mapping protein–protein interactions of the resistance-related bacterial zeta toxin–epsilon antitoxin complex (ε2ζ2) with high affinity peptide ligands using fluorescence polarization

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ID: 247767
2016
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Abstract
Toxin–antitoxin systems constitute a native survival strategy of pathogenic bacteria and thus are potential targets of antibiotic drugs. Here, we target the Zeta–Epsilon toxin–antitoxin system, which is responsible for the stable maintenance of certain multiresistance plasmids in Gram-positive bacteria. Peptide ligands were designed on the basis of the ε2ζ2 complex. Three α helices of Zeta forming the protein–protein interaction (PPI) site were selected and peptides were designed conserving the residues interacting with Epsilon antitoxin while substituting residues binding intramolecularly to other parts of Zeta. Designed peptides were synthesized with an N-terminal fluoresceinyl-carboxy-residue for binding assays and provided active ligands, which were used to define the hot spots of the ε2ζ2 complex. Further shortening and modification of the binding peptides provided ligands with affinities <100 nM, allowing us to determine the most relevant PPIs and implement a robust competition binding assay.
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fernndez-bachiller2016toxinsmapping Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors ;María Isabel Fernández-Bachiller;Iwona Brzozowska;Norbert Odolczyk;Urszula Zielenkiewicz;Piotr Zielenkiewicz;Jörg Rademann
Journal matec web of conferences
Year 2016
DOI
10.3390/toxins8070222
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