El-Halfawy Omar M, Brown Eric D
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
Michael G. DeGroote Institute of Infectious Disease Research, McMaster University, Hamilton, ON, Canada.
Methods Mol Biol. 2019;1954:297-308. doi: 10.1007/978-1-4939-9154-9_23.
The world is heading toward a dangerous post-antibiotic era where antibiotics fail to treat infections. Staphylococcus aureus is the leading cause of healthcare-associated infections worldwide, and an ever-increasing percentage of them are methicillin-resistant (MRSA). New strategies are urgently needed to combat this pathogen. Wall teichoic acids (WTA) in S. aureus are polyribitol phosphate polymers that play important roles in virulence and resistance to β-lactam antibiotics. Here, we describe a high-throughput whole-cell screening platform for inhibitors targeting WTA biosynthesis. This platform takes advantage of the unique dispensability patterns of genes encoding WTA biosynthesis. We further describe follow-up dose-response assays to identify WTA inhibitors among the primary bioactives. WTA inhibitors offer an exciting opportunity for the development of novel antibacterial leads of unique mechanism in the fight against drug-resistant staphylococcal infections.
世界正朝着一个危险的后抗生素时代迈进，在这个时代，抗生素无法治疗感染。金黄色葡萄球菌是全球医疗保健相关感染的主要原因，其中耐甲氧西林（MRSA）的比例不断增加。迫切需要新的策略来对抗这种病原体。金黄色葡萄球菌中的壁磷壁酸（WTA）是聚核糖醇磷酸聚合物，在毒力和对β-内酰胺抗生素的抗性中起重要作用。在这里，我们描述了一个针对靶向WTA生物合成的抑制剂的高通量全细胞筛选平台。该平台利用了编码WTA生物合成的基因独特的可 dispensability 模式。我们进一步描述了后续的剂量反应测定，以在主要生物活性物质中鉴定WTA抑制剂。WTA抑制剂为开发具有独特机制的新型抗菌先导物提供了一个令人兴奋的机会，以对抗耐药性葡萄球菌感染。
