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Published on: February 14, 2020
Comparative inhibitory profile and distribution of bacterial PARPs, using Clostridioides difficile CD160 PARP as a
Antonio Ginés García-Saura1,2, Rubén Zapata-Pérez1,2, José Francisco Hidalgo1
1Department of Biochemistry and Molecular Biology-A, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, Campus Espinardo, E-30100, Murcia, Spain.
Abstract:
Poly-ADP-ribose polymerases (PARPs) are involved in the regulation of important cellular processes, such as DNA repair, aging and apoptosis, among others. They have been considered as promising therapeutic targets, since human cancer cells carrying BRCA1 and BRCA2 mutations are highly sensitive to human PARP-1 inhibitors. Although extensive work has been carried out with the latter enzyme, little is known on bacterial PARPs, of which only one has been demonstrated to be active. To extend this limited knowledge, we demonstrate that the Gram-positive bacterium Clostridioides difficile CD160 PARP is a highly active enzyme with a high production yield. Its phylogenetic analysis also pointed to a singular domain organization in contrast to other clostridiales, which could be due to the long-term divergence of C. difficile CD160. Surprisingly, its PARP becomes the first enzyme to be characterized from this strain, which has a genotype never before described based on its sequenced genome. Finally, the inhibition study carried out after a high-throughput in silico screening and an in vitro testing with hPARP1 and bacterial PARPs identified a different inhibitory profile, a new highly inhibitory compound never before described for hPARP1, and a specificity of bacterial PARPs for a compound that mimics NAD+ (EB-47).
Insights
Researchers characterized the highly active Clostridioides difficile CD160 PARP, a novel bacterial enzyme. They discovered a unique inhibitory profile and identified a specific compound targeting bacterial PARPs, distinct from human PARP1 inhibitors.
Area of Science:
- Biochemistry
- Enzymology
- Microbiology
Background:
- Poly-ADP-ribose polymerases (PARPs) regulate crucial cellular processes like DNA repair and apoptosis.
- PARPs are promising therapeutic targets, especially in BRCA-mutated cancers sensitive to PARP inhibitors.
- Knowledge of bacterial PARPs remains limited, with only one previously characterized active enzyme.
Purpose of the Study:
- To characterize the activity and properties of the Clostridioides difficile CD160 PARP.
- To investigate the phylogenetic and domain organization of C. difficile CD160 PARP.
- To explore the inhibitory profiles of compounds against bacterial PARPs and human PARP1.
Main Methods:
- High-throughput in silico screening for potential inhibitors.
- In vitro enzyme inhibition assays using bacterial PARPs and human PARP1.
- Phylogenetic analysis and domain organization studies of C. difficile CD160 PARP.
Main Results:
- C. difficile CD160 PARP was confirmed as a highly active enzyme with a high production yield.
- Phylogenetic analysis revealed a unique domain organization for C. difficile CD160 PARP.
- A novel compound (EB-47) showed high inhibition specificity for bacterial PARPs over human PARP1.
Conclusions:
- C. difficile CD160 PARP is the first characterized enzyme from this unique bacterial strain.
- The distinct domain organization suggests evolutionary divergence in C. difficile CD160.
- The identified compound EB-47 offers potential for developing targeted antibacterial therapies by inhibiting bacterial PARPs.
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