title: Cell-free expression of G-protein-coupled receptors.
abstract: Cell-free expression has emerged as a new standard for the production of membrane proteins. The reduction of expression complexity in cell-free systems eliminates central bottlenecks and allows the reliable and efficient synthesis of many different types of membrane proteins. Furthermore, the open accessibility of cell-free reactions enables the co-translational solubilization of cell-free expressed membrane proteins in a large variety of supplied additives. Hydrophobic environments can therefore be adjusted according to the requirements of individual membrane protein targets. We present different approaches for the preparative scale cell-free production of G-protein-coupled receptors using the extracts of Escherichia coli cells. We exemplify expression conditions implementing detergents, nanodiscs, or liposomes. The generated protein samples could be directly used for further functional characterization.
authors.affiliation: Centre for Biomolecular Magnetic Resonance, Institute of Biophysical Chemistry, Goethe-University Frankfurt/Main, Max-von-Laue-Str. 9, Frankfurt/Main, D-60438, Germany.
authors.forename: Erika
authors.initials: E
authors.lastname: Orbán
authors.forename: Davide
authors.initials: D
authors.lastname: Proverbio
authors.forename: Stefan
authors.initials: S
authors.lastname: Haberstock
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
authors.forename: Frank
authors.initials: F
authors.lastname: Bernhard
mesh_terms: D002457:Cell Extracts / Q000737:chemistry; D002474:Cell-Free System / Q000378:metabolism*; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry; D049329:Nanostructures / Q000737:chemistry; D011485:Protein Binding; D017510:Protein Folding; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis* / Q000302:isolation & purification*; D012995:Solubility
chemical_list: D002457:Cell Extracts; D003902:Detergents; D008081:Liposomes; D043562:Receptors, G-Protein-Coupled