Abstract
Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 μM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.
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ACKNOWLEDGMENTS
The authors are grateful to E.S. Bunin for assistance in performing Western blot analysis and A.S. Saburov for help in isolating proteins. The experiments were performed using the equipment of the Core Facility of the Institute of Gene Biology, Russian Academy of Sciences.
Funding
The study was supported by the Russian Science Foundation (project no. 21-14-00130).
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Translated by M. Batrukova
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Khramtsov, Y.V., Ulasov, A.V., Lupanova, T.N. et al. Quantitative Description of the N-Protein of the SARS-CoV-2 Virus Degradation in Cells Stably Expressing It under the Influence of New Modular Nanotransporters. Dokl Biochem Biophys 513 (Suppl 1), S63–S66 (2023). https://doi.org/10.1134/S1607672923700709
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DOI: https://doi.org/10.1134/S1607672923700709