2. Academic Validation
  3. Membrane-active antimicrobial peptide identified in Rana arvalis by targeted DNA sequencing

Membrane-active antimicrobial peptide identified in Rana arvalis by targeted DNA sequencing

  • Biochim Biophys Acta Biomembr. 2019 Mar 1;1861(3):651-659. doi: 10.1016/j.bbamem.2018.12.014.
Tomislav Rončević 1,  Lucija Krce 2,  Marco Gerdol 3,  Sabrina Pacor 4,  Monica Benincasa 5,  Filomena Guida 6,  Ivica Aviani 7,  Vedrana Čikeš-Čulić 8,  Alberto Pallavicini 9,  Ana Maravić 10,  Alessandro Tossi 11
Affiliations

Affiliations

  • 1 Department of Physics, Faculty of Science, University of Split, Croatia. Electronic address: troncevic@pmfst.hr.
  • 2 Department of Physics, Faculty of Science, University of Split, Croatia. Electronic address: lkrce@pmfst.hr.
  • 3 Department of Life Sciences, University of Trieste, Italy. Electronic address: mgerdol@units.it.
  • 4 Department of Life Sciences, University of Trieste, Italy. Electronic address: pacorsab@units.it.
  • 5 Department of Life Sciences, University of Trieste, Italy. Electronic address: mbenincasa@units.it.
  • 6 Department of Life Sciences, University of Trieste, Italy. Electronic address: milenaguida@gmail.com.
  • 7 Department of Physics, Faculty of Science, University of Split, Croatia. Electronic address: iaviani@pmfst.hr.
  • 8 Department of Medical Chemistry and Biochemistry, School of Medicine, University of Split, Croatia. Electronic address: vedrana.cikes.culic@mefst.hr.
  • 9 Department of Life Sciences, University of Trieste, Italy. Electronic address: pallavic@units.it.
  • 10 Department of Biology, Faculty of Science, University of Split, Croatia. Electronic address: amaravic@pmfst.hr.
  • 11 Department of Life Sciences, University of Trieste, Italy. Electronic address: atossi@units.it.
PMID: 30578771 DOI: 10.1016/j.bbamem.2018.12.014
Abstract

Antimicrobial Peptides (AMPs) are naturally produced, gene encoded molecules with a direct antimicrobial activity against pathogens, often also showing Other immune-related properties. Anuran skin secretions are rich in bioactive Peptides, including AMPs, and we have reported a novel targeted Sequencing approach to identify novel AMPs simultaneously in different frog species, from small quantities of skin tissue. Over a hundred full-length Peptides were identified from specimens belonging to five different Ranidae frog species, out of which 29 were novel sequences. Six of these were selected for synthesis and testing against a panel of Gram-negative and Gram-positive bacteria. One peptide, identified in Rana arvalis, proved to be a potent and broad-spectrum antimicrobial, active against ATCC Bacterial strains and a multi-drug resistant clinical isolate. CD spectroscopy suggests it has a helical conformation, while surface plasmon resonance (SPR) that it may self-aggregate/oligomerize at the membrane surface. It was found to disrupt the Bacterial membrane at sub-MIC, MIC and above-MIC concentrations, as observed by flow cytometry and/or visualized by atomic force microscopy (AFM). Only a limited toxicity was observed towards peripheral blood mononuclear cells (PBMC) with a more pronounced effect observed against the MEC-1 cell line.

Keywords

Aggregation; Antibacterial activity; Anuran antimicrobial peptides; Membrane active peptides; Targeted DNA sequencing.

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