Cholesterol Increases Lipid Binding Rate and Changes Binding Behavior of Bacillus thuringiensis Cytolytic Protein
Abstract
:1. Introduction
2. Results
2.1. Changes in Lipid Bilayer Properties Upon Increasing of Cholesterol
2.2. Increasing of Protein Adsorption Rate and Lipid Binding Behavior as Function of Cholesterol Content
2.3. Nanostructure of the Cyt2Aa2/Lipid Bilayer
3. Discussion
3.1. The Role of Cholesterol on Lipid Bilayer
3.2. Protein Binding as a Function of Cholesterol Content in Lipid Bilayer
4. Materials and Methods
4.1. Reagents and Buffer
4.2. Protein Preparation
4.3. Liposome Preparation and Characterization
4.4. Quartz Crystal Microbalance with Dissipation (QCM-D) Measurement
4.5. Atomic Force Microscope (AFM) Imaging
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
Bt | Bacillus thuringiensis |
Cyt protein | Cytolytic protein |
Cyt2Aa2 protein | Cytolytic protein from Bacillus thuringiensis subsp. darmstadiensis |
QCM-D | Quartz crystal microbalance with dissipation |
AFM | Atomic force microscope |
POPC | 1-palmitoyl,2-oleoyl-snglycero-3-phosphocholine |
Chol | cholesterol |
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POPC:Chol Ratio (by Weight) | ∆F5 (Hz) | ∆D5 (×10−6) |
---|---|---|
5:0 | −26.1 ± 1.2 | 1.6 ± 0.9 |
5:0.2 | −29.6 ± 2.6 | 1.6 ± 0.6 |
5:1 | −30.3 ± 0.5 | 1.9 ± 0.1 |
5:2 | −37.1 ± 1.0 | 3.6 ± 0.5 |
5:3 | −31.3 ± 0.6 | 2.2 ± 0.1 |
POPC:Chol Ratio (by Weight) | ∆F5 (Hz) | ∆D5 (×10−6) | Γ (min) |
---|---|---|---|
5:0 | −29 ± 2 | 2.2 ± 0.7 | 6.9 |
5:0.2 | −33 ± 2 | 2.2 ± 0.3 | 6.3 |
5:1 | −30 ± 1 | 2.7 ± 0.3 | 3.2 |
5:2 | −32 ± 1 | 2.5 ± 0.9 | 2.1 |
5:3 | −207 ± 4 | 43.7 ± 1.1 | N/A |
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Tharad, S.; Üzülmez, Ö.; Promdonkoy, B.; Toca-Herrera, J.L. Cholesterol Increases Lipid Binding Rate and Changes Binding Behavior of Bacillus thuringiensis Cytolytic Protein. Int. J. Mol. Sci. 2018, 19, 3819. https://doi.org/10.3390/ijms19123819
Tharad S, Üzülmez Ö, Promdonkoy B, Toca-Herrera JL. Cholesterol Increases Lipid Binding Rate and Changes Binding Behavior of Bacillus thuringiensis Cytolytic Protein. International Journal of Molecular Sciences. 2018; 19(12):3819. https://doi.org/10.3390/ijms19123819
Chicago/Turabian StyleTharad, Sudarat, Öykü Üzülmez, Boonhiang Promdonkoy, and José L. Toca-Herrera. 2018. "Cholesterol Increases Lipid Binding Rate and Changes Binding Behavior of Bacillus thuringiensis Cytolytic Protein" International Journal of Molecular Sciences 19, no. 12: 3819. https://doi.org/10.3390/ijms19123819