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.
MeSH Terms
Escherichia coli/genetics
Escherichia coli Proteins/biosynthesis,genetics,isolation & purification
Green Fluorescent Proteins/analysis,genetics
Lactococcus lactis/genetics
Luminescent Agents/analysis
Membrane Proteins/biosynthesis,genetics,isolation & purification
Recombinant Fusion Proteins/biosynthesis,isolation & purification
Chemicals
Escherichia coli Proteins
Luminescent Agents
Membrane Proteins
Recombinant Fusion Proteins
Green Fluorescent Proteins
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Drew David
Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
Slotboom Dirk-Jan
Friso Giulia
Reda Torsten
Genevaux Pierre
Rapp Mikaela
Meindl-Beinker Nadja M
Lambert Wietske
Lerch Mirjam
Daley Daniel O
Van Wijk Klaas-Jan
Hirst Judy
Kunji Edmund
De Gier Jan-Willem
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