Membrane Activity and Viroporin Assembly for the SARS-CoV-2 E Protein Are Regulated by Cholesterol
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Formation of Giant Unilamellar Vesicles (GUVs)
2.3. Confocal Fluorescence Microscopy
2.4. Calcein Leakage Assay
2.5. Formation of Planar Bilayer Lipid Membranes (BLMs)
2.6. Patch-Clamp Measurements
2.7. Formation of Small Liposomes
2.8. Atomic Force Microscopy
3. Results
3.1. E Protein but Not Its H3 Peptide Deform GUVs with Charged Lipids and High Cholesterol Content
3.2. E Protein Induces Calcein Leakage from GUVs with Charged Lipids
3.3. Patch-Clamp Experiments Reveal Different Types of E Protein-Induced Membrane Conductance
3.4. Atomic Force Microscopy Demonstrates the Ability of the E Protein to Form Semi-Transmembrane or Transmembrane Pores Depending on the Cholesterol Content of a Membrane
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Volovik, M.V.; Denieva, Z.G.; Gifer, P.K.; Rakitina, M.A.; Batishchev, O.V. Membrane Activity and Viroporin Assembly for the SARS-CoV-2 E Protein Are Regulated by Cholesterol. Biomolecules 2024, 14, 1061. https://doi.org/10.3390/biom14091061
Volovik MV, Denieva ZG, Gifer PK, Rakitina MA, Batishchev OV. Membrane Activity and Viroporin Assembly for the SARS-CoV-2 E Protein Are Regulated by Cholesterol. Biomolecules. 2024; 14(9):1061. https://doi.org/10.3390/biom14091061
Chicago/Turabian StyleVolovik, Marta V., Zaret G. Denieva, Polina K. Gifer, Maria A. Rakitina, and Oleg V. Batishchev. 2024. "Membrane Activity and Viroporin Assembly for the SARS-CoV-2 E Protein Are Regulated by Cholesterol" Biomolecules 14, no. 9: 1061. https://doi.org/10.3390/biom14091061