A scalable, GFP-based pipeline for membrane protein overexpression screening and purification.

TitleA scalable, GFP-based pipeline for membrane protein overexpression screening and purification.
Publication TypeJournal Article
Year of Publication2005
AuthorsDrew, 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, ERS, de Gier, J-W
JournalProtein Sci
Volume14
Issue8
Pagination2011-7
Date Published2005 Aug
ISSN0961-8368
KeywordsEscherichia coli, Escherichia coli Proteins, Green Fluorescent Proteins, Lactococcus lactis, Luminescent Agents, Membrane Proteins, Recombinant Fusion Proteins
Abstract

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.

DOI10.1110/ps.051466205
Alternate JournalProtein Sci.
Citation Key10.1110/ps.051466205
PubMed ID15987891
PubMed Central IDPMC2279312