Enhanced Expression and Purification of Membrane Proteins by SUMO Fusion in Escherichia coli |
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Authors: | Xun Zuo Shuisen Li John Hall Michael R Mattern Hiep Tran Joshua Shoo Robin Tan Susan R Weiss Tauseef R Butt |
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Institution: | (1) LifeSensors, Inc., 271 Great Valley Parkway, Malvern, PA 19355, USA;(2) School of BioMedical Engineering, Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104, USA;(3) Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore, 119260;(4) Department of Microbiology, University of Pennsylvania Medical School, Philadelphia, PA 19104, USA |
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Abstract: | 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. |
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Keywords: | 5-lipoxygenase activating protein (FLAP) membrane protein expression Nickel affinity purification SARS-CoV membrane protein SUMO fusion |
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