Expression of membrane proteins in Drosophila Melanogaster S2 cells: Production and analysis of a EGFP-fused G protein-coupled receptor as a model.
In the process of selecting an appropriate host for the heterologous expression of functional eukaryotic membrane proteins, Drosophila S2 cells, although not yet fully explored, appear as a valuable alternative to mammalian cell lines or other virus-infected insect cell systems. This nonlytic, plasmid-based system actually combines several major physiological and bioprocess advantages that make it a highly potential and scalable cellular tool for the production of membrane proteins in a variety of applications, including functional characterization, pharmacological profiling, molecular simulations, structural analyses, or generation of vaccines. We present here a series of protocols and hints that would serve the successful expression of membrane proteins in S2 cells, using an enhanced green fluorescent protein (EGFP)/G protein-coupled receptor (EGFP-GPCR) as a model.
Brillet
Karl
K
Dpt Récepteurs et des Protéines Membranaires, Illkirch, France.
Pereira
Carlos A
CA
Wagner
Renaud
R
D000818:Animals; D019149:Bioreactors; D015153:Blotting, Western; D002460:Cell Line; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism; D017450:Receptors, Opioid, mu / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D043562:Receptors, G-Protein-Coupled; D017450:Receptors, Opioid, mu; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins