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Identification of Inhibitors Targeting Ferredoxin-NADP⁺ Reductase from the Xanthomonas citri subsp. citri Phytopathogenic Bacteria.
Ferredoxin-NADP(H) reductases (FNRs) deliver NADPH or low potential one-electron donors to redox-based metabolism in plastids and bacteria. Xanthomonas citri subsp. citri ( Xcc ) is a Gram-negative bacterium responsible for citrus canker disease that affects commercial citrus crops worldwide. The Xcc fpr gene encodes a bacterial type FNR ( Xcc FPR) that contributes to the bacterial response to oxidative stress conditions, usually found during plant colonization. Therefore, Xcc FPR is relevant for the pathogen survival and its inhibition might represent a strategy to treat citrus canker. Because of mechanistic and structural differences from plastidic FNRs, Xcc FPR is also a potential antibacterial target. We have optimized an activity-based high-throughput screening (HTS) assay that identifies Xcc FPR inhibitors. We selected 43 hits from a chemical library and narrowed them down to the four most promising inhibitors. The antimicrobial effect of these compounds was evaluated on Xcc cultures, finding one with antimicrobial properties. Based on the functional groups of this compound and their geometric arrangement, we identified another three Xcc FPR inhibitors. Inhibition mechanisms and constants were determined for these four Xcc FPR inhibitors. Their specificity was also evaluated by studying their effect on the plastidic Anabaena PCC 7119 FNR, finding differences that can become interesting tools to discover Xcc antimicrobials.
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