title: Rehosting of bacterial chaperones for high-quality protein production.
abstract: Coproduction of DnaK/DnaJ in Escherichia coli enhances solubility but promotes proteolytic degradation of their substrates, minimizing the yield of unstable polypeptides. Higher eukaryotes have orthologs of DnaK/DnaJ but lack the linked bacterial proteolytic system. By coexpression of DnaK and DnaJ in insect cells with inherently misfolding-prone recombinant proteins, we demonstrate simultaneous improvement of soluble protein yield and quality and proteolytic stability. Thus, undesired side effects of bacterial folding modulators can be avoided by appropriate rehosting in heterologous cell expression systems.
authors.affiliation: Institute for Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Martínez-Alonso
authors.forename: Verónica
authors.initials: V
authors.lastname: Toledo-Rubio
authors.forename: Rob
authors.initials: R
authors.lastname: Noad
authors.forename: Ugutz
authors.initials: U
authors.lastname: Unzueta
authors.forename: Neus
authors.initials: N
authors.lastname: Ferrer-Miralles
authors.forename: Polly
authors.initials: P
authors.lastname: Roy
authors.forename: Antonio
authors.initials: A
authors.lastname: Villaverde
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D050956:HSP40 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology / Q000235:genetics*
chemical_list: C494635:DnaJ protein, E coli; D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; C063842:dnaK protein, E coli