Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2017 Nov 7;22(11):1896.
doi: 10.3390/molecules22111896.

PSN-PC: A Novel Antimicrobial and Anti-Biofilm Peptide from the Skin Secretion of Phyllomedusa-camba with Cytotoxicity on Human Lung Cancer Cell

Xianhui Wu  1   2 , Jinhuo Pan  3 , Yue Wu  4 , Xinping Xi  5 , Chengbang Ma  6 , Lei Wang  7 , Mei Zhou  8 , Tianbao Chen  9
Affiliations

PSN-PC: A Novel Antimicrobial and Anti-Biofilm Peptide from the Skin Secretion of Phyllomedusa-camba with Cytotoxicity on Human Lung Cancer Cell

Xianhui Wu et al. Molecules. .

Abstract

Peptides derived from amphibian skin secretion are promising drug prototypes for combating widespread infection. In this study, a novel peptide belonging to the phylloseptin family of antimicrobial peptides was isolated from the skin secretion of the Phyllomedusa camba, namely phylloseptin-PC (PSN-PC). The biosynthetic precursor was obtained by molecular cloning and the mature peptide sequence was confirmed through tandem mass spectrometry (MS/MS) fragmentation sequencing in the skin secretion. The synthetic replicate exhibited a broad spectrum antimicrobial activity against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus,Escherichia coli, Pseudomonas aeruginosa, Candida albicans at concentrations of 2, 2, 8, 32 and 2 µM, respectively. It also showed the capability of eliminating S. aureus biofilm with a minimal biofilm eradication concentration of 8 µM. The haemolysis of this peptide was not significant at low concentrations but had a considerable increase at high concentrations. Additionally, this peptide showed an anti-proliferation effect on the non-small cell lung cancer cell line (NCI-H157), with low cytotoxicity on the human microvascular endothelial cell line (HMEC-1). The discovery of the novel peptide may provide useful clues for new drug discoveries.

Keywords: anti-biofilm activity; antimicrobial peptide; cancer cell cytotoxicity; phylloseptin.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
Nucleotide and translated open-reading frame amino acid sequence of biosynthetic precursor cDNA encoding the novel mature peptide. The putative signal peptide is single-underlined, the mature peptide is double-underlined, and the stop codon is indicated by an asterisk.
Figure 2
Figure 2
Multiple alignments of the cloned cDNA-deduced amino acid sequence of phylloseptins with antimicrobial activities. Grey shading indicates identical amino acid residues, yellow shading indicates consensus amino acid residues, and green shading indicates similar amino acid residues. (1): putative signal peptide; (2): acidic spacer peptide region; (3): dibasic propeptide convertase processing site; (4): mature peptide; (5): glycine residue amide donor.
Figure 3
Figure 3
(a) Reverse phase high performance liquid chromatography (HPLC) chromatogram of skin secretion of Phyllomedusa camba monitored at 214 nm. The arrow indicated the retention time of PSN-PC; (b) Tandem mass (MS/MS) fragmentation spectrum of PSN-PC; (c) Predicted singly-charged b ions and y ions arising from MS/MS fragmentation. The observed b- and y-ions were indicated in blue and red typefaces.
Figure 3
Figure 3
(a) Reverse phase high performance liquid chromatography (HPLC) chromatogram of skin secretion of Phyllomedusa camba monitored at 214 nm. The arrow indicated the retention time of PSN-PC; (b) Tandem mass (MS/MS) fragmentation spectrum of PSN-PC; (c) Predicted singly-charged b ions and y ions arising from MS/MS fragmentation. The observed b- and y-ions were indicated in blue and red typefaces.
Figure 4
Figure 4
(a) The RP-HPLC chromatogram of the purified synthetic phylloseptin-PC (PSN-PC); (b) MALDI-TOF mass spectrum of synthetic PSN-PC; (c) Predicted secondary structure of PSN-PC using I-TASSER; (d) Predicted 3D model of PSN-PC using I-TASSER; (e) Z-score plot using ProSA-web; (f) Helical wheel plot of PSN-PC; (g) Circular dichroism (CD) spectra recorded for PSN-PC (100 μM) in 10 mM ammonium acetate buffer and 50% TFE ammonium acetate buffer.
Figure 4
Figure 4
(a) The RP-HPLC chromatogram of the purified synthetic phylloseptin-PC (PSN-PC); (b) MALDI-TOF mass spectrum of synthetic PSN-PC; (c) Predicted secondary structure of PSN-PC using I-TASSER; (d) Predicted 3D model of PSN-PC using I-TASSER; (e) Z-score plot using ProSA-web; (f) Helical wheel plot of PSN-PC; (g) Circular dichroism (CD) spectra recorded for PSN-PC (100 μM) in 10 mM ammonium acetate buffer and 50% TFE ammonium acetate buffer.
Figure 5
Figure 5
The MBEC (minimal biofilm eradication concentration) of PSN-PC against S. aureus biofilm. The results were analysed by one-way ANOVA, followed by the Newman-Keuls test, and showed differences between growth control and all concentrations of PSN-PC (***) p < 0.001.
Figure 6
Figure 6
(a) Time-killing curves for PSN-PC on S. aureus. The antimicrobial peptide was added at time 0 h; (b) Cell-membrane permeability effects of S. aureus incubated with the peptide. The results were analysed by one-way ANOVA, followed by the Newman-Keuls test, and showed differences between 2, 4 and 8 µM (b) (***) p < 0.001.
Figure 7
Figure 7
Dose-response curves of PSN-PC on the non-small cell lung cancer cell line NCl-H157 (a) and the human microvessel endothelial cell HMEC-1 (b) after 24 h incubation. IC50 of NCl-H157 and HMEC-1 were 2.85 and 51.83 µM, respectively.
Figure 8
Figure 8
Haemolytic activity of PSN-PC. Percentage of haemolysis was calculated in comparison to the positive control using TritonX-100.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Kückelhaus S.A., Leite J.R.S., Muniz-Junqueira M.I., Sampaio R.N., Bloch C., Tosta C.E. Antiplasmodial and antileishmanial activities of phylloseptin-1, an antimicrobial peptide from the skin secretion of Phyllomedusa azurea (Amphibia) Exp. Parasitol. 2009;123:11–16. doi: 10.1016/j.exppara.2009.05.002. - DOI - PubMed
    1. Conlon J.M., Mechkarska M., Lukic M.L., Flatt P.R. Potential therapeutic applications of multifunctional host-defense peptides from frog skin as anti-cancer, anti-viral, immunomodulatory, and anti-diabetic agents. Peptides. 2014;57:67–77. doi: 10.1016/j.peptides.2014.04.019. - DOI - PubMed
    1. Zairi A., Belaïd A., Gahbiche A., Hani K. Spermicidal activity of dermaseptins. Contraception. 2005;72:447–453. doi: 10.1016/j.contraception.2005.06.055. - DOI - PubMed
    1. Ageitos J., Sánchez-Pérez A., Calo-Mata P., Villa T. Antimicrobial peptides (amps): Ancient compounds that represent novel weapons in the fight against bacteria. Biochem. Pharmacol. 2017;133:117–138. doi: 10.1016/j.bcp.2016.09.018. - DOI - PubMed
    1. Grassi L., Maisetta G., Maccari G., Esin S., Batoni G. Analogs of the frog-skin antimicrobial peptide temporin 1tb exhibit a wider spectrum of activity and a stronger antibiofilm potential as compared to the parental peptide. Front. Chem. 2017;5 doi: 10.3389/fchem.2017.00024. - DOI - PMC - PubMed

MeSH terms

Substances

LinkOut - more resources