A Rapid Fluorescence-Based Microplate Assay to Investigate the Interaction of Membrane Active Antimicrobial Peptides with Whole Gram-Positive Bacteria
Abstract
:1. Introduction
2. Results
2.1. Compatibility between the Fluorescent Dyes
2.2. Selection of Membrane-Active Peptides
2.3. Interference of Peptides and Uncouplers with the Fluorescent Dyes
2.4. Monitoring of Peptide-Induced Membrane Alterations with the Combination of diSC3(5) and PI
2.5. Morphology of Peptide-Treated S. epidermidis
3. Discussion
4. Materials and Methods
4.1. Equipment
4.2. Peptides and Other Reagents
4.3. Bacteria and Bacterial Cultures
4.4. Determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
4.5. Excitation and Emission Spectra
4.6. Interference of Uncouplers and Peptides with the Fluorescent Dyes
4.7. Kinetic Fluorescence Measurements to Detect Membrane Depolarization and Permeabilization
4.8. Field Emission Scanning Electron Microscopy (FE-SEM) of S. epidermidis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cecropin A | Melittin | Magainin 2 | Gramicidin D | |
---|---|---|---|---|
MIC (MBC) (µM) a,b | ||||
S. epidermidis ATCC 35984 | >128 | 0.5 (0.5) | >128 | 2 (4) |
S. aureus ATCC 25923 | >128 | 0.5 (1) | 64 (128) | 4 (8) |
E. coli ATCC 25922 | 0.75 (0.75) | 1 (1) | 8 (16) | >32 |
P. aeruginosa ATCC 27853 | 1 (2) | 1 (2) | 64 (64) | >32 |
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Boix-Lemonche, G.; Lekka, M.; Skerlavaj, B. A Rapid Fluorescence-Based Microplate Assay to Investigate the Interaction of Membrane Active Antimicrobial Peptides with Whole Gram-Positive Bacteria. Antibiotics 2020, 9, 92. https://doi.org/10.3390/antibiotics9020092
Boix-Lemonche G, Lekka M, Skerlavaj B. A Rapid Fluorescence-Based Microplate Assay to Investigate the Interaction of Membrane Active Antimicrobial Peptides with Whole Gram-Positive Bacteria. Antibiotics. 2020; 9(2):92. https://doi.org/10.3390/antibiotics9020092
Chicago/Turabian StyleBoix-Lemonche, Gerard, Maria Lekka, and Barbara Skerlavaj. 2020. "A Rapid Fluorescence-Based Microplate Assay to Investigate the Interaction of Membrane Active Antimicrobial Peptides with Whole Gram-Positive Bacteria" Antibiotics 9, no. 2: 92. https://doi.org/10.3390/antibiotics9020092