Monitoring Drug-Protein Interactions in the Bacterial Periplasm by Solution Nuclear Magnetic Resonance Spectroscopy

Area of Science:

• Biochemistry
• Microbiology
• Structural Biology

Background:

• Antimicrobial resistance is a global health threat, reducing treatment efficacy.
• Developing new antibiotics is crucial to combat resistant bacterial pathogens.
• Targeting bacterial periplasmic enzymes is a promising strategy for novel drug development.

Purpose of the Study:

• To present a novel in-cell Nuclear Magnetic Resonance (NMR) strategy.
• To monitor enzyme activity within the periplasmic space of Gram-negative bacteria.
• To facilitate the discovery of new drugs targeting periplasmic components.

Main Methods:

• Development of an in-cell NMR-based research strategy.
• Monitoring enzyme activity in real-time and in situ within the bacterial periplasm.
• Utilizing NMR to analyze the hydrolysis of β-lactams by β-lactamases, drug-target interactions, and inhibitor binding.

Main Results:

• Demonstrated unprecedented analytical power of in-cell NMR for periplasmic enzyme activity.
• Successfully monitored in situ and in real-time the hydrolysis of β-lactams by β-lactamases.
• Showcased the ability to track β-lactam drug interactions with targets and inhibitor binding.

Conclusions:

• In-cell NMR is a powerful tool for studying periplasmic enzymes.
• This method enables real-time monitoring of drug activity and interactions within the bacterial cell.
• The strategy supports the investigation and development of new drugs targeting the bacterial periplasm.