Chapter title |
Wheat Germ Cell-Free Overexpression for the Production of Membrane Proteins
|
---|---|
Chapter number | 5 |
Book title |
Membrane Protein Structure and Function Characterization
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7151-0_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7149-7, 978-1-4939-7151-0
|
Authors |
Marie-Laure Fogeron, Aurélie Badillo, François Penin, Anja Böckmann |
Abstract |
Due to their hydrophobic nature, membrane proteins are notoriously difficult to express in classical cell-based protein expression systems. Often toxic, they also undergo degradation in cells or aggregate in inclusion bodies, making delicate issues further solubilization and renaturation. These are major bottlenecks in their structural and functional analysis. The wheat germ cell-free (WGE-CF) system offers an effective alternative not only to classical cell-based protein expression systems but also to other cell-free systems for the expression of membrane proteins. The WGE-CF indeed allows the production of milligram amounts of membrane proteins in a detergent-solubilized, homogenous, and active form. Here, we describe the method to produce a viral integral membrane protein, which is the non-structural protein 2 (NS2) of hepatitis C virus, in view of structural studies by solid-state NMR in a native-like lipid environment. |
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