A Novel Dermaseptin Isolated from the Skin Secretion of Phyllomedusa tarsius and Its Cationicity-Enhanced Analogue Exhibiting Effective Antimicrobial and Anti-Proliferative Activities
Abstract
:1. Introduction
2. Materials and Methods
2.1. Acquisition of Phyllomedusa tarsius Skin Secretion
2.2. “Shotgun” Cloning of DPT9 Precursor-Encoding cDNA from a Skin Secretion-Derived cDNA Library
2.3. Isolation and Identification of DPT9 from Crude Skin Secretion
2.4. Chemical Synthesis of DPT9 and K8, 23-DTP9
2.5. Secondary Structure Analysis of DPT9 and K8, 23-DTP9
2.6. Antimicrobial and Antibiofilm Susceptibility Assay
2.7. Haemolysis Assay
2.8. Membrane Permeability Assay
2.9. MTT Assay
2.10. Lactate Dehydrogenase (LDH) Assay
3. Results
3.1. “Shotgun” Cloning of DPT9 Biosynthetic Precursor cDNA from a Skin Secretion-Derived cDNA Library of Phyllomedusa tarsius
3.2. Identification and Structural Characterization of DPT9
3.3. Synthesis, Predicted Physicochemical Properties and Secondary Structure of DPT9 and its Analogue K8, 23-DTP9
3.4. Antimicrobial and Haemolytic Activities of DPT9 and its Analogue K8, 23-DTP9
3.5. Membrane Permeabilisation of DPT9 and Its Analogue K8, 23-DTP9
3.6. MTT and LDH Assays on the Human Cancer and Normal Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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DPT9 | K8, 23-DPT9 | DPH a | DCA1 b | DDU1 b | DPS3 c | DPS4 d | |
---|---|---|---|---|---|---|---|
Molecular weight (Da) | 2605.10 | 2617.24 | 2535.78 | 3024.62 | 2911.46 | 2550.00 | 2948.53 |
Hydrophobicity | 0.348 | 0.326 | 0.333 | 0.291 | 0.331 | 0.373 | 0.368 |
Hydrophobic moment | 0.447 | 0.462 | 0.427 | 0.263 | 0.255 | 0.437 | 0.226 |
Net charge | +2 | +6 | +1 | +3 | +3 | +2 | +3 |
α-helix (%) | 24.18 | 46.14 | 35 | 25.3 | 27.9 | 44.9 | 67.6 |
Microorganisms | MIC/MBC (µM) | |||||||
---|---|---|---|---|---|---|---|---|
MIC/MBC (µg/mL) | ||||||||
DPT9 | K8, 23-DPT9 | DPH | DCA1 | DDU1 | DPS3 a | DPS4 a | Melittin | |
S. aureus NCTC 10788 | 16/32 | 2/4 | 32/64 | 4/16 | 4/16 | 256 | 4 | 1/2 |
41.7/83.4 | 5.2/10.4 | 81.1/162.2 | 12.1/48.4 | 11.6/46.4 | 652.5 | 11.8 | 2.8/5.7 | |
Methicillin-resistant Staphylococcus aureus (MRSA) NCTC 12493 | 32/64 | 2/4 | >512/>512 | 8/32 | 4/16 | NA | 8 | 1/4 |
83.4/166.7 | 5.2/10.4 | >1298.4/>1298.4 | 24.2/96.8 | 11.6/46.4 | 23.6 | 2.8/11.4 | ||
E. faecalis NCTC 12697 | 16/32 | 2/4 | NA | 128/256 | 64/128 | NA | 32 | 1/2 |
41.7/83.4 | 5.2/10.4 | 387.1/774.1 | 185.6/371.2 | 94.3 | 2.8/5.7 | |||
E. coli NCTC 10418 | 8/16 | 2/4 | 16/16 | 4/16 | 4/16 | 8 | 8 | 2/4 |
20.8/41.7 | 5.2/10.4 | 40.6/40.6 | 12.1/48.4 | 11.6/46.4 | 20.4 | 23.6 | 5.7/11.4 | |
P. aeruginosa ATCC 27853 | 16/32 | 4/8 | 64>512 | 8/32 | 4/16 | NA | 16 | 16/32 |
41.7/83.4 | 10.4/20.8 | 162.2/>1298.4 | 24.2/96.8 | 11.6/46.4 | 47.1 | 45.5/91.1 | ||
K. pneumoniae ATCC 43816 | 8/16 | 2/4 | NA | 8/128 | 4/64 | NA | NA | 2/8 |
20.8/41.7 | 5.2/10.4 | 24.2/387.1 | 11.6/185.6 | 5.7/22.8 | ||||
C. albicans NCYC 1467 | 64/128 | 4/8 | 16/64 | 4/16 | 4/16 | 4 | 4 | 2/4 |
166.7/333.4 | 10.4/20.8 | 40.6/162.2 | 12.1/48.4 | 11.6/46.4 | 10.2 | 11.8 | 5.7/11.4 |
Microorganisms | MBIC/MBEC (μM) a | |
---|---|---|
DPT9 | K8, 23-DPT9 | |
S. aureus NCTC 10788 | 16/32 | 2/4 |
MRSA NCTC 12493 | 32/64 | 2/4 |
E. coli NCTC 10418 | 8/16 | 2/4 |
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Li, M.; Xi, X.; Ma, C.; Chen, X.; Zhou, M.; Burrows, J.F.; Chen, T.; Wang, L. A Novel Dermaseptin Isolated from the Skin Secretion of Phyllomedusa tarsius and Its Cationicity-Enhanced Analogue Exhibiting Effective Antimicrobial and Anti-Proliferative Activities. Biomolecules 2019, 9, 628. https://doi.org/10.3390/biom9100628
Li M, Xi X, Ma C, Chen X, Zhou M, Burrows JF, Chen T, Wang L. A Novel Dermaseptin Isolated from the Skin Secretion of Phyllomedusa tarsius and Its Cationicity-Enhanced Analogue Exhibiting Effective Antimicrobial and Anti-Proliferative Activities. Biomolecules. 2019; 9(10):628. https://doi.org/10.3390/biom9100628
Chicago/Turabian StyleLi, Miaoran, Xinping Xi, Chengbang Ma, Xiaoling Chen, Mei Zhou, James F. Burrows, Tianbao Chen, and Lei Wang. 2019. "A Novel Dermaseptin Isolated from the Skin Secretion of Phyllomedusa tarsius and Its Cationicity-Enhanced Analogue Exhibiting Effective Antimicrobial and Anti-Proliferative Activities" Biomolecules 9, no. 10: 628. https://doi.org/10.3390/biom9100628