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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.


Alignments of cDNA-deduced open-reading frame amino acid sequences of four PS precursors, PSN-9 (Accession No.Q0VZ38), PSN-1 (Accession No.Q800R3), PS-Du and PS-Co, demonstrating 7-regions of topological structures with asterisks indicating sites of amino acid sequence identities. 1—Putative signal peptide; 2 & 4—Acidic amino acid residue-rich spacer peptides; 3 & 5—Dibasic propeptide convertase processing site; 6—Mature PS peptide; 7—Glycine residue amide donor.
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toxins-08-00255-f003: Alignments of cDNA-deduced open-reading frame amino acid sequences of four PS precursors, PSN-9 (Accession No.Q0VZ38), PSN-1 (Accession No.Q800R3), PS-Du and PS-Co, demonstrating 7-regions of topological structures with asterisks indicating sites of amino acid sequence identities. 1—Putative signal peptide; 2 & 4—Acidic amino acid residue-rich spacer peptides; 3 & 5—Dibasic propeptide convertase processing site; 6—Mature PS peptide; 7—Glycine residue amide donor.

Mentions: Both putative mature peptides were subjected to bioinformatics analysis by use of the National Center for Biotechnology Information (NCBI ) protein Basic Local Alignment Search Tool (BLASTp) program, which found that PS-Du and PS-Co were new phylloseptins. PS-Du and PS-Co showed a high degree of structural identity to phylloseptins from other Phyllomedusa frogs, including the well-studied PSN-9 (accession No. Q0VZ38) from Phyllomedusa hypochondrialis and PSN-1 (accession No. Q800R3) from Phyllomedusa bicolor. The alignment of open-reading frame nucleotide and amino acid sequences of PS-DU, PS-Co, PSN-9 and PSN-1, was established by use of Vector NTI software (Version 11.5, 2010, Life Technologies, Carlsbad, CA, USA), and these are shown in Figure 2 and Figure 3. The nucleotide sequence of both PS-Du and PS-Co precursors, have been deposited in the European Molecular Biology Laboratory (EMBL) Nucleotide Sequence Database under the accession codes LN999522 and LN999523.


Discovery of Novel Bacterial Cell-Penetrating Phylloseptins in Defensive Skin Secretions of the South American Hylid Frogs, Phyllomedusa duellmani and Phyllomedusa coelestis
Alignments of cDNA-deduced open-reading frame amino acid sequences of four PS precursors, PSN-9 (Accession No.Q0VZ38), PSN-1 (Accession No.Q800R3), PS-Du and PS-Co, demonstrating 7-regions of topological structures with asterisks indicating sites of amino acid sequence identities. 1—Putative signal peptide; 2 & 4—Acidic amino acid residue-rich spacer peptides; 3 & 5—Dibasic propeptide convertase processing site; 6—Mature PS peptide; 7—Glycine residue amide donor.
© Copyright Policy
Related In: Results  -  Collection

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

toxins-08-00255-f003: Alignments of cDNA-deduced open-reading frame amino acid sequences of four PS precursors, PSN-9 (Accession No.Q0VZ38), PSN-1 (Accession No.Q800R3), PS-Du and PS-Co, demonstrating 7-regions of topological structures with asterisks indicating sites of amino acid sequence identities. 1—Putative signal peptide; 2 & 4—Acidic amino acid residue-rich spacer peptides; 3 & 5—Dibasic propeptide convertase processing site; 6—Mature PS peptide; 7—Glycine residue amide donor.
Mentions: Both putative mature peptides were subjected to bioinformatics analysis by use of the National Center for Biotechnology Information (NCBI ) protein Basic Local Alignment Search Tool (BLASTp) program, which found that PS-Du and PS-Co were new phylloseptins. PS-Du and PS-Co showed a high degree of structural identity to phylloseptins from other Phyllomedusa frogs, including the well-studied PSN-9 (accession No. Q0VZ38) from Phyllomedusa hypochondrialis and PSN-1 (accession No. Q800R3) from Phyllomedusa bicolor. The alignment of open-reading frame nucleotide and amino acid sequences of PS-DU, PS-Co, PSN-9 and PSN-1, was established by use of Vector NTI software (Version 11.5, 2010, Life Technologies, Carlsbad, CA, USA), and these are shown in Figure 2 and Figure 3. The nucleotide sequence of both PS-Du and PS-Co precursors, have been deposited in the European Molecular Biology Laboratory (EMBL) Nucleotide Sequence Database under the accession codes LN999522 and LN999523.

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.