|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.|
|Publication Type||Journal Article|
|Year of Publication||2013|
|Authors||Gad, W., M. G. Nair, K. Van Belle, K. Wahni, H. De Greve, J. A. Van Ginderachter, G. Vandenbussche, Y. Endo, D. Artis, and J. Messens|
|Keywords||Amino Acid Sequence, Animals, Disulfides, Escherichia coli, Gene Expression Regulation, Germ Cells, Humans, Intercellular Signaling Peptides and Proteins, Mice, Molecular Sequence Data, Oxidation-Reduction, Oxidoreductases Acting on Sulfur Group Donors, Protein Folding, Protein Structure, Secondary, Sequence Alignment, Solubility, Triticum|
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.
|Alternate Journal||PLoS ONE|
|PubMed Central ID||PMC3561318|
|Grant List||R01 AI091759 / AI / NIAID NIH HHS / United States |
AI061570 / AI / NIAID NIH HHS / United States
AI074878 / AI / NIAID NIH HHS / United States
AI091759 / AI / NIAID NIH HHS / United States
The quiescin sulfhydryl oxidase (hQSOX1b) tunes the expression of resistin-like molecule alpha (RELM-α or mFIZZ1) in a wheat germ cell-free extract.