Myxinidin-Derived Peptide against Biofilms Caused by Cystic Fibrosis Emerging Pathogens
Abstract
:1. Introduction
2. Results
2.1. WMR Serum Stability and Design of Novel Analogues
2.2. Minimal Inhibitory Concentration (MIC)
2.3. Serum Stability of the Analogue WMR-4
2.4. Biofilm Formation Assay and Antibiofilm Activity
2.5. RT-PCR
2.6. In Vivo Toxicity and Activity of the Peptide on the Infected G. mellonella
2.7. Peptide Secondary Structure
2.8. Interaction of WMR-4 with Lipopolysaccharide (LPS)
2.9. WMR-4: Liposome Insertion and Leakage
3. Materials and Methods
3.1. Peptide Synthesis
3.2. Strains and Cultural Conditions
3.3. Minimum Inhibitory Concentration (MIC)
3.4. Proteolitic Stability
3.5. Biofilm Formation Assay
3.6. Antibiofilm Activity
3.7. Quantitative Real Time PCR
3.8. In Vivo Effect of WMR-4 on Galleria mellonella Larvae
3.9. Statistical Analyses
3.10. Circular Dichroism (CD) Spectroscopy
3.11. Liposome Preparations
3.12. Peptide Aggregation by Thioflavin T Assay
3.13. Liposomes Leakage by ANT/DPX Assay
3.14. Tryptophan Quenching by Acrylamide
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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MW (Da) | Fragment * |
---|---|
1760.15 | NH2-WGIRRILKYGKRSK-CONH2 |
1443.86 | NH2-WGIRRILKYGKRS(K)-CONH2 |
1543.86 | NH2-WGIRRILKYGKR(SK)-CONH2 |
1361.80 | NH2-(W)GIRRILKYGKRSK-CONH2 |
Name | Sequence |
---|---|
WMR | NH2-WGIRRILKYGKRSK-CONH2 |
WMR-1 | NH2-WGIRRILKYGKRSk-CONH2 |
WMR-2 | NH2-WGIRRILKYGKRsK-CONH2 |
WMR-3 | NH2-WAibIRRILKYGKRSKCONH2 |
WMR-4 | NH2-WAibIRRILKYGKRskCONH2 |
WMR-5 | NH2-WGIRRILKYGKRSDapCONH2 |
MIC (mM) | ||||||||
---|---|---|---|---|---|---|---|---|
WMR | WMR-1 | WMR-2 | WMR-3 | WMR-4 | WMR-5 | FLC (µg/mL) | MEM (µg/mL) | |
C. albicans ATCC 90028 | >50 | >50 | >50 | >50 | 50 | >50 | 16 (48) | - |
A. xylosoxidans DSM 2402 | 50 | >50 | >50 | >50 | 25 | 25 | - | 65 (250) |
S. maltophilia DSM 50170 | 50 | >50 | >50 | >50 | 25 | 25 | - | 130 (499) |
Micro-Organism | Primer | Primer Sequence (5′->3′) |
---|---|---|
A. xylosoxidans | Usp-A |
F: CCACGAAGATCCGTACCAGG R: AGGCCTTCTTGCGGTACAC |
AxyA | F: CTGGAAGACGGGTCGCAATA R: TGAGTTCACGCAGTTCCACT | |
espF | F: GCGTTCATCTATCCGGCCAT R: TGATCTCCATGATCGGCAGC | |
rRNA16S | F: GCAGCAGTGGGGAATTTTGG R: ATTTCACTGCTACACGCGGA | |
S. maltophilia | smf-1 | F: GGAAGGTATGTCCGAGTCCG R: GCGGGTACGGCTACGATCAGTT |
Ax21 | F: GGCTACAACGTCGAAATCGC R: ATTCTTCAGCTCGCCGTTCA | |
xanB | F: TATGCGATCGATGCGTCCAA R: CGAATGCGATCTCTTCGGGA | |
rpfF | F: CTGGTCGACATCGTGGTG R: TGATCCGCATCATTTCATGC | |
rRNA 16s | F: ACTGAGACACGGTCCAGACT R: CTTCTTCACCCACGCGGTAT | |
C. albicans | ERG11 | F: ATTGTTGAAACTGTCATTG R: CCCCTAATAATATACTGATCTG |
HOG1 | F: GACTTGTGGTCTGTGGGTTG R: ACATCAGCAGGAGGTGAGC | |
ALS3 | F: CTAATGCTGCTACGTATAATT R: CCTGAAATTGACATGTAGCA | |
ACT | F: AGCCCAATCCAAAAGAGGTATT R: GCTTCGGTCAACAAAACTGG |
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Bellavita, R.; Maione, A.; Braccia, S.; Sinoca, M.; Galdiero, S.; Galdiero, E.; Falanga, A. Myxinidin-Derived Peptide against Biofilms Caused by Cystic Fibrosis Emerging Pathogens. Int. J. Mol. Sci. 2023, 24, 3092. https://doi.org/10.3390/ijms24043092
Bellavita R, Maione A, Braccia S, Sinoca M, Galdiero S, Galdiero E, Falanga A. Myxinidin-Derived Peptide against Biofilms Caused by Cystic Fibrosis Emerging Pathogens. International Journal of Molecular Sciences. 2023; 24(4):3092. https://doi.org/10.3390/ijms24043092
Chicago/Turabian StyleBellavita, Rosa, Angela Maione, Simone Braccia, Marica Sinoca, Stefania Galdiero, Emilia Galdiero, and Annarita Falanga. 2023. "Myxinidin-Derived Peptide against Biofilms Caused by Cystic Fibrosis Emerging Pathogens" International Journal of Molecular Sciences 24, no. 4: 3092. https://doi.org/10.3390/ijms24043092