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Updated: Jul 14, 2026
Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa
Published on: June 30, 2016
"Multiplexed screen identifies a Pseudomonas aeruginosa -specific small molecule targeting the outer membrane protein
Abstract:
The surge of antimicrobial resistance threatens efficacy of current antibiotics, particularly against Pseudomonas aeruginosa , a highly resistant gram-negative pathogen. The asymmetric outer membrane (OM) of P. aeruginosa combined with its array of efflux pumps provide a barrier to xenobiotic accumulation, thus making antibiotic discovery challenging. We adapted PROSPECT 1 , a target-based, whole-cell screening strategy, to discover small molecule probes that kill P. aeruginosa mutants depleted for essential proteins localized at the OM. We identified BRD1401, a small molecule that has specific activity against a P. aeruginosa mutant depleted for the essential lipoprotein, OprL. Genetic and chemical biological studies identified that BRD1401 acts by targeting the OM β-barrel protein OprH to disrupt its interaction with LPS and increase membrane fluidity. Studies with BRD1401 also revealed an interaction between OprL and OprH, directly linking the OM with peptidoglycan. Thus, a whole-cell, multiplexed screen can identify species-specific chemical probes to reveal novel pathogen biology.
Insights
Researchers discovered a new compound, BRD1401, that kills the resistant bacterium Pseudomonas aeruginosa by targeting its outer membrane. This finding offers a novel approach to combatting antimicrobial resistance.
Area of Science:
- Microbiology and Infectious Diseases
- Drug Discovery and Development
Background:
- Antimicrobial resistance (AMR) poses a significant threat, particularly against Gram-negative pathogens like Pseudomonas aeruginosa.
- The unique outer membrane (OM) and efflux pumps of P. aeruginosa present challenges for antibiotic penetration and efficacy.
Approach:
- Adapted a target-based, whole-cell screening strategy (PROSPECT) to identify compounds targeting OM proteins.
- Screened for small molecules that kill P. aeruginosa mutants lacking essential OM proteins.
Key Points:
- Identified BRD1401, a novel small molecule with specific activity against a P. aeruginosa mutant lacking the essential lipoprotein OprL.
- BRD1401 targets the OM beta-barrel protein OprH, disrupting its interaction with lipopolysaccharide (LPS) and increasing membrane fluidity.
- Revealed a direct link between OprL and OprH, connecting the OM to the peptidoglycan layer.
Conclusions:
- Whole-cell, multiplexed screening can identify species-specific chemical probes.
- This approach can uncover novel pathogen biology and lead to new strategies against resistant bacteria like P. aeruginosa.
