Year: 2014
Type: dissertation
Abstract: \n \nBacterial proteases play an important role in a broad spectrum of processes, including \ncolonization, proliferation and virulence. In this respect, bacterial proteases are \npotential biomarkers for bacterial diagnosis and targets for novel therapeutic protease \ninhibitors. To investigate these potential functions, the authors designed and used a \nprotease substrate fluorescence resonance energy transfer (FRET)-library comprising \n115 short D- and L-amino-acid-containing fluorogenic substrates as a tool to generate \nproteolytic profiles for a wide range of bacteria. Bacterial specificity of the D-amino acid \nsubstrates was confirmed using enzymes isolated from both eukaryotic and prokaryotic \norganisms. Interestingly, bacterial proteases that are known to be involved in housekeeping \nand nutrition, but not in virulence, were able to degrade substrates in which a \nD-amino acid was present. Using our FRET peptide library and culture supernatants from \na total of 60 different bacterial species revealed novel, bacteria-specific, proteolytic \nprofiles. Although in-species variation was observed for Pseudomonas aeruginosa, Porphyromonas \ngingivalis and Staphylococcus aureus. Overall, the specific characteristic of \nour substrate peptide library makes it a rapid tool to high-throughput screen for novel \nsubstrates to detect bacterial proteolytic activity.
Author: Wendy E. Kaman
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2170456649