Enhanced expression and purification of membrane proteins by SUMO fusion in Escherichia coli.
Severe acute respiratory syndrome coronavirus (SARS-CoV) membrane protein and 5-lipoxygenase-activating protein (FLAP) are among a large number of membrane proteins that are poorly expressed when traditional expression systems and methods are employed. Therefore to efficiently express difficult membrane proteins, molecular biologists will have to develop novel or innovative expression systems. To this end, we have expressed the SARS-CoV M and FLAP proteins in Escherichia coli by utilizing a novel gene fusion expression system that takes advantage of the natural chaperoning properties of the SUMO (small ubiquitin-related modifier) tag. These chaperoning properties facilitate proper protein folding, which enhances the solubility and biological activity of the purified protein. In addition to these advantages, we found that SUMO Protease 1, can cleave the SUMO fusion high specificity to generate native protein. Herein, we demonstrate that the expression of FLAP and SARS-CoV membrane proteins are greatly enhanced by SUMO fusions in E. coli.
Zuo
Xun
X
LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USA.
Li
Shuisen
S
Hall
John
J
Mattern
Michael R
MR
Tran
Hiep
H
Shoo
Joshua
J
Tan
Robin
R
Weiss
Susan R
SR
Butt
Tauseef R
TR
D058927:5-Lipoxygenase-Activating Proteins; D015153:Blotting, Western; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D000086862:Coronavirus M Proteins; D017931:DNA Primers; D030562:Databases, Protein; D004926:Escherichia coli / Q000737:chemistry*; D005822:Genetic Vectors / Q000235:genetics; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D017510:Protein Folding; D025842:SUMO-1 Protein / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000302:isolation & purification* / Q000378:metabolism
D058927:5-Lipoxygenase-Activating Proteins; D002352:Carrier Proteins; D000086862:Coronavirus M Proteins; D017931:DNA Primers; D008565:Membrane Proteins; D025842:SUMO-1 Protein; D014763:Viral Matrix Proteins