Engineered DsbC chimeras catalyze both protein oxidation and disulfide-bond isomerization in Escherichia coli: Reconciling two competing pathways

Proceedings of the National Academy of Sciences of the United States of America
Laura SegatoriG Georgiou

Abstract

In the Escherichia coli periplasm, the formation of protein disulfide bonds is catalyzed by DsbA and DsbC. DsbA is a monomer that is maintained in a fully oxidized state by the membrane enzyme DsbB, whereas DsbC is a dimer that is kept reduced by a second membrane protein, DsbD. Although the catalytic regions of DsbA and DsbC are composed of structurally homologous thioredoxin motif domains, DsbA serves only as an oxidase in vivo, whereas DsbC catalyzes disulfide reduction and isomerization and also exhibits significant chaperone activity. To reconcile the distinct catalytic activities of DsbC and DsbA, we constructed a series of chimeras comprising of the dimerization domain of DsbC, with or without the adjacent alpha-helical linker region, fused either to the first, second, third, or fifth residue of intact DsbA or to thioredoxin. The chimeras fully substituted for DsbC in disulfide-bond rearrangement and also were able to restore protein oxidation in a dsbA background. Remarkably, the chimeras could serve as a single catalyst for both disulfide-bond formation and rearrangement, thus reconciling the kinetically competing DsbB-DsbA and DsbD-DsbC pathways. This property appeared to depend on the orientation of the DsbA active-s...Continue Reading

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Citations

Jul 8, 2009·The Journal of Biological Chemistry·Silvia A ArredondoGeorge Georgiou
Sep 11, 2008·The Journal of Biological Chemistry·Silvia ArredondoGeorge Georgiou
Apr 7, 2010·Antioxidants & Redox Signaling·Hiroshi Kadokura, Jon Beckwith
Sep 21, 2010·Antioxidants & Redox Signaling·Matthieu DepuydtJean-Francois Collet
Mar 1, 2014·Biochimica Et Biophysica Acta·Feras HatahetJon Beckwith
May 15, 2007·Proceedings of the National Academy of Sciences of the United States of America·Ki Jun JeongGeorge Georgiou
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Feb 13, 2021·Molecular Microbiology·Sabrina L Slater, Despoina A I Mavridou
Feb 15, 2019·EcoSal Plus·Bruno MantaMehmet Berkmen

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