title: A novel expression system for production of soluble prion proteins in E. coli.
abstract: Expression of eukaryotic proteins in Escherichia coli is challenging, especially when they contain disulfide bonds. Since the discovery of the prion protein (PrP) and its role in transmissible spongiform encephalopathies, the need to obtain large quantities of the recombinant protein for research purposes has been essential. Currently, production of recombinant PrP is achieved by refolding protocols. Here, we show that the co-expression of two different PrP with the human Quiescin Sulfhydryl OXidase (QSOX), a human chaperone with thiol/disulfide oxidase activity, in the cytoplasm of E. coli produces soluble recombinant PrP. The structural integrity of the soluble PrP has been confirmed by nuclear magnetic resonance spectroscopy, demonstrating that properly folded PrP can be easily expressed in bacteria. Furthermore, the soluble recombinant PrP produced with this method can be used for functional and structural studies.
authors.affiliation: VIB, Department of Structural Biology, Brussels, Belgium.
authors.forename: Romany N N
authors.initials: RN
authors.lastname: Abskharon
authors.forename: Stephanie
authors.initials: S
authors.lastname: Ramboarina
authors.forename: Hassan
authors.initials: H
authors.lastname: El Hassan
authors.forename: Wael
authors.initials: W
authors.lastname: Gad
authors.forename: Marcin I
authors.initials: MI
authors.lastname: Apostol
authors.forename: Gabriele
authors.initials: G
authors.lastname: Giachin
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Legname
authors.forename: Jan
authors.initials: J
authors.lastname: Steyaert
authors.forename: Joris
authors.initials: J
authors.lastname: Messens
authors.forename: Sameh H
authors.initials: SH
authors.lastname: Soror
authors.forename: Alexandre
authors.initials: A
authors.lastname: Wohlkonig
mesh_terms: D001709:Biotechnology / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000235:genetics / Q000378:metabolism; D011328:Prions / Q000096:biosynthesis* / Q000235:genetics; D011490:Protein Disulfide Reductase (Glutathione) / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011328:Prions; D011994:Recombinant Proteins; D050862:Oxidoreductases Acting on Sulfur Group Donors; C117183:QSOX1 protein, human; D011490:Protein Disulfide Reductase (Glutathione)