title: Functional transplantation of the sumoylation machinery into Escherichia coli.
abstract: Modification by SUMO proteins appears to be very common in eukaryotic cells. Many proteins have been reported to be sumoylated, at least under certain circumstances, in vivo, and new examples get published every month. On the other hand, sumoylation is, in essence, a way to construct branched proteins or protein fusions. Obtention of pure sumoylated proteins from eukaryotic cells is not easy because of the dynamic nature of this modification and the large number of sumoylated proteins in vivo. Production of sumoylated proteins in vitro requires the previous purification of most of the components of the pathway, and has the typical limitations of such systems. In this paper, we describe a method to quantitatively produce sumoylated proteins in vivo in Escherichia coli as a way to obtain large quantities of specifically sumoylated target proteins with a high degree of purity, to generate fusion proteins not limited to N- or C-end additions, and to polymerize proteins by covalent linkage.
authors.affiliation: Department of Microbial Biotechnology, Centro Nacional de Biotecnología CSIC, Campus de Cantoblanco, 28049 Madrid, Spain. mmencia@cnb.uam.es
authors.forename: Mario
authors.initials: M
authors.lastname: Mencía
authors.forename: Víctor
authors.initials: V
authors.lastname: de Lorenzo
mesh_terms: D001671:Biochemistry / Q000379:methods*; D015153:Blotting, Western; D004247:DNA / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011506:Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry
chemical_list: D010455:Peptides; D011506:Proteins; D004247:DNA; D044763:Ubiquitin-Conjugating Enzymes; D005982:Glutathione Transferase