The Synergistic Antimicrobial Effects of Novel Bombinin and Bombinin H Peptides from the Skin Secretion of Bombina orientalis

Jie Xiang, Mei Zhou, Yuxin Wu, Tianbao Chen, Christopher Shaw, Lei Wang

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Abstract

Bombinin and bombinin H are two antimicrobial peptide (AMP) families initially discovered from the skin secretion of Bombina that share the same biosynthetic precursor-encoding cDNAs, but have different structures and physicochemical properties. Insight into their possible existing relationship lead us to perform the combination investigations into their anti-infectious activities. In this work, we report the molecular cloning and functional characterisation of two novel AMPs belonging to bombinin and bombinin H families from secretions of Bombina orientalis . Their mature peptides (BHL-bombinin and bombinin HL), coded by single open-reading frame (ORF), were chemically synthesised along with an analogue peptide that replaced L-leucine with D-leucine from the 2nd position of the N-terminus (bombinin HD). Circular dichroism analysis revealed that all of them displayed well-defined α-helical structures in membrane mimicking environments. Furthermore, BHL-bombinin displayed broad-spectrum bactericidal activities on a wide range of micro-organisms, while bombinin H only exhibited a mildly bacteriostatic effect on the Gram-positive bacteria Staphylococcus aureus . The combination potency of BHL-bombinin with either bombinin HL or bombinin HD showed the synergistic inhibition activities against S. aureus (FICI 0.375). A synergistic effect has also been observed between bombinin H and ampicillin, which was further systematically evaluated and confirmed by in vitro time-killing investigations. Haemolytic and cytotoxic examinations exhibited a highly synergistic selectivity and low cytotoxicity on mammalian cells of these three peptides. Taken together, the discovery of the potent synergistic effect of AMPs in a single biosynthetic precursor with superior functional selectivity provides a promising strategy to combat multidrug-resistant pathogens in clinical therapy
Original languageEnglish
JournalBioscience Reports
DOIs
Publication statusPublished - 11 Sep 2017

Keywords

  • antimicrobial; peptide; selectivity; skin secretion; synergism

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