Nuclease activity gives an edge to host‐defense peptide piscidin 3 over piscidin 1, rendering it more effective against persisters and biofilms

• Published in: The FEBS journal 2017-11, Vol.284 (21), p.3662-3683
• Main Authors: Libardo, M. Daben J., Bahar, Ali A., Ma, Buyong, Fu, Riqiang, McCormick, Laura E., Zhao, Jun, McCallum, Scott A., Nussinov, Ruth, Ren, Dacheng, Angeles‐Boza, Alfredo M., Cotten, Myriam L.
• Format: Article
• Language: English
• Subjects: Animal tissues; Animals; Anti-Bacterial Agents - isolation & purification; Anti-Bacterial Agents - metabolism; Anti-Bacterial Agents - pharmacology; Antimicrobial Cationic Peptides - isolation & purification; Antimicrobial Cationic Peptides - metabolism; antimicrobial peptide; Bacteria; Binding; Biofilms; Biofilms - drug effects; Copper; Deoxyribonucleic acid; DNA; DNA Cleavage - drug effects; DNA damage; Eradication; Escherichia coli - cytology; Escherichia coli - drug effects; Escherichia coli - growth & development; Fish Proteins - isolation & purification; Fish Proteins - metabolism; Fishes; Homology; host defense; Hydrophobicity; Microbial Sensitivity Tests; Molecular dynamics; Molecular Dynamics Simulation; Nickel; NMR; Nuclear magnetic resonance; Nuclease; Peptides; Physicochemical properties; Pseudomonas aeruginosa - cytology; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - growth & development; reactive oxygen species; Staphylococcus aureus - cytology; Staphylococcus aureus - drug effects; Staphylococcus aureus - growth & development
• ISSN: 1742-464X; 1742-4658; 1742-4658
• DOI: 10.1111/febs.14263

Host‐defense peptides (HDPs) feature evolution‐tested potency against life‐threatening pathogens. While piscidin 1 (p1) and piscidin 3 (p3) are homologous and potent fish HDPs, only p1 is strongly membranolytic. Here, we hypothesize that another mechanism imparts p3 strong potency. We demonstrate that the N‐termini of both peptides coordinate Cu2+ and p3‐Cu cleaves isolated DNA at a rate on par with free Cu2+ but significantly faster than p1‐Cu. On planktonic bacteria, p1 is more antimicrobial but only p3 features copper‐dependent DNA cleavage. On biofilms and persister cells, p3‐Cu is more active than p1‐Cu, commensurate with stronger peptide‐induced DNA damage. Molecular dynamics and NMR show that more DNA‐peptide interactions exist with p3 than p1, and the peptides adopt conformations simultaneously poised for metal‐ and DNA‐binding. These results generate several important conclusions. First, homologous HDPs cannot be assumed to have identical mechanisms since p1 and p3 eradicate bacteria through distinct relative contributions of membrane and DNA‐disruptive effects. Second, the nuclease and membrane activities of p1 and p3 show that naturally occurring HDPs can inflict not only physicochemical but also covalent damage. Third, strong nuclease activity is essential for biofilm and persister cell eradication, as shown by p3, the homolog more specific toward bacteria and more expressed in vascularized tissues. Fourth, p3 combines several physicochemical properties (e.g., Amino Terminal Copper and Nickel binding motif; numerous arginines; moderate hydrophobicity) that confer low membranolytic effects, robust copper‐scavenging capability, strong interactions with DNA, and fast nuclease activity. This new knowledge could help design novel therapeutics active against hard‐to‐treat persister cells and biofilms. Piscidins are innate immune factors from several fish species. We show that piscidins 1 and 3 bind to copper ions through their N‐terminal domain to combat bacteria via oxidative damage. Piscidin 3 is more efficient at DNA oxidation resulting in more effective biofilm and persister cell eradication. These mechanistic, which studies that document peptide‐copper synergy, advance our understanding of vertebrate immunity.