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© Michel Huerre
Coupe histologique de foie de patient atteint d'une hépatite C chronique active avec infiltrat (opacité) folliculaire (lymphocytes) (Grossissement X 400). L'hépatite C chronique peut être à l'origine de cirrhose et cancer du foie
Publication : Protein expression and purification

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

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Protein expression and purification - 29 Aug 2015

Fogeron ML, Paul D, Jirasko V, Montserret R, Lacabanne D, Molle J, Badillo A, Boukadida C, Georgeault S, Roingeard P, Martin A, Bartenschlager R, Penin F, Böckmann A

Link to Pubmed [PMID] – 26325423

Protein Expr. Purif. 2015 Dec;116:1-6

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.

https://www.ncbi.nlm.nih.gov/pubmed/26325423