RT Journal Article
SR Electronic
T1 The method utilized to purify the SARS-CoV-2 N protein can affect its molecular properties
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.05.03.442392
DO 10.1101/2021.05.03.442392
A1 Aneta Tarczewska
A1 Marta Kolonko-Adamska
A1 Mirosław Zarębski
A1 Jurek Dobrucki
A1 Andrzej Ożyhar
A1 Beata Greb-Markiewicz
YR 2021
UL http://biorxiv.org/content/early/2021/05/04/2021.05.03.442392.abstract
AB One of the main structural proteins of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the nucleocapsid protein (N). The basic function of this protein is to bind genomic RNA and to form a protective nucleocapsid in the mature virion. The intrinsic ability of the N protein to interact with nucleic acids makes its purification very challenging. Therefore, typically employed purification methods appear to be insufficient for removing nucleic acid contamination. In this study, we present a novel purification protocol that enables the N protein to be prepared without any bound nucleic acids. We also performed comparative structural analysis of the N protein contaminated with nucleic acids and free of contamination and showed significant differences in the structural and phase separation properties of the protein. These results indicate that nucleic-acid contamination may severely affect molecular properties of the purified N protein. In addition, the notable ability of the N protein to form condensates whose morphology and behaviour suggest more ordered forms resembling gel-like or solid structures is described.