Enhanced Expression and Purification of Membrane Proteins by SUMO Fusion in Escherichia coli
| Autor: | Hiep T. West Chester Tran, Michael R. Mattern, Robin Tan, Tauseef R. Butt, Joshua Shoo, Xun Zuo, Susan R. Weiss, John Hall, Shuisen Li |
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| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Vesicle-associated membrane protein 8
Protein Folding Coronavirus M Proteins medicine.medical_treatment 5-Lipoxygenase-Activating Proteins Blotting Western Genetic Vectors SUMO-1 Protein medicine.disease_cause Biochemistry Article Viral Matrix Proteins Structural Biology Cleave Protein A/G Genetics medicine Escherichia coli SARS-CoV membrane protein Databases Protein DNA Primers Protease Nickel affinity purification biology 5-lipoxygenase activating protein (FLAP) Membrane Proteins General Medicine membrane protein expression Blot Membrane protein biology.protein Protein folding SUMO fusion Carrier Proteins |
| Zdroj: | Journal of Structural and Functional Genomics |
| ISSN: | 1570-0267 1345-711X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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