1. Peptide surfactants for cell-free production of functional G protein-coupled receptors.
- Author
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Wang X, Corin K, Baaske P, Wienken CJ, Jerabek-Willemsen M, Duhr S, Braun D, and Zhang S
- Subjects
- Amines chemistry, Cell-Free System, Circular Dichroism, Escherichia coli metabolism, Humans, Hydrogen-Ion Concentration, Ligands, Molecular Conformation, Olfactory Receptor Neurons metabolism, Protein Biosynthesis, Protein Structure, Secondary, Solubility, Peptides chemistry, Receptors, G-Protein-Coupled chemistry, Surface-Active Agents chemistry
- 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.
- Published
- 2011
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