title: Peptide surfactants for cell-free production of functional G protein-coupled receptors.
abstract: Two major bottlenecks in elucidating the structure and function of membrane proteins are the difficulty of producing large quantities of functional receptors, and stabilizing them for a sufficient period of time. Selecting the right surfactant is thus crucial. Here we report using peptide surfactants in commercial Escherichia coli cell-free systems to rapidly produce milligram quantities of soluble G protein-coupled receptors (GPCRs). These include the human formyl peptide receptor, human trace amine-associated receptor, and two olfactory receptors. The GPCRs expressed in the presence of the peptide surfactants were soluble and had α-helical secondary structures, suggesting that they were properly folded. Microscale thermophoresis measurements showed that one olfactory receptor expressed using peptide surfactants bound its known ligand heptanal (molecular weight 114.18). These short and simple peptide surfactants may be able to facilitate the rapid production of GPCRs, or even other membrane proteins, for structure and function studies.
authors.affiliation: Center for Biomedical Engineering NE47-379, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139-4307, USA.
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Wang
authors.forename: Karolina
authors.initials: K
authors.lastname: Corin
authors.forename: Philipp
authors.initials: P
authors.lastname: Baaske
authors.forename: Christoph J
authors.initials: CJ
authors.lastname: Wienken
authors.forename: Moran
authors.initials: M
authors.lastname: Jerabek-Willemsen
authors.forename: Stefan
authors.initials: S
authors.lastname: Duhr
authors.forename: Dieter
authors.initials: D
authors.lastname: Braun
authors.forename: Shuguang
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000588:Amines / Q000737:chemistry; D002474:Cell-Free System; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008968:Molecular Conformation; D018034:Olfactory Receptor Neurons / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D000588:Amines; D008024:Ligands; D010455:Peptides; D043562:Receptors, G-Protein-Coupled; D013501:Surface-Active Agents