Sphingomyelin Depletion from Plasma Membranes of Human Airway Epithelial Cells Completely Abrogates the Deleterious Actions of S. aureus Alpha-Toxin
Abstract
:1. Introduction
2. Results
2.1. Pre-Treatment of Airway Cells with rHlb Allows rHla Monomer Binding to the PM, but Prevents Formation of Heptamers
2.2. Effects of Sphingomyelinase Pre-Treatment of Airway Epithelial Cells on rHla-Mediated Changes in [Ca2+]i
2.3. Effects of Sphingomyelinase Pre-Treatment of Airway Epithelial Cells on rHla-Mediated Formation of Paracellular Gaps
2.4. Effects of Pre-Treatment of Airway Epithelial Cells with Sphingomyelinase (rHlb) on rHla-Mediated Hypo-Phosphorylation of Cofilin
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Expression and Purification of Recombinant Staphylococcus aureus Hla and Hlb
4.3. Cell Culture
4.4. Freshly Prepared Human Airway Tissue
4.5. Sample Preparation for Western Blotting
4.6. Semi-Quantitative Western Blotting
4.7. Intracellular Calcium Concentrations
4.8. Time Lapse Microscopy
4.9. Data Presentation and Statistics
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Ziesemer, S.; Möller, N.; Nitsch, A.; Müller, C.; Beule, A.G.; Hildebrandt, J.-P. Sphingomyelin Depletion from Plasma Membranes of Human Airway Epithelial Cells Completely Abrogates the Deleterious Actions of S. aureus Alpha-Toxin. Toxins 2019, 11, 126. https://doi.org/10.3390/toxins11020126
Ziesemer S, Möller N, Nitsch A, Müller C, Beule AG, Hildebrandt J-P. Sphingomyelin Depletion from Plasma Membranes of Human Airway Epithelial Cells Completely Abrogates the Deleterious Actions of S. aureus Alpha-Toxin. Toxins. 2019; 11(2):126. https://doi.org/10.3390/toxins11020126
Chicago/Turabian StyleZiesemer, Sabine, Nils Möller, Andreas Nitsch, Christian Müller, Achim G. Beule, and Jan-Peter Hildebrandt. 2019. "Sphingomyelin Depletion from Plasma Membranes of Human Airway Epithelial Cells Completely Abrogates the Deleterious Actions of S. aureus Alpha-Toxin" Toxins 11, no. 2: 126. https://doi.org/10.3390/toxins11020126