Activity-Based Protein Profiling Identifies an α-Amylase Family Protein Contributing to the Virulence of...

Area of Science:

• Microbiology
• Biochemistry
• Drug Discovery

Background:

• Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant public health threat, necessitating novel antimicrobial strategies.
• Identifying essential bacterial enzymes can reveal new drug targets for combating antibiotic resistance.

Purpose of the Study:

• To identify and characterize retaining glycosidase activities in MRSA as potential antimicrobial drug targets.
• To investigate the roles of novel identified glycosidases, specifically BglA and TreC, in bacterial physiology and virulence.

Main Methods:

• Activity-based protein profiling (ABPP) using fluorescent and biotinylated probes to detect and enrich glycosidase activities.
• Mass spectrometry for identification of enriched glycoside hydrolase family proteins.
• CRISPR interference (CRISPRi) and inhibitor studies to assess the physiological relevance of identified enzymes.

Main Results:

• Three glycoside hydrolase family proteins were identified: 6-phospho-β-glucosidase (BglA), trehalase C (TreC), and autolysin (Atl).
• Silencing of treC in MRSA reduced in vitro biofilm formation and attenuated virulence in a Galleria mellonella infection model.
• BglA and TreC were identified as previously uncharacterized enzymes with potential roles in MRSA pathogenesis.

Conclusions:

• Retaining glycosidases represent a promising area for discovering new antimicrobial drug targets against MRSA.
• TreC warrants further investigation for its biochemical function and potential as a therapeutic target due to its impact on biofilm formation and virulence.