title: The quiescin sulfhydryl oxidase (hQSOX1b) tunes the expression of resistin-like molecule alpha (RELM-α or mFIZZ1) in a wheat germ cell-free extract.
abstract: BACKGROUND
  Although disulfide bond formation in proteins is one of the most common types of post-translational modifications, the production of recombinant disulfide-rich proteins remains a challenge. The most popular host for recombinant protein production is Escherichia coli, but disulfide-rich proteins are here often misfolded, degraded, or found in inclusion bodies.
  
  
  RESULTS
  We optimize an in vitro wheat germ translation system for the expression of an immunological important eukaryotic protein that has to form five disulfide bonds, resistin-like alpha (mFIZZ1). Expression in combination with human quiescin sulfhydryl oxidase (hQSOX1b), the disulfide bond-forming enzyme of the endoplasmic reticulum, results in soluble, intramolecular disulfide bonded, monomeric, and biological active protein. The mFIZZ1 protein clearly suppresses the production of the cytokines IL-5 and IL-13 in mouse splenocytes cultured under Th2 permissive conditions.
  
  
  CONCLUSIONS
  The quiescin sulfhydryl oxidase hQSOX1b seems to function as a chaperone and oxidase during the oxidative folding. This example for mFIZZ1 should encourage the design of an appropriate thiol/disulfide oxidoreductase-tuned cell free expression system for other challenging disulfide rich proteins.
authors: Brussels Center for Redox Biology, Brussels, Belgium.
authors: Wael
authors: W
authors: Gad
authors: Meera G
authors: MG
authors: Nair
authors: Karolien
authors: K
authors: Van Belle
authors: Khadija
authors: K
authors: Wahni
authors: Henri
authors: H
authors: De Greve
authors: Jo A
authors: JA
authors: Van Ginderachter
authors: Guy
authors: G
authors: Vandenbussche
authors: Yaeta
authors: Y
authors: Endo
authors: David
authors: D
authors: Artis
authors: Joris
authors: J
authors: Messens
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004220:Disulfides; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005854:Germ Cells / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D012995:Solubility; D014908:Triticum / Q000235:genetics* / Q000378:metabolism
chemical_list: D004220:Disulfides; D036341:Intercellular Signaling Peptides and Proteins; C413576:Retnla protein, mouse; D050862:Oxidoreductases Acting on Sulfur Group Donors; C117183:QSOX1 protein, human
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