The method utilized to purify the SARS-CoV-2 N protein can affect its molecular properties.
Gel-like structures
Intrinsically disordered protein (IDP)
Liquid-liquid phase separation (LLPS)
Journal
International journal of biological macromolecules
ISSN: 1879-0003
Titre abrégé: Int J Biol Macromol
Pays: Netherlands
ID NLM: 7909578
Informations de publication
Date de publication:
01 Oct 2021
01 Oct 2021
Historique:
received:
22
05
2021
revised:
02
08
2021
accepted:
03
08
2021
pubmed:
10
8
2021
medline:
30
9
2021
entrez:
9
8
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 34371045
pii: S0141-8130(21)01685-8
doi: 10.1016/j.ijbiomac.2021.08.026
pmc: PMC8343380
pii:
doi:
Substances chimiques
Coronavirus Nucleocapsid Proteins
0
Intrinsically Disordered Proteins
0
Nucleic Acids
0
Protein Aggregates
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
391-403Informations de copyright
Copyright © 2021. Published by Elsevier B.V.
Références
Cell Host Microbe. 2014 Oct 8;16(4):462-72
pubmed: 25299332
Nat Commun. 2020 Nov 27;11(1):6041
pubmed: 33247108
Molecules. 2020 Nov 02;25(21):
pubmed: 33147821
Biophys J. 2006 Jun 15;90(12):4651-61
pubmed: 16565040
J Struct Biol. 2011 Apr;174(1):11-22
pubmed: 21130884
Virology. 2007 May 10;361(2):304-15
pubmed: 17210170
J Med Virol. 2020 Jun;92(6):618-631
pubmed: 32108359
Biochem Biophys Res Commun. 2020 Jun 30;527(3):618-623
pubmed: 32416961
Biophys J. 2021 Jul 20;120(14):2771-2784
pubmed: 34214535
J Virol. 2019 Sep 12;93(19):
pubmed: 31315999
Protein Expr Purif. 2007 May;53(1):40-50
pubmed: 17251035
J Virol. 2006 Apr;80(7):3180-8
pubmed: 16537586
Nat Commun. 2021 Jan 21;12(1):502
pubmed: 33479198
J Infect Dis. 2020 Jun 29;222(2):206-213
pubmed: 32427334
Mol Cell. 2020 Dec 17;80(6):1078-1091.e6
pubmed: 33290746
J Biol Chem. 2009 Feb 20;284(8):5229-39
pubmed: 19106108
Biophys J. 2000 Mar;78(3):1606-19
pubmed: 10692345
Eur J Biochem. 2000 Jan;267(2):507-19
pubmed: 10632720
J Immunol Res. 2020 Sep 17;2020:2837670
pubmed: 32964056
Nature. 1970 Aug 15;227(5259):680-5
pubmed: 5432063
N Engl J Med. 2020 Feb 20;382(8):727-733
pubmed: 31978945
Curr Pharm Des. 2013;19(23):4191-213
pubmed: 23170892
Cell Res. 2020 Dec;30(12):1143-1145
pubmed: 32901111
Commun Biol. 2020 Dec 15;3(1):773
pubmed: 33319830
Viruses. 2014 Aug 07;6(8):2991-3018
pubmed: 25105276
Acta Pharmacol Sin. 2020 Sep;41(9):1141-1149
pubmed: 32747721
Nat Rev Microbiol. 2021 Mar;19(3):155-170
pubmed: 33116300
Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2017 Jul;9(4):
pubmed: 27813275
Acta Pharm Sin B. 2020 Jul;10(7):1228-1238
pubmed: 32363136
Biochem Biophys Res Commun. 2004 May 14;317(4):1030-6
pubmed: 15094372
Biochim Biophys Acta. 1997 Apr 4;1338(2):161-85
pubmed: 9128135
Future Virol. 2018 Jun;13(6):405-430
pubmed: 32201497
Science. 2009 Jun 26;324(5935):1729-32
pubmed: 19460965
Protein Sci. 2020 Sep;29(9):1890-1901
pubmed: 32654247
N Engl J Med. 2021 Mar 8;384(15):1466-1468
pubmed: 33684280
Adv Virus Res. 2006;66:193-292
pubmed: 16877062
J Virol Methods. 2005 Dec;130(1-2):45-50
pubmed: 16024098
Mol Cell. 2020 Dec 17;80(6):1092-1103.e4
pubmed: 33248025
J Phys Chem A. 2016 Nov 17;120(45):9008-9018
pubmed: 27809523
Mol Cell. 2015 Oct 15;60(2):208-19
pubmed: 26412307
Emerg Microbes Infect. 2020 Dec;9(1):313-319
pubmed: 32020836
Cell. 2020 Aug 6;182(3):685-712.e19
pubmed: 32645325
Nat Microbiol. 2020 Apr;5(4):562-569
pubmed: 32094589
Biochem Biophys Res Commun. 2003 Nov 28;311(4):870-6
pubmed: 14623261
Methods Enzymol. 1995;246:19-34
pubmed: 7538624
Adv Colloid Interface Sci. 2017 Jan;239:88-96
pubmed: 27393642
J Virol. 2007 Jan;81(2):548-57
pubmed: 17108024
J Virol. 2007 Apr;81(8):3913-21
pubmed: 17229691
Annu Rev Cell Dev Biol. 2014;30:39-58
pubmed: 25288112
Pathog Immun. 2020 Oct 19;5(1):342-363
pubmed: 33154981
Biochemistry. 2018 May 1;57(17):2415-2423
pubmed: 29323488
Cell Host Microbe. 2020 Apr 8;27(4):671-680.e2
pubmed: 32183941
Biochem J. 2004 Oct 1;383(Pt 1):13-8
pubmed: 15294014
Virus Res. 2004 Oct;105(2):121-5
pubmed: 15351485
J Med Virol. 2021 Apr;93(4):1923-1925
pubmed: 33386773
Proteins. 2006 Jan 1;62(1):24-45
pubmed: 16287116
Virology. 2019 Nov;537:198-207
pubmed: 31505321
Biochem Biophys Res Commun. 2021 Jan 29;538:54-62
pubmed: 33039147
Protein Expr Purif. 2005 Sep;43(1):1-9
pubmed: 16084395
Front Immunol. 2020 Oct 28;11:587615
pubmed: 33193414
EMBO J. 2020 Dec 15;39(24):e106478
pubmed: 33200826
FEBS J. 2008 Aug;275(16):4152-63
pubmed: 18631359
Arch Virol. 1992;125(1-4):141-60
pubmed: 1322650
FEBS Lett. 2005 Apr 25;579(11):2387-96
pubmed: 15848177
Biochim Biophys Acta. 2005 Aug 10;1751(2):119-39
pubmed: 16027053
Int J Biochem Cell Biol. 2006;38(8):1417-28
pubmed: 16546436
Science. 2003 May 30;300(5624):1399-404
pubmed: 12730501
J Immunol. 2008 Nov 1;181(9):6337-48
pubmed: 18941225
Virology. 1990 Nov;179(1):463-8
pubmed: 2171216
J Biomed Sci. 2006 Jan;13(1):59-72
pubmed: 16228284
Nat Commun. 2021 Mar 29;12(1):1936
pubmed: 33782395
Protein Expr Purif. 2005 May;41(1):207-34
pubmed: 15915565
J Mol Biol. 2021 Apr 16;433(8):166875
pubmed: 33556408
Virology. 2015 Apr;478:75-85
pubmed: 25726972