The "superbug" infection caused by New Delhi metallo-β-lactamase (NDM-1) has become an emerging threat. Monitoring NDM-1 has proven challenging due to its shuttling between pathogenic bacteria. Here, we report an isothermal titration calorimetry (ITC) method that can monitor activity and inhibition of NDM-1 in live bacterial cells in real time. This method has been exemplified by monitoring of the activity and inhibition of the target enzyme and evaluating the breakdown of antibiotics by pathogenic bacteria expressing β-lactamases. Cell-based studies demonstrate that the NDM-1 expressed in bacterial cells was inhibited by four known inhibitors ethylene diamine tetraacetic acid (EDTA), d-captopril, ebselen and azolylthioacetamide with fifty percent inhibitory concentration (IC) values of 3.8, 48, 0.55, and 17.5 μM, respectively, which are in good agreement with the data from inhibition kinetics using UV-vis and NMR spectroscopy in vivo. This approach could be applied to screen and evaluate small molecule inhibitors of metallo-β-lactamases (MβLs) in whole cells or to identify drug resistant bacteria.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1021/acsinfecdis.8b00147 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Potent antibacterial and cytotoxic bioactive compounds from endophytic fungi Diaporthe sp. associated with Salacia intermedia.
Arch Microbiol
January 2025
Research Center for Pharmaceutical Ingredients and Traditional Medicine, National Research and Innovation Agency (BRIN), KST B.J. Habibie, Serpong, South Tangerang, 15314, Indonesia.
Antibacterial screening of endophytic fungi from Salacia intermedia identified Diaporthe longicolla as a potent strain exhibiting good activity against multidrug-resistant Staphylococcus aureus and Pseudomonas aeruginosa, with an MIC of 39.1 µg/mL. Scale-up fermentation and chromatographic purification of this strain yielded three known compounds, which were cytochalasin J (1), cytochalasin H (2), and dicerandrol C (3), as identified by liquid chromatography - high mass resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy.
View Article and Find Full Text PDFItaconate mechanism of action and dissimilation in .
Proc Natl Acad Sci U S A
January 2025
Centre for Tuberculosis Research, Tuberculosis Research Laboratory, Translational Health Science and Technology Institute, National Capital Region Biotech Science Cluster 3rd Milestone, Faridabad, Haryana 121001, India.
Itaconate, an abundant metabolite produced by macrophages upon interferon-γ stimulation, possesses both antibacterial and immunomodulatory properties. Despite its crucial role in immunity and antimicrobial control, its mechanism of action and dissimilation are poorly understood. Here, we demonstrate that infection of mice with increases itaconate levels in lung tissues.
View Article and Find Full Text PDFA periplasmic protein modulates the proteolysis of peptidoglycan hydrolases to maintain cell wall homeostasis in .
Proc Natl Acad Sci U S A
January 2025
Department of Biological Sciences, College of Natural Sciences, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Bacterial cell wall assembly and remodeling require activities of peptidoglycan (PG) hydrolases as well as PG synthases. In particular, the activity of DD-endopeptidases, which cleave the 4-3 peptide crosslinks in PG, is essential for PG expansion in gram-negative bacteria. Maintaining optimal levels of DD-endopeptidases is critical for expanding PG without compromising its integrity.
View Article and Find Full Text PDFIdentification of 6,8-ditrifluoromethyl halogenated phenazine as a potent bacterial biofilm-eradicating agent.
Org Biomol Chem
January 2025
Department of Pharmaceutical & Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, USA.
Bacterial biofilms are surface-attached communities consisting of non-replicating persister cells encased within an extracellular matrix of biomolecules. Unlike bacteria that have acquired resistance to antibiotics, persister cells enable biofilms to demonstrate innate tolerance toward all classes of conventional antibiotic therapies. It is estimated that 50-80% of bacterial infections are biofilm associated, which is considered the underlying cause of chronic and recurring infections.
View Article and Find Full Text PDFUncovering a new family of conserved virulence factors that promote the production of host-damaging outer membrane vesicles in gram-negative bacteria.
J Extracell Vesicles
January 2025
Institut de Recherche en Santé Digestive (IRSD), Université de Toulouse, INSERM, INRAE, ENVT, UPS, Toulouse, France.
CprA is a short-chain dehydrogenase/reductase (SDR) that contributes to resistance against colistin and antimicrobial peptides. The cprA gene is conserved across Pseudomonas aeruginosa clades and its expression is directly regulated by the two-component system PmrAB. We have shown that cprA expression leads to the production of outer membrane vesicles (OMVs) that block autophagic flux and have a greater capacity to activate the non-canonical inflammasome pathway.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!