Abstract
An in situ screening assay for UDP-galactopyranose mutase (UGM, an essential enzyme of M. tuberculosis cell wall biosynthesis) has been developed to discover novel UGM inhibitors. The approach is based on the amide-forming reaction of an amino acid core with various cinnamic acids, followed by a direct fluorescence polarization assay to identify the best UGM binders without isolation and purification of the screened ligands. This assay allows us to perform one-pot high-throughput synthesis and screening of enzyme inhibitors in a 384-well plate format. UGM ligands were successfully identified by this technology and their inhibition levels were established from pure synthetic compounds in vitro and in a whole cell antibacterial assay. This study provides a blueprint for designing enamide structures as new UGM inhibitors and anti-mycobacterial agents.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acids / chemical synthesis
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Amino Acids / metabolism
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Amino Acids / pharmacology*
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Antitubercular Agents / chemical synthesis
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Antitubercular Agents / metabolism
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Antitubercular Agents / pharmacology*
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Cinnamates / chemical synthesis
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Cinnamates / metabolism
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Cinnamates / pharmacology*
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Enzyme Inhibitors / chemical synthesis
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Enzyme Inhibitors / metabolism
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Enzyme Inhibitors / pharmacology*
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Intramolecular Transferases / antagonists & inhibitors*
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Intramolecular Transferases / chemistry
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Intramolecular Transferases / metabolism
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Kinetics
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Microbial Sensitivity Tests
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Molecular Docking Simulation
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Mycobacterium bovis / drug effects
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Mycobacterium bovis / enzymology
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Mycobacterium tuberculosis / enzymology
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Protein Binding
Substances
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Amino Acids
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Antitubercular Agents
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Cinnamates
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Enzyme Inhibitors
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Intramolecular Transferases
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UDP-galactopyranose mutase