title: Strategies for the expression of SUMO-modified target proteins in Escherichia coli.
abstract: We previously described the establishment of a binary vector system that allows co-expression of SUMO conjugation enzymes and a target protein of interest, leading to efficient SUMO modification and the production of a large amount of recombinant SUMO-modified proteins in Escherichia coli. The advantages of this E. coli expression/modification approach include scalability of experiments, low cost, fast growth, and a lack of proteases that cleave the isopeptide linkage between SUMO and the target protein. Thus, this E. coli method provides a useful alternative to authentic SUMO modification assays, such as in vitro SUMO conjugation and in vivo SUMO modification using baculovirus or mammalian cell culture, that are usually complicated, time-consuming and expensive.
authors.affiliation: Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan.
authors.forename: Hisato
authors.initials: H
authors.lastname: Saitoh
authors.forename: Junsuke
authors.initials: J
authors.lastname: Uwada
authors.forename: Kawasaki
authors.initials: K
authors.lastname: Azusa
mesh_terms: D000465:Algorithms; D019411:Clinical Laboratory Techniques; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D008828:Microbiological Techniques; D008954:Models, Biological; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial / Q000502:physiology
chemical_list: D011506:Proteins; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins