Coupling neutron reflectivity with cell-free protein synthesis to probe membrane protein structure in supported bilayers.

ABSTRACT: The structure of the p7 viroporin, an oligomeric membrane protein ion channel involved in the assembly and release of the hepatitis C virus, was determined from proteins expressed and inserted directly into supported model lipid membranes using cell-free protein expression. Cell-free protein expression allowed (i ) high protein concentration in the membrane, (ii ) control of the protein's isotopic constitution, and (iii ) control over the lipid environment available to the protein. Here, we used cell-free protein synthesis to directly incorporate the hepatitis C virus (HCV) p7 protein into supported lipid bilayers formed from physiologically relevant lipids (POPC or asolectin) for both direct structural measurements using neutron reflectivity (NR) and conductance measurements using electri

SUBMITTER: Soranzo T 

PROVIDER: S-EPMC5469739 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature