title: Human β-defensin 4 with non-native disulfide bridges exhibit antimicrobial activity.
abstract: Human defensins play multiple roles in innate immunity including direct antimicrobial killing and immunomodulatory activity. They have three disulfide bridges which contribute to the stability of three anti-parallel β-strands. The exact role of disulfide bridges and canonical β-structure in the antimicrobial action is not yet fully understood. In this study, we have explored the antimicrobial activity of human β-defensin 4 (HBD4) analogs that differ in the number and connectivity of disulfide bridges. The cysteine framework was similar to the disulfide bridges present in μ-conotoxins, an unrelated class of peptide toxins. All the analogs possessed enhanced antimicrobial potency as compared to native HBD4. Among the analogs, the single disulfide bridged peptide showed maximum potency. However, there were no marked differences in the secondary structure of the analogs. Subtle variations were observed in the localization and membrane interaction of the analogs with bacteria and Candida albicans, suggesting a role for disulfide bridges in modulating their antimicrobial action. All analogs accumulated in the cytosol where they can bind to anionic molecules such as nucleic acids which would affect several cellular processes leading to cell death. Our study strongly suggests that native disulfide bridges or the canonical β-strands in defensins have not evolved for maximal activity but they play important roles in determining their antimicrobial potency.
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology, Hyderabad, Telangana, India.
authors.forename: Himanshu
authors.initials: H
authors.lastname: Sharma
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology, Hyderabad, Telangana, India.
authors.forename: Ramakrishnan
authors.initials: R
authors.lastname: Nagaraj
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure; D020916:Conotoxins / Q000737:chemistry; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry*; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure; D006801:Humans; D007113:Immunity, Innate; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D017433:Protein Structure, Secondary; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure; D060327:Solid-Phase Synthesis Techniques; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure; D023083:beta-Defensins / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D020916:Conotoxins; C106641:DEFB4A protein, human; D004220:Disulfides; D010455:Peptides; D023083:beta-Defensins