Interactions of the Lysosomotropic Detergent O-Methyl-Serine Dodecylamide Hydrochloride (MSDH) with Lipid Bilayer Membranes—Implications for Cell Toxicity
Abstract
:1. Introduction
2. Results
2.1. MSDH-induced Leakage of Small and Large Molecules is Affected by pH and Lipid Composition
2.2. MSDH Causes Permeabilization of Cellular Membranes and Cell Death
2.3. Uptake of MSDH by Endocytosis
3. Discussion
4. Materials and Methods
4.1. MSDH
4.2. Preparation of Liposomes
4.3. Fluorophore Leakage Experiments
4.4. FCS Measurements
4.5. Cells and Culture Conditions
4.6. Viability and Apoptosis Analysis
4.7. Immunocytochemistry
4.8. Cytosolic Extraction
4.9. Determination of β-N-acetyl-glucosaminidase (NAG) Activity
4.10. Western Blot
4.11. TEM
4.12. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ANTS | 8-Aminonapthalene-1,3,6 trisulfonic acid |
DPX | p-Xylene-bis-pyridinium bromide |
FCS | Fluorescence correlation spectroscopy |
LAMP2 | Lysosomal-associated membrane protein-2 |
LD | Lysosomotropic detergent |
LDH | Lactate dehydrogenase |
LLOMe | L-leucyl-L-leucine methyl ester |
LMP | Lysosomal membrane permeabilization |
MDC | Mono-dansylcadaverine |
MSDH | O-methyl-serine dodecylamine hydrochloride |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
NAG | β-N-acetyl-glucosaminidase |
TRITC-dextran 40 | Tetra-methyl rhodamine isothiocyanate–dextran with MW 40 kDa |
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Detergent | Liposomes pH 5 (milliseconds) | Liposomes pH 7 (milliseconds) | ||
---|---|---|---|---|
DOPC/DOPE | +Cholesterol | DOPC/DOPE | +Cholesterol | |
Triton X-100 | 0.30 ± 0.01 | 0.33 ± 0.01 | 0.28 ± 0.01 | 0.30 ± 0.01 |
MSDH:lipid 20:1 | 0.38 ± 0.07 | 0.48 ± 0.07 | 0.34 ± 0.04 | 0.51 ± 0.02* |
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Villamil Giraldo, A.-M.; Eriksson, I.; Wennmalm, S.; Fyrner, T.; Ederth, T.; Öllinger, K. Interactions of the Lysosomotropic Detergent O-Methyl-Serine Dodecylamide Hydrochloride (MSDH) with Lipid Bilayer Membranes—Implications for Cell Toxicity. Int. J. Mol. Sci. 2020, 21, 3136. https://doi.org/10.3390/ijms21093136
Villamil Giraldo A-M, Eriksson I, Wennmalm S, Fyrner T, Ederth T, Öllinger K. Interactions of the Lysosomotropic Detergent O-Methyl-Serine Dodecylamide Hydrochloride (MSDH) with Lipid Bilayer Membranes—Implications for Cell Toxicity. International Journal of Molecular Sciences. 2020; 21(9):3136. https://doi.org/10.3390/ijms21093136
Chicago/Turabian StyleVillamil Giraldo, Ana-Maria, Ida Eriksson, Stefan Wennmalm, Timmy Fyrner, Thomas Ederth, and Karin Öllinger. 2020. "Interactions of the Lysosomotropic Detergent O-Methyl-Serine Dodecylamide Hydrochloride (MSDH) with Lipid Bilayer Membranes—Implications for Cell Toxicity" International Journal of Molecular Sciences 21, no. 9: 3136. https://doi.org/10.3390/ijms21093136