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The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
Published on: February 1, 2018
Structural and kinetic studies on metallo-β-lactamase IMP-1
Dionne H Griffin1, Timothy K Richmond, Carlo Sanchez
1Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056, United States.
Biochemistry
|September 21, 2011
Summary
Metallo-β-lactamase (MβL) IMP-1 binds zinc and exhibits catalytic activity. Metal binding to IMP-1 follows a positive-cooperative mechanism, as indicated by kinetic and spectroscopic studies.
Area of Science:
- Biochemistry
- Enzymology
- Metalloprotein research
Background:
- Metallo-β-lactamases (MβLs) are crucial in antibiotic resistance.
- Understanding metal ion coordination in MβL IMP-1 is key to its function.
Purpose of the Study:
- To investigate metal binding mechanisms in metallo-β-lactamase (MβL) IMP-1.
- To characterize the catalytic activity and metal coordination of IMP-1.
Main Methods:
- Enzyme overexpression, purification, and characterization.
- Extended X-ray absorption fine structure (EXAFS) and UV-visible spectroscopy.
- Electron paramagnetic resonance (EPR) and rapid kinetic studies.
Main Results:
- IMP-1 binds 2 equivalents of Zn(II) and displays significant catalytic activity.
- Kinetic studies reveal a simple Michaelis mechanism for nitrocefin hydrolysis.
- Spectroscopic data suggest a positive-cooperative metal binding mode to metal-free IMP-1.
Conclusions:
- Metal binding to IMP-1 is a positive-cooperative process.
- The findings provide insights into the metalloenzyme's active site and mechanism.
- This study contributes to understanding MβL function and potential inhibition strategies.
