title: D-Amino acids incorporation in the frog skin-derived peptide esculentin-1a(1-21)NH2 is beneficial for its multiple functions.
abstract: Naturally occurring antimicrobial peptides (AMPs) represent promising future antibiotics. We have previously isolated esculentin-1a(1-21)NH2, a short peptide derived from the frog skin AMP esculentin-1a, with a potent anti-Pseudomonal activity. Here, we investigated additional functions of the peptide and properties responsible for these activities. For that purpose, we synthesized the peptide, as well as its structurally altered analog containing two D-amino acids. The peptides were then biophysically and biologically investigated for their cytotoxicity and immunomodulating activities. The data revealed that compared to the wild-type, the diastereomer: (1) is significantly less toxic towards mammalian cells, in agreement with its lower α-helical structure, as determined by circular dichroism spectroscopy; (2) is more effective against the biofilm form of Pseudomonas aeruginosa (responsible for lung infections in cystic fibrosis sufferers), while maintaining a high activity against the free-living form of this important pathogen; (3) is more stable in serum; (4) has a higher activity in promoting migration of lung epithelial cells, and presumably in healing damaged lung tissue, and (5) disaggregates and detoxifies the bacterial lipopolysaccharide (LPS), albeit less than the wild-type. Light scattering studies revealed a correlation between anti-LPS activity and the ability to disaggregate the LPS. Besides shedding light on the multifunction properties of esculentin-1a(1-21)NH2, the D-amino acid containing isomer may serve as an attractive template for the development of new anti-Pseudomonal compounds with additional beneficial properties. Furthermore, together with other studies, incorporation of D-amino acids may serve as a general approach to optimize the future design of new AMPs.
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Di Grazia
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy.
authors.forename: Floriana
authors.initials: F
authors.lastname: Cappiello
authors.affiliation: Department of Biological Chemistry, The Weizmann Institute of Science, 76100, Rehovot, Israel.
authors.forename: Hadar
authors.initials: H
authors.lastname: Cohen
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy.
authors.forename: Bruno
authors.initials: B
authors.lastname: Casciaro
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Luca
authors.affiliation: Department of Medical Biotechnology, University of Siena, 53100, Siena, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pini
authors.affiliation: Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, 15260, USA.
authors.forename: Y Peter
authors.initials: YP
authors.lastname: Di
authors.affiliation: Department of Biological Chemistry, The Weizmann Institute of Science, 76100, Rehovot, Israel.
authors.forename: Yechiel
authors.initials: Y
authors.lastname: Shai
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy. marialuisa.mangoni@uniroma1.it.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Mangoni
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D029845:Amphibian Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D018441:Biofilms / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival; D002942:Circular Dichroism; D004847:Epithelial Cells / Q000187:drug effects; D006801:Humans; D008070:Lipopolysaccharides / Q000737:chemistry; D008264:Macrophages / Q000187:drug effects; D051379:Mice; D008826:Microbial Sensitivity Tests; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000187:drug effects*; D011650:Pulmonary Alveoli / Q000166:cytology; D000067996:RAW 264.7 Cells; D011898:Ranidae; D012867:Skin / Q000737:chemistry; D013237:Stereoisomerism
chemical_list: D000596:Amino Acids; D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides; C092874:esculentin protein, Rana esculenta
keywords: Anti-biofilm activity; Antimicrobial peptide; Diastereomer; Esculentin-1; Wound healing