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Discovery of Novel Bacterial Cell-Penetrating Phylloseptins in Defensive Skin Secretions of the South American Hylid Frogs, Phyllomedusa duellmani and Phyllomedusa coelestis

View Article: PubMed Central - PubMed

ABSTRACT

Phylloseptin (PS) peptides, derived from South American hylid frogs (subfamily Phyllomedusinae), have been found to have broad-spectrum antimicrobial activities and relatively low haemolytic activities. Although PS peptides have been identified from several well-known and widely-distributed species of the Phyllomedusinae, there remains merit in their study in additional, more obscure and specialised members of this taxon. Here, we report the discovery of two novel PS peptides, named PS-Du and PS-Co, which were respectively identified for the first time and isolated from the skin secretions of Phyllomedusa duellmani and Phyllomedusa coelestis. Their encoding cDNAs were cloned, from which it was possible to deduce the entire primary structures of their biosynthetic precursors. Reversed-phase high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS) analyses were employed to isolate and structurally-characterise respective encoded PS peptides from skin secretions. The peptides had molecular masses of 2049.7 Da (PS-Du) and 1972.8 Da (PS-Co). They shared typical N-terminal sequences and C-terminal amidation with other known phylloseptins. The two peptides exhibited growth inhibitory activity against E. coli (NCTC 10418), as a standard Gram-negative bacterium, S. aureus (NCTC 10788), as a standard Gram-positive bacterium and C. albicans (NCPF 1467), as a standard pathogenic yeast, all as planktonic cultures. Moreover, both peptides demonstrated the capability of eliminating S. aureus biofilm.

No MeSH data available.


Related in: MedlinePlus

Cell-membrane permeability effects of PS-Du (A) and PS-Co (B) on S. aureus detected by the SYTOX Green (Life technologies, Carlsbad, CA, USA) assay at peptide concentrations corresponding to 1 MIC, 2 MIC and 4 MIC. Positive membrane permeabilization was obtained following incubation of S. aureus with 70% isopropyl alcohol. The negative control was represented as vehicle only. Data represent means ± SEM of 5 replicates.
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toxins-08-00255-f012: Cell-membrane permeability effects of PS-Du (A) and PS-Co (B) on S. aureus detected by the SYTOX Green (Life technologies, Carlsbad, CA, USA) assay at peptide concentrations corresponding to 1 MIC, 2 MIC and 4 MIC. Positive membrane permeabilization was obtained following incubation of S. aureus with 70% isopropyl alcohol. The negative control was represented as vehicle only. Data represent means ± SEM of 5 replicates.

Mentions: The activities of PS-Du and PS-Co against S. aureus biofilm were tested and both peptides possessed biofilm eradication capability with an MBEC (minimal biofilm eradication concentration) of 16 mg/L (Figure 11). Additionally, PS-Du and PS-Co had the capability of cell-membrane permeabilization at concentrations of 8 mg/L and 16 mg/L, respectively (Figure 12). Each assay was carried out over at least three individual experiments with three replicates in each.


Discovery of Novel Bacterial Cell-Penetrating Phylloseptins in Defensive Skin Secretions of the South American Hylid Frogs, Phyllomedusa duellmani and Phyllomedusa coelestis
Cell-membrane permeability effects of PS-Du (A) and PS-Co (B) on S. aureus detected by the SYTOX Green (Life technologies, Carlsbad, CA, USA) assay at peptide concentrations corresponding to 1 MIC, 2 MIC and 4 MIC. Positive membrane permeabilization was obtained following incubation of S. aureus with 70% isopropyl alcohol. The negative control was represented as vehicle only. Data represent means ± SEM of 5 replicates.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037481&req=5

toxins-08-00255-f012: Cell-membrane permeability effects of PS-Du (A) and PS-Co (B) on S. aureus detected by the SYTOX Green (Life technologies, Carlsbad, CA, USA) assay at peptide concentrations corresponding to 1 MIC, 2 MIC and 4 MIC. Positive membrane permeabilization was obtained following incubation of S. aureus with 70% isopropyl alcohol. The negative control was represented as vehicle only. Data represent means ± SEM of 5 replicates.
Mentions: The activities of PS-Du and PS-Co against S. aureus biofilm were tested and both peptides possessed biofilm eradication capability with an MBEC (minimal biofilm eradication concentration) of 16 mg/L (Figure 11). Additionally, PS-Du and PS-Co had the capability of cell-membrane permeabilization at concentrations of 8 mg/L and 16 mg/L, respectively (Figure 12). Each assay was carried out over at least three individual experiments with three replicates in each.

View Article: PubMed Central - PubMed

ABSTRACT

Phylloseptin (PS) peptides, derived from South American hylid frogs (subfamily Phyllomedusinae), have been found to have broad-spectrum antimicrobial activities and relatively low haemolytic activities. Although PS peptides have been identified from several well-known and widely-distributed species of the Phyllomedusinae, there remains merit in their study in additional, more obscure and specialised members of this taxon. Here, we report the discovery of two novel PS peptides, named PS-Du and PS-Co, which were respectively identified for the first time and isolated from the skin secretions of Phyllomedusa duellmani and Phyllomedusa coelestis. Their encoding cDNAs were cloned, from which it was possible to deduce the entire primary structures of their biosynthetic precursors. Reversed-phase high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS) analyses were employed to isolate and structurally-characterise respective encoded PS peptides from skin secretions. The peptides had molecular masses of 2049.7 Da (PS-Du) and 1972.8 Da (PS-Co). They shared typical N-terminal sequences and C-terminal amidation with other known phylloseptins. The two peptides exhibited growth inhibitory activity against E. coli (NCTC 10418), as a standard Gram-negative bacterium, S. aureus (NCTC 10788), as a standard Gram-positive bacterium and C. albicans (NCPF 1467), as a standard pathogenic yeast, all as planktonic cultures. Moreover, both peptides demonstrated the capability of eliminating S. aureus biofilm.

No MeSH data available.


Related in: MedlinePlus