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Functional expression, purification, characterization, and membrane reconstitution of non-structural protein 2 from hepatitis C virus

Non-structural protein 2 (NS2) of the hepatitis C virus (HCV) is an integral membrane protein that contains a cysteine protease and that plays a central organizing role in assembly of infectious progeny virions. While the crystal structure of the protease domain has been solved, the NS2 full-length...

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Bibliographic Details
Main Authors: Fogeron, Marie-Laure (Author) , Paul, David (Author) , Jirasko, Vlastimil (Author) , Bartenschlager, Ralf (Author)
Format: Article (Journal)
Language:English
Published: 29 August 2015
In: Protein expression and purification
Year: 2015, Volume: 116, Pages: 1-6
ISSN:1096-0279
DOI:10.1016/j.pep.2015.08.027
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.pep.2015.08.027
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S1046592815300474
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Author Notes:Marie-Laure Fogeron, David Paul, Vlastimil Jirasko, Roland Montserret, Denis Lacabanne, Jennifer Molle, Aurélie Badillo, Célia Boukadida, Sonia Georgeault, Philippe Roingeard, Annette Martin, Ralf Bartenschlager, François Penin, Anja Böckmann
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Summary:Non-structural protein 2 (NS2) of the hepatitis C virus (HCV) is an integral membrane protein that contains a cysteine protease and that plays a central organizing role in assembly of infectious progeny virions. While the crystal structure of the protease domain has been solved, the NS2 full-length form remains biochemically and structurally uncharacterized because recombinant NS2 could not be prepared in sufficient quantities from cell-based systems. We show here that functional NS2 in the context of the NS2-NS3pro precursor protein, ensuring NS2-NS3 cleavage, can be efficiently expressed by using a wheat germ cell-free expression system. In this same system, we subsequently successfully produce and purify milligram amounts of a detergent-solubilized form of full-length NS2 exhibiting the expected secondary structure content. Furthermore, immuno-electron microscopy analyses of reconstituted proteoliposomes demonstrate NS2 association with model membranes.
Item Description:Gesehen am 03.08.2021
Physical Description:Online Resource
ISSN:1096-0279
DOI:10.1016/j.pep.2015.08.027

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