Journal of Agricultural and Food Chemistry · 2019 · 32 citations · 18 references
PharmacyMolecular BiologyPlant PathologyDrug DesignDrug DiscoveryEnzymologyChemical Probe DiscoveryEnzyme InhibitionLead Compound DesignLead DiscoveryBacterial Blight ResistanceBacterial Leaf BlightCell-permeable ProbesClick ChemistryClick Chemistry LabelingNovel Antibacterial Drug DiscoveryStructure-based Lead DiscoveryStructure-based Target Discovery
Target validation of current drugs remains the major challenge for target-based drug discovery, especially for agrochemical discovery. The bactericide 0 represents a novel lead structure and has shown potent efficacy against those diseases that are extremely difficult to control, such as rice bacterial leaf blight. However, no detailed target analysis of this bactericide has been reported. Here, we developed a panel of 0-derived probes 1-6, in which a conservative modification (alkyne tag) was introduced to keep the antibacterial activity of 0 and provide functionality for target identification via click chemistry. With these cell-permeable probes, we were able to discover dihydrolipoamide S-succinyltransferase (DLST) as an unprecedented target in living cells. The probes showed good preference for DLST, especially probe 1, which demonstrated distinct selectivity and reactivity. Also, we reported 0 as the first covalent DLST inhibitor, which has been used to confirm the involvement of DLST in the regulation of energy production.
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In-gel digestion for mass spectrometric characterization of proteins and proteomes
Andrej Shevchenko, Henrik Tomas, J. Havli et al. · Nature Protocols · 2006 · 5.1K citations