In Situ Biofilm Affinity-Based Protein Profiling Identifies the Streptococcal Hydrolase GbpB as the Target of a Carolacton-Inspired Chemical Probe

J Am Chem Soc. 2024 Aug 21;146(33):23449-23456. doi: 10.1021/jacs.4c06658. Epub 2024 Aug 12.

Abstract

Natural products are important precursors for antibiotic drug design. These chemical scaffolds serve as synthetic inspiration for chemists who leverage their structures to develop novel antibacterials and chemical probes. We have previously studied carolacton, a natural product macrolactone fromSorangium cellulosum, and discovered a simplified derivative, A2, that maintained apparent biofilm inhibitory activity, although the biological target was unknown. Herein, we utilize affinity-based protein profiling (AfBPP) in situ during biofilm formation to identify the protein target using a photoexcitable cross-linking derivative of A2. From these studies, we identified glucan binding protein B (GbpB), a peptidoglycan hydrolase, as the primary target of A2. Further characterization of the interaction between A2 and GbpB, as well as PcsB, a closely related homologue from the more pathogenic S. pneumoniae, revealed binding to the catalytic CHAP (cysteine, histidine, aminopeptidase) domain. To the best of our knowledge, this is the first report of a small-molecule binder of a conserved and essential bacterial CHAP hydrolase, revealing its potential as an antibiotic target. This work also highlights A2 as a useful tool compound for streptococci and as an initial scaffold for the design of more potent CHAP binders.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Biofilms* / drug effects
  • Hydrolases / antagonists & inhibitors
  • Hydrolases / chemistry
  • Hydrolases / metabolism
  • Lactones / chemistry
  • Lactones / metabolism
  • Lactones / pharmacology
  • Molecular Probes / chemistry
  • Molecular Probes / metabolism
  • N-Acetylmuramoyl-L-alanine Amidase / chemistry
  • N-Acetylmuramoyl-L-alanine Amidase / metabolism

Substances

  • Bacterial Proteins
  • Anti-Bacterial Agents
  • Molecular Probes
  • N-Acetylmuramoyl-L-alanine Amidase
  • Lactones
  • Hydrolases