Dataset LC MS Manool disarms Streptococcus mutans by targeting ABC trasporters protein
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| Natura: | Recurso digital |
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Zenodo
2025
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| _version_ | 1866901280860930048 |
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| author | Nocera, Raffaella Donadio, Giuliana |
| author_facet | Nocera, Raffaella Donadio, Giuliana |
| contents | <p>The emergency of antibiotic resistance is serious. We must identify molecules that escape the self-defense mechanisms of microorganisms. In this study, we optimized a protocol for identifying Streptococcus mutans proteins impaired by manool, a metabolite from Salvia officinalis with antimicrobial activity. The research showed that S. mutans can evade antibiotics and grow. This study shows the potential of manool as a new antimicrobial agent and the importance of understanding the molecular mechanisms of plant-derived compounds in the fight against AMR. Samples were obtained using the Drug Affinity Responsive Target Stability (DART-S) assay. A new protocol was performed for bacterial cells that allows the DARTS assay to be applied to a prokaryotic model using a minimal medium. The DARTS assay was initially performed by incubating manool with a protein extract (pe-DARTS) from S. mutans, followed by limited proteolysis, SDS-PAGE analysis of digestion products, and proteomic identification of the separated proteins. The bc-DARTS assay involved the incubation of manool with Streptococcus mutans bacterial cells, followed by proteolysis, SDS-PAGE analysis, and LC-MS. The S. mutans overnight inoculum was centrifuged at 3000 rpm for 15 minutes, and the cells were resuspended in BHI medium to a density of 0.05 OD/ml. The cells were incubated at 37°C and cultured to the established optical densities (0.5 OD/ml and 1 OD/ml). Then cells were centrifuged at 3000 rpm for 15 minutes. After discarding the supernatant, the pellet was resuspended in M9 minimal medium 1X supplemented with 0.4% glucose for both control and treated samples, at a density of 0,5 OD/ml and 1 OD/mL. The treated samples were incubated with manool at 5 µM and 10 µM for 90 minutes.<br><br></p> |
| format | Recurso digital |
| id | zenodo_https___doi_org_10_5281_zenodo_15743861 |
| institution | Zenodo |
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| publishDate | 2025 |
| publisher | Zenodo |
| record_format | zenodo |
| spellingShingle | Dataset LC MS Manool disarms Streptococcus mutans by targeting ABC trasporters protein Nocera, Raffaella Donadio, Giuliana <p>The emergency of antibiotic resistance is serious. We must identify molecules that escape the self-defense mechanisms of microorganisms. In this study, we optimized a protocol for identifying Streptococcus mutans proteins impaired by manool, a metabolite from Salvia officinalis with antimicrobial activity. The research showed that S. mutans can evade antibiotics and grow. This study shows the potential of manool as a new antimicrobial agent and the importance of understanding the molecular mechanisms of plant-derived compounds in the fight against AMR. Samples were obtained using the Drug Affinity Responsive Target Stability (DART-S) assay. A new protocol was performed for bacterial cells that allows the DARTS assay to be applied to a prokaryotic model using a minimal medium. The DARTS assay was initially performed by incubating manool with a protein extract (pe-DARTS) from S. mutans, followed by limited proteolysis, SDS-PAGE analysis of digestion products, and proteomic identification of the separated proteins. The bc-DARTS assay involved the incubation of manool with Streptococcus mutans bacterial cells, followed by proteolysis, SDS-PAGE analysis, and LC-MS. The S. mutans overnight inoculum was centrifuged at 3000 rpm for 15 minutes, and the cells were resuspended in BHI medium to a density of 0.05 OD/ml. The cells were incubated at 37°C and cultured to the established optical densities (0.5 OD/ml and 1 OD/ml). Then cells were centrifuged at 3000 rpm for 15 minutes. After discarding the supernatant, the pellet was resuspended in M9 minimal medium 1X supplemented with 0.4% glucose for both control and treated samples, at a density of 0,5 OD/ml and 1 OD/mL. The treated samples were incubated with manool at 5 µM and 10 µM for 90 minutes.<br><br></p> |
| title | Dataset LC MS Manool disarms Streptococcus mutans by targeting ABC trasporters protein |
| url | https://doi.org/10.5281/zenodo.15743861 |