Membrane Permeabilization by Bordetella Adenylate Cyclase Toxin Involves Pores of Tunable Size
Abstract
:1. Introduction
2. Materials and Methods
2.1. Antibodies and Reagents
2.2. ACT Purification
2.3. Cell Culture
2.4. Permeabilization Measured as Influx of Fluorescent Solutes to the Interior of Giant Unilamellar Vesicles (GUVs)
2.5. Atomic Force Microscopy (AFM): Sample Preparation, Measurements and Topographic Analysis
2.6. Preparation of Large Unilamellar Vesicles (LUVs)
2.7. Blue-Native (BN-PAGE) Electrophoresis and Western Blotting
2.8. Statistical Analysis
3. Results
3.1. Permeabilization of Giant Unilamellar Vesicles by ACT
3.2. ACT Forms Growing-Size Pores, Which Depend on Toxin Concentration and Incubation Time
3.3. Analysis by BN-PAGE and AFM of ACT Assemblies in Phosphatidylcholine Membranes
3.4. Permeabilization of Target Macrophages by ACT Directly Correlates with the Toxin Assembly into Oligomers of Variable Stoichiometry
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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González-Bullón, D.; B. Uribe, K.; Largo, E.; Guembelzu, G.; García-Arribas, A.B.; Martín, C.; Ostolaza, H. Membrane Permeabilization by Bordetella Adenylate Cyclase Toxin Involves Pores of Tunable Size. Biomolecules 2019, 9, 183. https://doi.org/10.3390/biom9050183
González-Bullón D, B. Uribe K, Largo E, Guembelzu G, García-Arribas AB, Martín C, Ostolaza H. Membrane Permeabilization by Bordetella Adenylate Cyclase Toxin Involves Pores of Tunable Size. Biomolecules. 2019; 9(5):183. https://doi.org/10.3390/biom9050183
Chicago/Turabian StyleGonzález-Bullón, David, Kepa B. Uribe, Eneko Largo, Garazi Guembelzu, Aritz B. García-Arribas, César Martín, and Helena Ostolaza. 2019. "Membrane Permeabilization by Bordetella Adenylate Cyclase Toxin Involves Pores of Tunable Size" Biomolecules 9, no. 5: 183. https://doi.org/10.3390/biom9050183