Control of periplasmic interdomain thiol:disulfide exchange in the transmembrane oxidoreductase DsbD

Mavridou, Despoina A. I. and Stevens, Julie M. and Goddard, Alan D. and Willis, Anthony C. and Ferguson, Stuart J. and Redfield, Christina (2009) Control of periplasmic interdomain thiol:disulfide exchange in the transmembrane oxidoreductase DsbD. Journal of Biological Chemistry, 284 (5). pp. 3219-3226. ISSN 0021-9258

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Official URL: http://dx.doi.org/10.1074/jbc.M805963200

Abstract

The bacterial protein DsbD transfers reductant from the cytoplasm to the otherwise oxidizing environment of the periplasm. This reducing power is required for several essential pathways, including disulfide bond formation and cytochrome c maturation. DsbD includes a transmembrane domain (tmDsbD) flanked by two globular periplasmic domains (nDsbD/cDsbD); each contains a cysteine pair involved in electron transfer via a disulfide exchange cascade. The final step in the cascade involves reduction of the Cys 103-Cys 109 disulfide of nDsbD by Cys 461 of cDsbD. Here we show that a complex between the globular periplasmic domains is trapped in vivo only when both are linked by tmDsbD. We have found previously (Mavridou, D. A., Stevens, J. M., Ferguson, S. J., & Redfield, C. (2007) J. Mol. Biol. 370, 643-658) that the attacking cysteine (Cys 461) in isolated cDsbD has a high pK a value (10.5) that makes this thiol relatively unreactive toward the target disulfide in nDsbD. Here we show using NMR that active-site pK a values change significantly when cDsbD forms a complex with nDsbD. This modulation of pK a values is critical for the specificity and function of cDsbD. Uncomplexed cDsbD is a poor nucleophile, allowing it to avoid nonspecific reoxidation; however, in complex with nDsbD, the nucleophilicity of cDsbD increases permitting reductant transfer. The observation of significant changes in active-site pK a values upon complex formation has wider implications for understanding reactivity in thiol:disulfide oxidoreductases.

Item Type:Article
Keywords:DsbD
Subjects:C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
ID Code:6832
Deposited By: Alan Goddard
Deposited On:15 Nov 2012 21:31
Last Modified:15 Nov 2013 17:00

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