Novel Antimicrobial Peptides from a Cecropin-Like Region of Heteroscorpine-1 from Heterometrus laoticus Venom with Membrane Disruption Activity
Abstract
:1. Introduction
2. Results
2.1. Peptides Design and Physicochemical Properties
2.2. Structural Analysis of Peptides
2.3. Antimicrobial Activity
2.4. Synergistic Activity Assay
2.5. Time-Killing Assay
2.6. Outer Membrane Permeability Assay
2.7. Inner Membrane Permeability Assay
2.8. Scanning Electron Microscope (SEM) Analysis
2.9. Hemolytic Activity Assay
2.10. Cytotoxic Assay
3. Discussion
4. Materials and Methods
4.1. Peptides Design
4.2. Peptides Synthesis
4.3. CD Spectroscopy
4.4. Antimicrobial Activity
4.5. Synergistic Activity Assay
4.6. Time Killing Assay
4.7. Outer Membrane Permeability Assay
4.8. Inner Membrane Permeability Assay
4.9. Scanning Electron Microscope
4.10. Hemolytic Activity Assay
4.11. Cytotoxic Assay
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Peptides | UniProtKB ID | Origin | Sequence | AA | Identity (%) | Reference |
---|---|---|---|---|---|---|
CeHS-1 | - | Heterometrus laoticus | GWINEEKIQKKIDEKIGNNILGGMA KA--VVHKLAKGEFQ-- | 38 | - | This work |
Cecropin B | AP00128 | Antheraea pernyi | KW----KIFKKI-EKVGRNIRNGII KAGPAVAVL--GEAKAL | 35 | 42.85 | [30] |
CM4 | AP01259 | Bombyx mori | RWKIFKKI-----EKVGQNIRDGIV KAGPAVAVVGQAATI | 35 | 41.86 | [33] |
Hyphancin IIIF | AP00348 | Hyphantria cunea | RWKVFKKI-----EKVGRNIRDGVI KAGPAIAVVGQAKAL | 35 | 41.86 | [34] |
Peptides | Molecular Weight (MW) | Charge | %H | H | μH | α-Helix | |
---|---|---|---|---|---|---|---|
Theoretical | Observed | ||||||
CeHS-1 | 4235.93 | 4234.8 | +2 | 39.47 | 0.228 | 0.110 | 76.32% |
CeHS-1 GP | 4016.78 | 4016.0 | +5 | 43.24 | 0.309 | 0.463 | 70.27% |
CeHS-1 GPK | 4030.85 | 4029.5 | +6 | 43.24 | 0.299 | 0.468 | 67.57% |
Peptides | % of Content in Water | % of Content in 40% TFE | ||
---|---|---|---|---|
α-Helix | β-Sheet | α-Helix | β-Sheet | |
CeHS-1 | 8.02 | 22.14 | 8.02 | 22.14 |
CeHS-1 GP | 62.6 | 3.79 | 87.59 | 0.48 |
CeHS-1 GPK | 62.6 | 3.79 | 87.59 | 0.48 |
Bacterial Strain | CeHS-1 | CeHS-1 GP | CeHS-1 GPK | Ampicillin | ||||
---|---|---|---|---|---|---|---|---|
MIC | MBC | MIC | MBC | MIC | MBC | MIC | MBC | |
K. pneumoniae ATCC 27853 | 64 | >128 | 32 | 64 | 32 | 128 | 0.06 | 0.13 |
E. coli ATCC 25922 | 64 | >128 | 16 | 16 | 8 | 8 | 1 | 1 |
B. subtilis TISR 008 | 64 | 64 | 32 | 64 | 8 | 16 | 0.06 | 0.06 |
S. aureus ATCC 25923 | 64 | >128 | 32 | 128 | 16 | 64 | 0.06 | 0.25 |
P. aeruginosa ATCC 27853 | 64 | 64 | 32 | 32 | 16 | 32 | 0.13 | 0.25 |
resistant K. pneumoniae | 16 | >128 | 32 | >128 | 32 | >128 | >128 | >128 |
MRSA | 128 | >128 | 128 | >128 | 128 | >128 | 32 | 32 |
Bacterial Strain | FIC in Combination with | |
---|---|---|
Ampicillin | Kanamycin | |
E. coli ATCC 25922 | 0.75 | 1 |
B. subtilis TISR 008 | 0.75 | 0.5 |
S. aureus ATCC 25923 | 3 | 2 |
P. aeruginosa ATCC 27853 | 0.75 | 0.5 |
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Erviana, R.; Saengkun, Y.; Rungsa, P.; Jangpromma, N.; Tippayawat, P.; Klaynongsruang, S.; Daduang, J.; Daduang, S. Novel Antimicrobial Peptides from a Cecropin-Like Region of Heteroscorpine-1 from Heterometrus laoticus Venom with Membrane Disruption Activity. Molecules 2021, 26, 5872. https://0-doi-org.brum.beds.ac.uk/10.3390/molecules26195872
Erviana R, Saengkun Y, Rungsa P, Jangpromma N, Tippayawat P, Klaynongsruang S, Daduang J, Daduang S. Novel Antimicrobial Peptides from a Cecropin-Like Region of Heteroscorpine-1 from Heterometrus laoticus Venom with Membrane Disruption Activity. Molecules. 2021; 26(19):5872. https://0-doi-org.brum.beds.ac.uk/10.3390/molecules26195872
Chicago/Turabian StyleErviana, Rima, Yutthakan Saengkun, Prapenpuksiri Rungsa, Nisachon Jangpromma, Patcharaporn Tippayawat, Sompong Klaynongsruang, Jureerut Daduang, and Sakda Daduang. 2021. "Novel Antimicrobial Peptides from a Cecropin-Like Region of Heteroscorpine-1 from Heterometrus laoticus Venom with Membrane Disruption Activity" Molecules 26, no. 19: 5872. https://0-doi-org.brum.beds.ac.uk/10.3390/molecules26195872