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Journal Article

Semliki Forest Virus vectors for overexpression of 101 G Protein-coupled receptors in mammalian host cells

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Prual,  Cécile
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

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André,  Nicolas
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Reinhart,  Christoph
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Hassaine, G., Wagner, R., Kempf, J., Cherouati, N., Hassaine, N., Prual, C., et al. (2006). Semliki Forest Virus vectors for overexpression of 101 G Protein-coupled receptors in mammalian host cells. Protein Expression and Purification, 45(2), 343-351. doi:10.1016/j.pep.2005.06.007.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0024-D985-B
Abstract
Semliki Forest virus vectors were applied for the evaluation of 101 G protein-coupled receptors in three mammalian cell lines. Western blotting demonstrated that 95 of the 101 tested GPCRs showed positive signals. A large number of the GPCRs were expressed at high levels suggesting receptor yields in the range of 1 mg/L or higher, suitable for structural biology applications. Specific binding assays on a selected number of GPCRs were carried out to compare the correlation between total and functional protein expression. Ligands and additives supplemented to the cell culture medium were evaluated for expression enhancement. Selected GPCRs were also expressed from mutant SFV vectors providing enhanced protein expression and reduced host cell toxicity in attempts to further improve receptor yields.