A scalable, GFP-based pipeline for membrane protein overexpression screening and purification.
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.
Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
David
D
Drew
Dirk-Jan
DJ
Slotboom
Giulia
G
Friso
Torsten
T
Reda
Pierre
P
Genevaux
Mikaela
M
Rapp
Nadja M
NM
Meindl-Beinker
Wietske
W
Lambert
Mirjam
M
Lerch
Daniel O
DO
Daley
Klaas-Jan
KJ
Van Wijk
Judy
J
Hirst
Edmund
E
Kunji
Jan-Willem
JW
De Gier
D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics; D049408:Luminescent Agents / Q000032:analysis*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D029968:Escherichia coli Proteins; D049408:Luminescent Agents; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins