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Characterization of Lipidated New Delhi Metallo-β-lactamase Using Synthetic Nanodiscs
Thomas Smisek1, Nemanja Vuksanovic2, Jada N Walker3
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712, United States.
Biochemistry
|June 3, 2025
Summary
New Delhi metallo-β-lactamase (NDM) is unique due to membrane anchoring. This study optimized its expression and used nanodiscs to show membrane anchoring enhances zinc affinity, aiding bacterial survival.
Area of Science:
- Biochemistry
- Microbiology
- Structural Biology
Background:
- Carbapenem-resistant bacteria pose a significant global health threat.
- Metallo-carbapenemases, like New Delhi metallo-β-lactamase (NDM), are enzymes conferring resistance to most β-lactam antibiotics.
- NDM is unique among metallo-carbapenemases for its membrane-anchored structure, unlike soluble periplasmic forms.
Purpose of the Study:
- To investigate the functional significance of NDM's unique membrane anchoring.
- To characterize the biochemical properties of lipidated NDM in a membrane-like environment.
- To establish a novel platform for studying bacterial lipoproteins.
Main Methods:
- Optimized overexpression of lipidated NDM in *Escherichia coli*.
- Solubilized the NDM lipoprotein into synthetic nanodiscs for biochemical analysis.
- Assessed zinc affinity and thermostability of NDM variants.
Main Results:
- Membrane anchoring was found to enhance NDM's zinc affinity, suggesting adaptation to zinc-limiting environments.
- Thermostability varied across different clinical NDM variants.
- Successfully established a nanodisc-based system for studying membrane-bound NDM.
Conclusions:
- Membrane anchoring is a key adaptation for NDM, improving its fitness under specific environmental conditions.
- The nanodisc platform provides a viable method for studying the function of membrane-anchored bacterial lipoproteins.
- Further research into NDM variants and their interactions is warranted.
