Structural and Functional Studies of Bacterial Enolase, a Potential Target against Gram-Negative Pathogens

Enolase is a glycolytic metalloenzyme involved in carbon metabolism. The advantage of targeting enolase lies in its essentiality in many biological processes such as cell wall formation and RNA turnover and as a plasminogen receptor. We initially used a DARTS assay to identify enolase as a target in...

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Bibliographic Details
Published in:Biochemistry (Easton) 2019-03, Vol.58 (9), p.1188-1197
Main Authors: Krucinska, Jolanta, Falcone, Eric, Erlandsen, Heidi, Hazeen, Akram, Lombardo, Michael N, Estrada, Alexavier, Robinson, Victoria L, Anderson, Amy C, Wright, Dennis L
Format: Article
Language:English
Subjects:
Anti-Bacterial Agents - chemistry
Anti-Bacterial Agents - pharmacology
Calorimetry
Catalytic Domain
Crystallography, X-Ray
Drug Evaluation, Preclinical
Enzyme Inhibitors - chemistry
Enzyme Inhibitors - pharmacology
Escherichia coli Proteins - antagonists & inhibitors
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - genetics
Escherichia coli Proteins - metabolism
Gram-Negative Bacteria - drug effects
Gram-Negative Bacteria - enzymology
Microbial Sensitivity Tests
Molecular Docking Simulation
Phosphopyruvate Hydratase - antagonists & inhibitors
Phosphopyruvate Hydratase - chemistry
Phosphopyruvate Hydratase - genetics
Phosphopyruvate Hydratase - metabolism
Protein Conformation
Structure-Activity Relationship
Tropolone - chemistry
Tropolone - pharmacology
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Summary:Enolase is a glycolytic metalloenzyme involved in carbon metabolism. The advantage of targeting enolase lies in its essentiality in many biological processes such as cell wall formation and RNA turnover and as a plasminogen receptor. We initially used a DARTS assay to identify enolase as a target in Escherichia coli. The antibacterial activities of α-, β-, and γ-substituted seven-member ring tropolones were first evaluated against four strains representing a range of Gram-negative bacteria. We observed that the chemical properties and position of the substituents on the tropolone ring play an important role in the biological activity of the investigated compounds. Both α- and β-substituted phenyl derivatives of tropolone were the most active with minimum inhibitory concentrations in the range of 11–14 μg/mL. The potential inhibitory activity of the synthetic tropolones was further evaluated using an enolase inhibition assay, X-ray crystallography, and molecular docking simulations. The catalytic activity of enolase was effectively inhibited by both the naturally occurring β-thujaplicin and the α- and β-substituted phenyl derivatives of tropolones with IC50 values in range of 8–11 μM. Ligand binding parameters were assessed by isothermal titration calorimetry and differential scanning calorimetry techniques and agreed with the in vitro data. Our studies validate the antibacterial potential of tropolones with careful consideration of the position and character of chelating moieties for stronger interaction with metal ions and residues in the enolase active site.
ISSN:0006-2960
1520-4995
DOI:10.1021/acs.biochem.8b01298