FN Archimer Export Format PT J TI The Ancestral N-Terminal Domain of Big Defensins Drives Bacterially Triggered Assembly into Antimicrobial Nanonets BT AF Loth, Karine Vergnes, Agnes Barreto, Cairé Voisin, Sébastien N Meudal, Hervé Da Silva, Jennifer Bressan, Albert Belmadi, Nawal Bachère, Evelyne Aucagne, Vincent Cazevielle, Chantal Marchandin, Hélène Rosa, Rafael Diego Bulet, Philippe Touqui, Lhousseine Delmas, Agnès F. Destoumieux-Garzón, Delphine AS 1:1,2;2:3;3:3,4;4:5;5:1;6:6;7:3,4;8:6;9:3;10:1;11:7;12:8;13:4;14:5,9;15:6;16:1;17:10; FF 1:;2:PDG-RBE-IHPE;3:;4:;5:;6:;7:;8:;9:PDG-RBE-IHPE;10:;11:;12:;13:;14:;15:;16:;17:; C1 Centre de Biophysique Moléculaire UPR4301 CNRS, Orléans, France UFR CoST, Université d’Orléans, Orléans, France IHPE, Université de Montpellier, CNRS, Ifremer, Université de Perpignan Via Domitia, Montpellier, France Laboratory of Immunology Applied to Aquaculture, Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil Plateforme BioPark d’Archamps, Archamps Technopole, Archamps, France Equipe mixte Institut Pasteur/Paris V Mucoviscidose et Bronchopathies Chroniques, Institut Pasteur, Paris, France COMET, Plateau de microscopie électronique, Plateforme Montpellier RIO Imaging, Montpellier, France HydroSciences Montpellier, Département de Microbiologie, CHU Nîmes, CNRS, IRD, Université de Montpellier, Nîmes, France Institute for Advanced Biosciences, CR Université Grenoble Alpes, CNRS UMR5309, La Tronche, France IHPE, Université de Montpellier, CNRS, Ifremer, Université de Perpignan Via Domitia, Montpellier, France C2 CNRS, FRANCE UNIV ORLEANS, FRANCE IFREMER, FRANCE UNIV FED SANTA CATARINA, BRAZIL BIOPARK, FRANCE INST PASTEUR, FRANCE INSERM, FRANCE UNIV MONTPELLIER, FRANCE CNRS, FRANCE CNRS, FRANCE SI MONTPELLIER SE PDG-RBE-IHPE UM IHPE IN WOS Ifremer UMR WOS Cotutelle UMR DOAJ copubli-france copubli-univ-france copubli-int-hors-europe copubli-sud IF 6.784 TC 29 UR https://archimer.ifremer.fr/doc/00588/70057/68000.pdf LA English DT Article DE ;MRSA;antimicrobial peptides;antimicrobial resistance;defensins;fibrils;innate immunity;mechanisms of action;nuclear magnetic resonance AB Big defensins, ancestors of β-defensins, are composed of a β-defensin-like C-terminal domain and a globular hydrophobic ancestral N-terminal domain. This unique structure is found in a limited number of phylogenetically distant species, including mollusks, ancestral chelicerates, and early-branching cephalochordates, mostly living in marine environments. One puzzling evolutionary issue concerns the advantage for these species of having maintained a hydrophobic domain lost during evolution toward β-defensins. Using native ligation chemistry, we produced the oyster Crassostrea gigas BigDef1 (Cg-BigDef1) and its separate domains. Cg-BigDef1 showed salt-stable and broad-range bactericidal activity, including against multidrug-resistant human clinical isolates of Staphylococcus aureus. We found that the ancestral N-terminal domain confers salt-stable antimicrobial activity to the β-defensin-like domain, which is otherwise inactive. Moreover, upon contact with bacteria, the N-terminal domain drives Cg-BigDef1 assembly into nanonets that entrap and kill bacteria. We speculate that the hydrophobic N-terminal domain of big defensins has been retained in marine phyla to confer salt-stable interactions with bacterial membranes in environments where electrostatic interactions are impaired. Those remarkable properties open the way to future drug developments when physiological salt concentrations inhibit the antimicrobial activity of vertebrate β-defensins. IMPORTANCE β-Defensins are host defense peptides controlling infections in species ranging from humans to invertebrates. However, the antimicrobial activity of most human β-defensins is impaired at physiological salt concentrations. We explored the properties of big defensins, the β-defensin ancestors, which have been conserved in a number of marine organisms, mainly mollusks. By focusing on a big defensin from oyster (Cg-BigDef1), we showed that the N-terminal domain lost during evolution toward β-defensins confers bactericidal activity to Cg-BigDef1, even at high salt concentrations. Cg-BigDef1 killed multidrug-resistant human clinical isolates of Staphylococcus aureus. Moreover, the ancestral N-terminal domain drove the assembly of the big defensin into nanonets in which bacteria are entrapped and killed. This discovery may explain why the ancestral N-terminal domain has been maintained in diverse marine phyla and creates a new path of discovery to design β-defensin derivatives active at physiological and high salt concentrations. PY 2019 PD SEP SO Mbio SN 2150-7511 PU American Society for Microbiology VL 10 IS 5 UT 000493915800090 DI 10.1128/mBio.01821-19 ID 70057 ER EF