A scalable, GFP-based pipeline for membrane protein overexpression screening and purification.
Drew D., Slotboom D-J., Friso G., Reda T., Genevaux P., Rapp M., Meindl-Beinker NM., Lambert W., Lerch M., Daley DO., Van Wijk K-J., Hirst J., Kunji E., De Gier J-W.
We describe a generic, GFP-based pipeline for membrane protein overexpression and purification in Escherichia coli. We exemplify the use of the pipeline by the identification and characterization of E. coli YedZ, a new, membrane-integral flavocytochrome. The approach is scalable and suitable for high-throughput applications. The GFP-based pipeline will facilitate the characterization of the E. coli membrane proteome and serves as an important reference for the characterization of other membrane proteomes.