Identification of a novel antimicrobial peptide, scolopendin 1, derived from centipede Scolopendra subspinipes mutilans and its antifungal mechanism

Insect Mol Biol. 2014 Dec;23(6):788-99. doi: 10.1111/imb.12124. Epub 2014 Sep 11.

Abstract

In this study, a novel antimicrobial peptide, scolopendin 1, was identified from adult centipedes, Scolopendra subspinipes mutilans using RNA sequencing. Scolopendin 1 exerted an antimicrobial activity without inducing haemolysis of human erythrocytes. In order to understand the antifungal mechanism, a reactive oxygen species (ROS) assay was performed, which indicated that scolopendin 1 induced ROS accumulation in Candida albicans. Evaluation of fungal viability using N-acetyl cysteine, a ROS scavenger, suggested that ROS are a major factor in scolopendin 1-induced fungal cell death. Co-staining of annexin V-fluorescein isothiocyanate (FITC) and propidium iodide, and TUNEL and 4',6-diamidino-2-phenylindole (DAPI) assays confirmed that ROS-induced fungal cell death is associated with apoptosis. To further investigate the mechanism that facilitates the progression of apoptosis, changes in intracellular Ca(2+) concentration and mitochondrial dysfunction were examined. Ca(2+) , a signalling molecule in the apoptotic pathway, was increased in the cytosol and mitochondria, and ROS accumulation triggered mitochondrial depolarization and the release of cytochrome c, a pro-apoptotic factor, from the mitochondria to the cytosol. Finally, the released cytochrome c activated intracellular caspase. The present study suggests that scolopendin 1 could emerge as a model molecule that targets the apoptotic pathway and provides a novel remedy.

Keywords: RNA sequencing; Scolopendra subspinipes mutilans; antimicrobial peptide; scolopendin 1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antifungal Agents / chemical synthesis
  • Antifungal Agents / chemistry
  • Antifungal Agents / pharmacology*
  • Antimicrobial Cationic Peptides / chemical synthesis
  • Antimicrobial Cationic Peptides / chemistry
  • Antimicrobial Cationic Peptides / genetics*
  • Antimicrobial Cationic Peptides / isolation & purification
  • Antimicrobial Cationic Peptides / pharmacology*
  • Apoptosis / drug effects
  • Arthropod Proteins / genetics
  • Arthropod Proteins / pharmacology*
  • Arthropods / genetics*
  • Arthropods / immunology
  • Arthropods / microbiology
  • Candida albicans / drug effects
  • Candida albicans / growth & development
  • Caspases
  • Erythrocytes / drug effects
  • Escherichia coli
  • Hemolysis / drug effects
  • Humans
  • Microbial Sensitivity Tests
  • Mitochondria / drug effects
  • Molecular Sequence Data
  • Reactive Oxygen Species / metabolism

Substances

  • Antifungal Agents
  • Antimicrobial Cationic Peptides
  • Arthropod Proteins
  • Reactive Oxygen Species
  • scolopendin 1, Scolopendra subspinipes
  • Caspases