Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

Proteins belonging to the thioredoxin (Trx) superfamily are abundant in all organisms. They share the same structural features, arranged in a seemingly simple fold, but they perform a multitude of functions in oxidative protein folding and electron transfer pathways. We use the C-terminal domain of the unique transmembrane reductant conductor DsbD as a model for an in-depth analysis of the factors controlling the reactivity of the Trx fold. We employ NMR spectroscopy, x-ray crystallography, mutagenesis, in vivo functional experiments applied to DsbD, and a comparative sequence analysis of Trx-fold proteins to determine the effect of residues in the vicinity of the active site on the ionization of the key nucleophilic cysteine of the -CXXC- motif. We show that the function and reactivity of Trx-fold proteins depend critically on the electrostatic features imposed by an extended active-site motif.

Original publication

DOI

10.1074/jbc.M113.513457

Type

Journal article

Journal

J Biol Chem

Publication Date

21/03/2014

Volume

289

Pages

8681 - 8696

Keywords

DsbD, Electrostatics, NMR, Oxidase, Reductase, Thiol-Disulfide Exchange, Thioredoxin, Thioredoxin Fold, X-ray Crystallography, pKa Values, Amino Acid Motifs, Amino Acid Sequence, Catalytic Domain, Crystallography, X-Ray, Escherichia coli, Escherichia coli Proteins, Humans, Models, Molecular, Molecular Sequence Data, Oxidoreductases, Point Mutation, Protein Structure, Tertiary, Sequence Alignment, Thioredoxins