Identification of small molecules that disrupt SSB-protein interactions using a high-throughput screen

Area of Science:

• Microbiology
• Biochemistry
• Drug Discovery

Background:

• Bacterial single-stranded DNA-binding proteins (SSBs) are crucial for genome maintenance, interacting with various enzymes.
• Targeting SSB-protein interactions offers a promising strategy for developing novel antibacterial agents.
• Many of these interactions are weak, posing challenges for traditional high-throughput screening methods.

Purpose of the Study:

• To develop a high-throughput screen (HTS) for identifying small molecules that inhibit specific SSB-protein interactions.
• To target the interaction between Exonuclease I (ExoI) and the C-terminal tail of SSB (SSB-Ct).

Main Methods:

• Development of a high-throughput screening assay.
• Focus on the protein-protein interaction between Exonuclease I and the SSB C-terminal tail.
• Utilizing the binding strength between ExoI and SSB-Ct as a key assay parameter.

Main Results:

• Successfully established a high-throughput screen for identifying inhibitors of ExoI-SSB-Ct interaction.
• Demonstrated the feasibility of using this screen to find modulators of essential bacterial protein complexes.
• The screen's effectiveness relies on the inherent binding strength of the ExoI-SSB-Ct interaction.

Conclusions:

• The developed high-throughput screen is a valuable tool for discovering inhibitors of SSB-partner interactions.
• This approach can facilitate the identification of novel antibacterial compounds targeting bacterial genome maintenance pathways.
• Further research can leverage this screen to explore other weak protein-protein interactions relevant to bacterial survival.