research
Chirality: The Key to Specific Bacterial Protease-Based Diagnosis?
- Publication date
- 24 January 2014
- Publisher
- Abstract
Bacterial proteases play an important role in a broad spectrum of processes, including
colonization, proliferation and virulence. In this respect, bacterial proteases are
potential biomarkers for bacterial diagnosis and targets for novel therapeutic protease
inhibitors. To investigate these potential functions, the authors designed and used a
protease substrate fluorescence resonance energy transfer (FRET)-library comprising
115 short D- and L-amino-acid-containing fluorogenic substrates as a tool to generate
proteolytic profiles for a wide range of bacteria. Bacterial specificity of the D-amino acid
substrates was confirmed using enzymes isolated from both eukaryotic and prokaryotic
organisms. Interestingly, bacterial proteases that are known to be involved in housekeeping
and nutrition, but not in virulence, were able to degrade substrates in which a
D-amino acid was present. Using our FRET peptide library and culture supernatants from
a total of 60 different bacterial species revealed novel, bacteria-specific, proteolytic
profiles. Although in-species variation was observed for Pseudomonas aeruginosa, Porphyromonas
gingivalis and Staphylococcus aureus. Overall, the specific characteristic of
our substrate peptide library makes it a rapid tool to high-throughput screen for novel
substrates to detect bacterial proteolytic activity.