Cell surface β-lactamase recruitment: A facile selection to identify protein-protein interactions

Area of Science:

• Molecular Biology
• Biochemistry
• Microbiology

Background:

• Protein-protein interactions (PPIs) are fundamental to cellular functions.
• Identifying novel PPIs is crucial for discovering protein therapeutics.
• Existing methods for PPI identification can be complex or limited.

Purpose of the Study:

• To develop a simple and facile selection method for identifying protein-protein interactions.
• To link PPI-dependent protein recruitment to a selectable phenotype in bacteria.
• To provide a valuable tool for interrogating PPIs in biological research.

Main Methods:

• A selection system was engineered in Escherichia coli.
• PPIs were linked to the recruitment of β-lactamase to the bacterial cell surface.
• Bacterial survival was monitored based on ampicillin resistance.

Main Results:

• Bacteria successfully recruiting β-lactamase via PPIs were protected from ampicillin.
• Failure to recruit β-lactamase resulted in ampicillin-mediated cell death.
• The method demonstrated a PPI-dependent selectable phenotype.

Conclusions:

• A simple and tunable selection method for identifying PPIs has been established.
• This technique facilitates the discovery of novel PPIs and protein therapeutics.
• The method offers a valuable addition to the toolkit for PPI research.