Cell surface β-lactamase recruitment: A facile selection to identify protein-protein interactions.
Jordan A HinmonJade M KingLatrina J MayoCierra R FariesYa'hnis T LockettDavid W CrawfordPatrick C BeardsleeAlexander HendricksBrian R McNaughtonPublished in: Protein science : a publication of the Protein Society (2024)
Protein-protein interactions (PPIs) are central to many cellular processes, and the identification of novel PPIs is a critical step in the discovery of protein therapeutics. Simple methods to identify naturally existing or laboratory evolved PPIs are therefore valuable research tools. We have developed a facile selection that links PPI-dependent β-lactamase recruitment on the surface of Escherichia coli with resistance to ampicillin. Bacteria displaying a protein that forms a complex with a specific protein-β-lactamase fusion are protected from ampicillin-dependent cell death. In contrast, bacteria that do not recruit β-lactamase to the cell surface are killed by ampicillin. Given its simplicity and tunability, we anticipate this selection will be a valuable addition to the palette of methods for illuminating and interrogating PPIs.
Keyphrases
- escherichia coli
- cell surface
- klebsiella pneumoniae
- protein protein
- cell death
- small molecule
- multidrug resistant
- gram negative
- amino acid
- binding protein
- magnetic resonance
- quantum dots
- computed tomography
- biofilm formation
- reduced graphene oxide
- high throughput
- gold nanoparticles
- staphylococcus aureus
- metal organic framework
- cell proliferation
- signaling pathway
- single cell
- visible light
- pi k akt