Abstract
On the basis of literature data, an antibody-like molecule, monobody, was selected that is capable of interacting with the nucleocapsid protein (N protein) of the SARS-CoV-2 virus with a high affinity (dissociation constant 6.7 nM). We have previously developed modular nanotransporters (MNTs) to deliver various molecules to a selected compartment of target cells. In this work, a monobody to the N protein of the SARS-CoV-2 virus was inserted in the MNT using genetic engineering methods. In this MNT, a site for the cleavage of the monobody from the MNT in endosomes was also inserted. It was shown by thermophoresis that the cleavage of this monobody from the MNT by the endosomal protease cathepsin B leads to a 12-fold increase in the affinity of the monobody for the N protein. Cellular thermal shift assay showed the ability of the obtained MNT to interact with the N protein in A431 cells transfected with the SARS-CoV-2 N protein fused to the mRuby3 fluorescent protein.
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ACKNOWLEDGMENTS
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 nos. 22-24-00035 and 21-14-00130): all experiments related to the monobody cleavage by cathepsin B were performed under the project no. 22-24-00035, and all other experiments were performed under the project no. 21-14-00130.
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The authors declare that they have no conflicts of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.
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Translated by M. Batrukova
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Khramtsov, Y.V., Ulasov, A.V., Lupanova, T.N. et al. Modular Nanotransporters Capable of Binding to SARS-CoV-2 Virus Nucleocapsid Protein in Target Cells. Dokl Biochem Biophys 510, 87–90 (2023). https://doi.org/10.1134/S1607672922600233
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DOI: https://doi.org/10.1134/S1607672922600233