Characteristics of Novel Insect Defensin-Based Membrane-Disrupting Trypanocidal Peptides

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  • YAMAGE Mat
    Innate Immunity Research Unit, National Institute of the Agrobiological Sciences
  • YOSHIYAMA Mikio
    Innate Immunity Research Unit, National Institute of the Agrobiological Sciences
  • GRAB Dennis J.
    Department of Pediatrics, The Johns Hopkins School of Medicine
  • KUBO Masanori
    Department of Epidemiology, National Institute of Animal Health
  • IWASAKI Takashi
    Graduate School of Life and Environmental Sciences, University of Tsukuba
  • KITANI Hiroshi
    Transgenic Animal Research Center, National Institute of the Agrobiological Sciences
  • ISHIBASHI Jun
    Innate Immunity Research Unit, National Institute of the Agrobiological Sciences
  • YAMAKAWA Minoru
    Innate Immunity Research Unit, National Institute of the Agrobiological Sciences Graduate School of Life and Environmental Sciences, University of Tsukuba

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Synthetic D- and L-amino acid type cationic 9-mer peptides (all sequences were synthesized as D- or L-amino acids) derived from the active sites of insect defensins were tested for their ability to modify the growth of blood-stream form African trypanosomes in vitro. One of them, the D-type peptide A (RLYLRIGRR-NH2), irreversibly suppressed proliferation of the Trypanosoma brucei brucei GUTat3.1 parasite. The presence of negatively charged phosphatidylserine on the surface of the parasites was demonstrated, suggesting electrostatic interaction between the peptide and the phospholipids. Furthermore, this peptide was found to alter trypanosome membrane-potentials significantly, an effect apparently due to the removal of the parasite’s plasma membrane. The potential toxic effects of D-peptide A on mammalian cells was assessed using human brain microvascular endothelial cells. Only minor effects were found when the endothelial cells were exposed for 16 h to peptide concentrations of less than 200 μM. These findings suggest that insect defensin-based peptides represent a potentially new class of membrane-disrupting trypanocidal drugs.

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