Ferguson，M. A. J.（2013）“来自克氏锥虫gp 72的复合磷酸聚糖表位的结构”。288，11093-11105。[3]Dorfmueller，H.C.，Borodkin，V.S.，Schimpl，M.，和货车Aalten，D.M.F.（2009）“GlcNAc他汀是人O-GlcNAc酶的纳摩尔抑制剂，其诱导细胞高-O-GlcNAc酰化。生物化学杂志，420（2），221-7。