The Effect of the Lysine Acetylation Modification of ClpP on the Virulence of Vibrio alginolyticus
Abstract
:1. Introduction
2. Results
2.1. Characteristics of clpP
2.2. Recombinant ClpP Expression, Purification and Antibody Validation
2.3. Validation of ClpP Lysine-Acetylated Proteins Using Immunoprecipitation and Western Blot
2.4. Growth Curve Measurement
2.5. Effect of Lysine Acetylation on Biofilm Formation
2.6. Effect of Site-Directed Mutagenesis Strains on LD50 of Zebrafish
2.7. Effect of Site-Directed Mutagenesis Strains on ECPase Activity
2.8. Effects of Site-Directed Mutagenesis Strains on GS Cells
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Fish
4.2. Cloning and Bioinformatics Analysis of clpP Gene from V. alginolyticus HY9901
4.3. Prokaryotic Expression of clpP and Antibody Preparation
4.4. Immunoprecipitation and Western Blot
4.5. Construction of clpP Complementation Strain (C-clpP) and Acetylation Site-Directed Mutagenesis Strains
4.6. Growth Curve of Bacteria
4.7. Quantification of Biofilm Biomass
4.8. Fifty Percent Lethal Dose (LD50)
4.9. Detection of Extracellular Protease (ECPase) Activity
4.10. GS Cell Cytotoxicity Assay
4.11. Statistical Analysis
5. 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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Strain | QMEAN | Cβ | All Atom | Solvation | Torsion |
---|---|---|---|---|---|
ClpP | 1.14 | 1.03 | 1.03 | 1.88 | 0.32 |
K165Q | 1.26 | 0.98 | 1.10 | 1.88 | 0.44 |
K165R | 1.14 | 1.03 | 1.01 | 1.93 | 0.31 |
WT | C-clpP | K165Q | K165R | |
---|---|---|---|---|
LD50 (cfu/mL) | 5.82 × 106 | 5.89 × 106 | 9.75 × 107 ** | 7.34 × 106 |
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Wang, S.; Jiang, Y.; Zhang, W.; Wei, Y.; Xiao, X.; Wei, Z.; Wen, X.; Dong, Y.; Jian, J.; Wang, N.; et al. The Effect of the Lysine Acetylation Modification of ClpP on the Virulence of Vibrio alginolyticus. Molecules 2024, 29, 4278. https://doi.org/10.3390/molecules29174278
Wang S, Jiang Y, Zhang W, Wei Y, Xiao X, Wei Z, Wen X, Dong Y, Jian J, Wang N, et al. The Effect of the Lysine Acetylation Modification of ClpP on the Virulence of Vibrio alginolyticus. Molecules. 2024; 29(17):4278. https://doi.org/10.3390/molecules29174278
Chicago/Turabian StyleWang, Shi, Yingying Jiang, Weijie Zhang, Yingzhu Wei, Xing Xiao, Zhiqing Wei, Xiaoxin Wen, Yuhang Dong, Jichang Jian, Na Wang, and et al. 2024. "The Effect of the Lysine Acetylation Modification of ClpP on the Virulence of Vibrio alginolyticus" Molecules 29, no. 17: 4278. https://doi.org/10.3390/molecules29174278