التفاصيل البيبلوغرافية
| العنوان: |
Functional expression, purification, characterization, and membrane reconstitution of non-structural protein 2 from hepatitis C virus. |
| المؤلفون: |
Fogeron, Marie-Laure1, Paul, David2,3, Jirasko, Vlastimil2,3, Montserret, Roland1, Lacabanne, Denis1, Molle, Jennifer1, Badillo, Aurélie1,4, Boukadida, Célia5, Georgeault, Sonia6, Roingeard, Philippe6,7, Martin, Annette5, Bartenschlager, Ralf2,3, Penin, François1 f.penin@ibcp.fr, Böckmann, Anja1 a.bockmann@ibcp.fr |
| المصدر: |
Protein Expression & Purification. Dec2015, Vol. 116, p1-6. 6p. |
| مصطلحات موضوعية: |
*PROTEIN expression, *CHEMICAL purification, *MEMBRANE proteins, *PROTEIN structure, *HEPATITIS C virus, *CYSTEINE proteinases |
| مستخلص: |
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. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
Academic Search Index |
