Expression of proteins in insect cells using recombinant baculoviruses has gained wide use in the G protein-coupled receptor (GPCR) community. This expression system produces high yields of functional receptor, is able to perform post-translational modifications, and is readily adaptable to large-scale culture. Here, we describe the generic methods for expressing a GPCR using baculovirus-infected insect cells, including the maintenance of insect cell culture. Data are presented for polyhedrin promoter-driven expression of a C-terminal 6 x histidine-tagged mammalian M(2) muscarinic receptor in Sf9 cells. Results demonstrate that expressed receptor could be detected and quantified using radiolabeled ligand binding, that expression was maximal at approximately 72 h post-infection, and that expression levels could be altered by addition of various ligands to cultures of infected insect cells.
CITATION STYLE
Aloia, A. L., Glatz, R. V., McMurchie, E. J., & Leifert, W. R. (2009). GPCR expression using baculovirus-infected Sf9 cells. Methods in Molecular Biology (Clifton, N.J.), 552, 115–129. https://doi.org/10.1007/978-1-60327-317-6_8
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