Syracuse

Functional characterization of p7 viroporin from hepatitis C virus produced in a cell-free expression system

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Soranzo, T. | Cortes, S. | Gilde, Flora | Kreir, M. | Picart, C. | Lenormand, J. L.

Edité par CCSD ; Elsevier -

International audience. Using a cell-free expression system we produced the p7 viroporin embedded into a lipid bilayer in a single-step manner. The protein quality was assessed using different methods. We examined the channel forming activity of p7 and verified its inhibition by 5-(N,N-Hexamethylene) amiloride (HMA). Fourier transformed infrared spectroscopy (FTIR) experiments further showed that when p7 was inserted into synthetic liposomes, the protein displayed a native-like conformation similar to p7 obtained from other sources. Photoactivable amino acid analogs used for p7 protein synthesis enabled oligomerization state analysis in liposomes by cross-linking. Therefore, these findings emphasize the quality of the cell-free produced p7 proteoliposomes which can benefit the field of the hepatitis C virus (HCV) protein production and characterization and also provide tools for the development of new inhibitors to reinforce our therapeutic arsenal against HCV. (C) 2015 Elsevier Inc. All rights reserved.

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