We found a match
Your institution may have rights to this item. Sign in to continue.
- 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.
- Authors
Gad, Wael; Nair, Meera G.; Van Belle, Karolien; Wahni, Khadija; De Greve, Henri; Van Ginderachter, Jo A.; Vandenbussche, Guy; Endo, Yaeta; Artis, David; Messens, Joris
- 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. Methodology/Principal findings: 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. Conclusion/Significance: 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.
- Subjects
PROTEIN research; DISULFIDES; RECOMBINANT proteins; ESCHERICHIA coli; WHEAT germ; ENZYMES
- Publication
PLoS ONE, 2013, Vol 8, Issue 1, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0055621