Chemisch-proteomische Strategien zur Identifikation krankheitsassoziierter Enzyme in pathogenen Bakterien als neuartige Angriffsziele für Antibiotika
Final Report Abstract
After decades of successful treatment of bacterial infections with antibiotics, formerly treatable bacteria have developed drug resistance and consequently pose a major threat to public health. To address the urgent need for effective antibacterial drugs we developed a streamlined chemical-biology platform that facilitates the consolidated identification and structural elucidation of natural products together with their dedicated cellular targets. To identify novel targets for the treatment of multidrug resistant S. aureus (MRSA) strains, we recently introduced functionalized small biomimetic β-lactone, β-lactam, Michael acceptor and maleimide natural product derived molecules. These compounds were modified with a small tag for the visualization and identification of dedicated targets in complex proteomes by SDS- gel-electrophoresis and mass spectrometry. Structural variations in side chains of selected molecules led to an increased affinity for certain enzymes that played crucial roles in resistance and virulence. The general utility of this approach was demonstrated by the chemical knockout of a central S. aureus virulence regulator, ClpP, that resulted in a drastically decreased expression of major virulence factors which are key players in e.g. sepsis, tissue necrosis, inflammation and toxic shock. Since ClpP is highly conserved in many pathogens, this strategy could represent a global approach for the treatment of infectious diseases by disarming the bacterial virulence repertoire. In fact, follow up studies with Listeria monocytogenes and Plasmodium falciparum, the causative pathogen of malaria, revealed anti-virulence activities of the compound also for these highly different organisms. We received for these results the “Innovation Award of the German Bioregions” in 2008 as well as the “Arnold Sommerfeld Award” of the Bavarian Acadamy of Sciences in 2009. The innovation was patented and will be soon transferred into an independent spin off. In addition to these results we introduced diverse chemical tools to monitor bacterial resistance with small molecules that revealed new and previously uncharacterized resistance associated enzymes in multiresistant bacteria as well as new strategies for the selective release of captured biomolecules from solid support.
Publications
- “β-Lactams as selective chemical probes for the in vivo labeling of bacterial enzymes involved in cell wall biosynthesis, antibiotic resistance and virulence” J. Am. Chem. Soc., 2008, 130, 13400-13407
Staub, I., Sieber, S. A.
- “β-Lactones as privileged structures for the active site labeling of versatile bacterial enzyme classes” Angewandte Chemie, 2008, 47, 4600- 46003
Böttcher, T., Sieber, S. A.
- “β-Lactones as specific inhibitors of ClpP attenuate the production of extracellular virulence factors of Staphylococcus aureus” J. Am. Chem. Soc., 2008, 130, 14400-14401
Böttcher, T., Sieber, S. A.
- "β -Lactam probes as selective chemical-proteomic tools for the identification and functional characterization of resistance associated enzymes in MRSA" J. Am. Chem. Soc., 2009, 131, 6271-6276
Staub, I., and Sieber, S. A.
- beta-Lactones reduce the intracellular virulence of Listeria monocytogenes in macrophages” ChemMedChem, 2009, 4, 1260-1263
Böttcher, T., and Sieber, S. A.
- “A new generation of structurally refined β-lactones as potent inhibitors of devastating bacterial virulence factors” ChemBioChem, 2009, 10, 663-666
Böttcher, T., Sieber, S. A.
- “A photolabile linker for the mild and selective cleavage of enriched biomolecules from solid support” J. Org. Chem., 2009, 74, 8476-8479
Orth, R., Sieber, S. A.
- “Cinnamic aldehyde derived probes for the active site labelling of pathogenesis associated enzymes” Chem. Commun, 2009, 3741-3743
Pitscheider, M., Sieber, S. A.
- “Unravelling the secrets of Protein-metabolite interactions“ ChemBioChem, 2009, 10, 799-801
Sieber, S. A.
- "A cyanobacterial serine protease of Plasmodium falciparum is targeted to the apicoplast and plays important role in its growth and development" Mol. Microbiol, 2010
Rathore, S., Sinha, D., Asad, M., Böttcher, T., Afreen, F., Chauhan, V. S., Gupta, D., Sieber, S. A., Mohmmed, A.
(See online at https://doi.org/10.1111/j.1365-2958.2010.07251.x) - "Chemical probes for the labeling of the bacterial glucosaminidase NagZ via the Huisgen cycloaddition" Synthesis, 2010, 13, 2201-2206
Orth, R., Sieber, S.A.
- “Comprehensive Natural Products II: Chemistry and Biology”, Elsevier Science & Technology, 2010, ISBN: 978-0080453811
Sieber, S.A.; Böttcher, T.; Staub, I.; Orth, R.
- “Natural Products and Their Biological Targets: Proteomic and Metabolomic Labeling Strategies” Angewandte Chemie, 2010, 49 (15), 2680-2698
Böttcher, T., Pitscheider, M. and Sieber, S.A.
- “Showdomycin as a versatile chemical tool for the detection of pathogenesis associated enzymes in bacteria” J. Am. Chem. Soc., 2010, 132, 6964-6972
Böttcher, T.; Sieber, S.A.
