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An activity-based probe for antimicrobial target DXP synthase, a thiamin diphosphate-dependent enzyme.
Lauren B Coco1, Caren L Freel Meyers1
1Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Frontiers in Chemical Biology
|November 15, 2024
Summary
Researchers developed a novel alkyl acetylphosphonate (alkylAP) activity-based probe (ABP) to study 1-deoxy-d-xylulose 5-phosphate synthase (DXPS). This probe helps understand DXPS in bacterial infections and aids antimicrobial drug discovery.
Area of Science:
- Biochemistry
- Chemical Biology
- Microbiology
Background:
- 1-deoxy-d-xylulose 5-phosphate synthase (DXPS) is a crucial thiamin diphosphate (ThDP)-dependent enzyme in bacterial metabolism.
- Understanding DXPS's role in pathogen adaptation during infection is limited, with a lack of tools to probe its function in vivo.
Purpose of the Study:
- To develop and characterize a novel activity-based probe (ABP) for DXPS.
- To enable the study of DXPS activity in various bacterial infection contexts and support drug discovery.
Main Methods:
- Design and synthesis of an alkyl acetylphosphonate (alkylAP)-based ABP (compound 1).
- Characterization of ABP 1's reactivity with the ThDP cofactor of DXPS, forming a stable adduct.
- Assessment of probe potency, selectivity, and ability to label DXPS in complex proteomes.
Main Results:
- ABP 1 effectively labels active DXPS with low micromolar potency.
- Labeling is specific to DXPS and can be blocked by alkylAP inhibitors.
- The probe demonstrates selectivity over other ThDP-dependent enzymes and detects DXPS in complex biological samples.
Conclusions:
- The developed alkylAP-based ABP is a valuable tool for investigating DXPS function in bacterial pathogens.
- This probe advances research into DXPS's role in infection and facilitates antimicrobial drug discovery targeting this enzyme.
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