2016 - Functional characterization of p7 viroporin from hepatitis C virus produced in a cell-free expression system
Port-a-Patch and Vesicle Prep Pro publication in Protein Expression and Purification (2016)
Soranzo T., Cortès S., Gilde F., Kreir M., Picart C., Lenormand J.-L.
Protein Expr Purif (2016) 118:83-91
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. Photoactivatable 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.