Department of Chemistry, Imperial College London, UK.
ACS Chem Biol. 2010 Mar 19;5(3):279-85. doi: 10.1021/cb9002859.
Clostridium difficile, a leading cause of hospital-acquired infection, possesses a dense surface layer (S-layer) that mediates host-pathogen interactions. The key structural components of the S-layer result from proteolytic cleavage of a precursor protein, SlpA, into high- and low-molecular-weight components. Here we report the discovery and optimization of the first inhibitors of this process in live bacteria and their application for probing S-layer processing. We also describe the design and in vivo application of activity-based probes that identify the protein Cwp84 as the cysteine protease that mediates SlpA cleavage. This work provides novel chemical tools for the analysis of S-layer biogenesis and for the potential identification of novel drug targets within clostridia and related bacterial pathogens.
艰难梭菌是医院获得性感染的主要原因，它具有密集的表面层（S 层），介导宿主-病原体相互作用。S 层的关键结构成分是由前体蛋白 SlpA 经蛋白水解切割成高分子量和低分子量成分产生的。在这里，我们报告了在活细菌中首次发现和优化这种过程的抑制剂，并将其应用于探测 S 层加工。我们还描述了活性探针的设计和体内应用，这些探针确定了半胱氨酸蛋白酶 Cwp84 作为介导 SlpA 切割的蛋白酶。这项工作为 S 层生物发生的分析以及在梭菌和相关细菌病原体中潜在的新型药物靶点的鉴定提供了新的化学工具。
