Stabilization of full-length S-protein of SARS-Cov-2 coronavirus in SMA polymer for electron microscopy study
https://doi.org/10.55959/MSU0137-0952-16-78-3S-5
Abstract
A detergent-free protocol for purification of the coronavirus prefusion S-protein using styrenemaleic acid copolymer (SMA) was developed. Expression of the S-protein was carried out in HEK293T cells. Two solubilization methods were used to purify and prepare the S-protein for microscopy: in NP-40 detergent and as part of SMA. The resulting preparations were examined in an electron microscope, and the particles of purified S-proteins were classified. Analysis of two-dimensional projections of the particles showed that the use of lipodiscs for solubilization leads to lower mobility of the purified protein on the substrate, compared to the protein in the detergent, which may further contribute to obtaining higher resolutions when studying the structure of membrane proteins.
Keywords
electron microscopy,
structural biology,
single particle analysis,
SARS-CoV-2,
solubilization,
detergent,
SMA
Supporting agencies: The research was supported by MSU Development Program, project number 23A-Sh04-01.
About the Authors
N. Yu. Mamaeva
Moscow State University
Russian Federation
Russian Federation
Faculty of Biology
1–73 Leninskie Gory, 119234, Moscow
N. I. Derkacheva
A.I. Evdokimov Moscow State Medical and Dental University
Russian Federation
Russian Federation
Department of Biochemistry
4 Dolgorukovskaya st., 127006, Moscow
D. A. Gasanova
Lomonosov Moscow State University
Russian Federation
Russian Federation
3Department of Chemical Enzymology, Faculty of Chemistry
1–11 Leninskie Gory, 119234, Moscow
O. S. Sokolova
Lomonosov Moscow State University
Russian Federation
Russian Federation
Faculty of Biology
1–73 Leninskie Gory, 119234, Moscow
G. S. Glukhov
Shenzhen MSU-BIT University, 1 International University Park Road
China
China
Faculty of Biology
Dayun New Town, Longgang District, Shenzhen, 518172, Guangdong Province
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Review
For citations:
Mamaeva N.Yu., Derkacheva N.I., Gasanova D.A., Sokolova O.S., Glukhov G.S. Stabilization of full-length S-protein of SARS-Cov-2 coronavirus in SMA polymer for electron microscopy study. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2023;78(3S):27-32. (In Russ.) https://doi.org/10.55959/MSU0137-0952-16-78-3S-5
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