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Visualization of Bacterial Resistance using Fluorescent Antibiotic Probes
Published on: March 2, 2020
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Visualizing the Dynamic Metalation State of New Delhi Metallo-β-lactamase-1 in Bacteria Using a Reversible
Radhika Mehta1, Dann D Rivera2, David J Reilley3
1Department of Chemistry, University of Texas at Austin, 105 East 24th Street Stop A5300, Austin, Texas 78712, United States.
Journal of the American Chemical Society
|May 26, 2021
Summary
A new fluorescent probe tracks the metalation state of New Delhi metallo-β-lactamase (NDM) in E. coli. This tool helps study antibiotic resistance and zinc dyshomeostasis in host-pathogen interactions.
Area of Science:
- Microbiology
- Biochemistry
- Chemical Biology
Background:
- New Delhi metallo-β-lactamase (NDM) confers resistance to critical antibiotics, posing a global health threat.
- Zinc availability influences NDM activity, with variants like NDM-15 showing increased resistance to zinc scarcity.
- Understanding metal ion sequestration in host-pathogen interactions is crucial for combating antibiotic resistance.
Purpose of the Study:
- To develop a novel fluorescent probe for monitoring the dynamic metalation state of NDM within live bacteria.
- To provide a tool for studying host-pathogen interactions and metal ion sequestration.
- To investigate the impact of zinc flux on NDM variants.
Main Methods:
- Development of a thiol-containing, reversible turn-on fluorescent probe.
- Selective coordination of the dizinc metal cluster in NDM, leading to a significant fluorescence increase.
- Confocal microscopy imaging of live *Escherichia coli* to visualize NDM localization and metalation state.
Main Results:
- The probe demonstrated a 17-fold increase in fluorescence upon binding to NDM.
- The probe enabled detection of enzyme localization, substrate/inhibitor engagement, and metalation changes in live *E. coli*.
- NDM-1 was susceptible to demetalation by chelators, while NDM-15 showed greater resistance to zinc flux.
Conclusions:
- The developed fluorescent probe is a valuable tool for studying NDM metalation dynamics in real-time.
- This probe facilitates research into host defense mechanisms affecting metal ion availability and NDM activity.
- It aids in the development of therapeutics by enabling detection of inhibitor-target engagement.
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