Nanodisc production in living cells facilitates structural studies of membrane proteins

Production of samples in high quality and quantity is the main bottleneck in structural studies of complex membrane proteins (MPs) from higher organisms. In addition, the majority of solved MP structures have been solved in the presence of detergents, which often hamper the activity and stability of MPs compared to a native lipid environment. The aim of this project is to develop a novel method independent of detergents, while still allowing for production of active MPs for structural studies. The approach is based on the production of nanodiscs (NDs) in living cells, and subsequent expression of the MPs directly into the NDs. NDs provide a lipid bilayer for the MPs and have been shown to maintain activity and stability to a higher degree than detergents. If successful, combining this technology with the high yields from expression in living cells will result in a superior production method and will greatly increase the accessibility of MP samples for structural studies in a native lipid environment.