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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

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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.


Nucleotide and translated open-reading frame amino acid sequences of cloned cDNAs encoding precursors of novel phylloseptin peptides, PS-Du (A) and PS-Co (B). Putative signal peptides are double-underlined, mature peptides are single–underlined and stop codons are indicated by asterisks.
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toxins-08-00255-f001: Nucleotide and translated open-reading frame amino acid sequences of cloned cDNAs encoding precursors of novel phylloseptin peptides, PS-Du (A) and PS-Co (B). Putative signal peptides are double-underlined, mature peptides are single–underlined and stop codons are indicated by asterisks.

Mentions: Degenerate primers were used for interrogating the skin secretion-derived cDNA libraries of Phyllomedusa duellmani and Phyllomedusa coelestis. Two full-length cDNAs, encoding PS-Du and PS-Co, were cloned repeatedly (at least 10 clones for each) from the skin secretion-derived cDNA libraries of Phyllomedusa duellmani and Phyllomedusa coelestis, respectively. They were named PS-Du and PS-Co, respectively, reflecting their species names. The sequences of nucleotides and translated open reading frame amino acids for both peptides are shown in Figure 1A, B. Their structural topology consisted of five typical regions, including a putative signal peptide region of 22 amino acid residues, an acidic “spacer” peptide, typical -KR- propeptide convertase processing sites, a mature peptide of 19 amino acid residues and a Gly residue at the C-terminus which acts as an amide donor for providing the post-translational amide modification in each case.


Discovery of Novel Bacterial Cell-Penetrating Phylloseptins in Defensive Skin Secretions of the South American Hylid Frogs, Phyllomedusa duellmani and Phyllomedusa coelestis
Nucleotide and translated open-reading frame amino acid sequences of cloned cDNAs encoding precursors of novel phylloseptin peptides, PS-Du (A) and PS-Co (B). Putative signal peptides are double-underlined, mature peptides are single–underlined and stop codons are indicated by asterisks.
© Copyright Policy
Related In: Results  -  Collection

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

toxins-08-00255-f001: Nucleotide and translated open-reading frame amino acid sequences of cloned cDNAs encoding precursors of novel phylloseptin peptides, PS-Du (A) and PS-Co (B). Putative signal peptides are double-underlined, mature peptides are single–underlined and stop codons are indicated by asterisks.
Mentions: Degenerate primers were used for interrogating the skin secretion-derived cDNA libraries of Phyllomedusa duellmani and Phyllomedusa coelestis. Two full-length cDNAs, encoding PS-Du and PS-Co, were cloned repeatedly (at least 10 clones for each) from the skin secretion-derived cDNA libraries of Phyllomedusa duellmani and Phyllomedusa coelestis, respectively. They were named PS-Du and PS-Co, respectively, reflecting their species names. The sequences of nucleotides and translated open reading frame amino acids for both peptides are shown in Figure 1A, B. Their structural topology consisted of five typical regions, including a putative signal peptide region of 22 amino acid residues, an acidic “spacer” peptide, typical -KR- propeptide convertase processing sites, a mature peptide of 19 amino acid residues and a Gly residue at the C-terminus which acts as an amide donor for providing the post-translational amide modification in each case.

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.