Reconstitution of a protein disulfide catalytic system.

The Journal of Biological Chemistry
Martin W BaderJ C Bardwell

Abstract

Disulfide bonds are important for the structure and stability of many proteins. In prokaryotes their formation is catalyzed by the Dsb proteins. The DsbA protein acts as a direct donor of disulfides to newly synthesized periplasmic proteins. Genetic evidence suggests that a second protein called DsbB acts to specifically reoxidize DsbA. Here we demonstrate the direct reoxidation of DsbA by DsbB. We have developed a fluorescence assay that allows us to directly follow the reoxidation of DsbA. We show that membranes containing catalytic amounts of DsbB can rapidly reoxidize DsbA to completion. The reaction strongly depends on the presence of oxygen, implying that oxygen serves as the final electron acceptor for this disulfide bond formation reaction. Membranes from a dsbB null mutant display no DsbA reoxidation activity. The ability of DsbB to reoxidize DsbA fits Michaelis-Menten behavior with DsbA acting as a high affinity substrate for DsbB with a Km = 10 microM. The in vitro reconstitution described here is the first biochemical analysis of DsbB and allows us to study the major pathway of disulfide bond formation in Escherichia coli.

References

Nov 1, 1989·Analytical Biochemistry·S C Gill, P H von Hippel
Oct 10, 1995·Proceedings of the National Academy of Sciences of the United States of America·C GuilhotJ Beckwith
Aug 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·D MissiakasS Raina
Feb 1, 1995·Current Opinion in Structural Biology·R B Freedman
Oct 1, 1994·Molecular Microbiology·J C Bardwell
Mar 17, 1995·Journal of Molecular Biology·M WunderlichR Glockshuber
Feb 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·J C BardwellJ Beckwith
May 1, 1993·Protein Science : a Publication of the Protein Society·M Wunderlich, R Glockshuber
Dec 15, 1995·Cell·U GrauschopfJ C Bardwell
Feb 1, 1996·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·S Kishigami, K Ito
Oct 29, 1997·Proceedings of the National Academy of Sciences of the United States of America·T KobayashiK Ito

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Citations

Aug 5, 2003·BioFactors·Gábor BánhegyiJózsef Mandl
Sep 24, 2005·Archives of Microbiology·Alexis DumoulinBrigitte Berger-Bächi
Jun 8, 2000·FEMS Microbiology Reviews·R A FabianekL Thöny-Meyer
Jan 30, 2010·Nature·Weikai LiTom A Rapoport
Apr 23, 2002·Molecular Microbiology·Jean-Francois Collet, James C A Bardwell
May 21, 2010·Proceedings of the National Academy of Sciences of the United States of America·Kalle MöbiusDieter Jahn
Sep 19, 2003·Antioxidants & Redox Signaling·Ron Ortenberg, Jon Beckwith
Jun 15, 2006·Antioxidants & Redox Signaling·Yoh-Hei TakahashiKoreaki Ito
Mar 11, 2010·Antioxidants & Redox Signaling·Dionisia P Sideris, Kostas Tokatlidis
May 30, 2009·Molecular Biology of the Cell·Heather L TiensonCarla M Koehler
Sep 21, 2012·Acta Crystallographica. Section D, Biological Crystallography·Patricia M WaldenJennifer L Martin
Jun 19, 2002·Protein Science : a Publication of the Protein Society·Floriana VinciMireille Moutiez
Feb 3, 2005·Journal of Bacteriology·Jacqueline TanJames C A Bardwell
Dec 12, 2013·MBio·Isabelle S ArtsJean-François Collet
Sep 27, 2000·Proceedings of the National Academy of Sciences of the United States of America·H KadokuraJ Beckwith
Aug 9, 2001·Proceedings of the National Academy of Sciences of the United States of America·D GoldstoneP Metcalf
Nov 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·James RegeimbalJames C A Bardwell
Aug 16, 2005·Progress in Biophysics and Molecular Biology·Alexandra P CarvalhoMaria J Ramos
Nov 18, 2004·Biochimica Et Biophysica Acta·Hitoshi Nakamoto, James C A Bardwell
Mar 18, 2008·Biochimica Et Biophysica Acta·Stefan Gleiter, James C A Bardwell
Sep 7, 2001·Annual Review of Microbiology·D Ritz, J Beckwith
Oct 10, 2009·Journal of Molecular Biology·Julian P VivianMartin J Scanlon
Mar 13, 1999·The Journal of Biological Chemistry·P H BessetteG Georgiou
Nov 13, 1999·Nature Cell Biology·M BaderJ C Bardwell
Sep 15, 1999·Nature·R Glockshuber
Mar 3, 1999·Nature Biotechnology·M M AltamiranoA R Fersht
Jun 25, 2004·The EMBO Journal·Yoshiaki FurukawaThomas V O'Halloran
Aug 24, 2002·The Journal of Biological Chemistry·Bjorn Philipps, Rudi Glockshuber
Oct 30, 2004·Chembiochem : a European Journal of Chemical Biology·Jacqueline T Tan, James C A Bardwell
Nov 5, 2002·Nature Reviews. Molecular Cell Biology·Carolyn S Sevier, Chris A Kaiser
Feb 3, 2007·The Journal of Biological Chemistry·Timothy L TapleyJames C A Bardwell
Aug 22, 2008·The Journal of Biological Chemistry·Julian P VivianMartin J Scanlon

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