title: Chemical probes of surface layer biogenesis in Clostridium difficile.
abstract: 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.
authors.affiliation: Department of Chemistry, Imperial College London, UK.
authors.forename: T H Tam
authors.initials: TH
authors.lastname: Dang
authors.forename: Lucia
authors.initials: L
authors.lastname: de la Riva
authors.forename: Robert P
authors.initials: RP
authors.lastname: Fagan
authors.forename: Elisabeth M
authors.initials: EM
authors.lastname: Storck
authors.forename: William P
authors.initials: WP
authors.lastname: Heal
authors.forename: Claire
authors.initials: C
authors.lastname: Janoir
authors.forename: Neil F
authors.initials: NF
authors.lastname: Fairweather
authors.forename: Edward W
authors.initials: EW
authors.lastname: Tate
mesh_terms: D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D016360:Clostridioides difficile / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans
chemical_list: D001426:Bacterial Proteins; C070683:surface layer protein A, Bacteria; C524258:Cwp84 protein, Clostridium difficile; D003546:Cysteine Endopeptidases