Experimentally Dissecting the Origins of Peroxiredoxin Catalysis.

Journal Article (Journal Article)


Peroxiredoxins (Prxs) are ubiquitous cysteine-based peroxidases involved in oxidant defense and signal transduction. Despite much study, the precise roles of conserved residues remain poorly defined. In this study, we carried out extensive functional and structural characterization of 10 variants of such residues in a model decameric bacterial Prx.


Three active site proximal mutations of Salmonella typhimurium AhpC, T43V, R119A, and E49Q, lowered catalytic efficiency with hydrogen peroxide by 4-5 orders of magnitude, but did not affect reactivity toward their reductant, AhpF. pKa values of the peroxidatic cysteine were also shifted up by 1-1.3 pH units for these and a decamer disruption mutant, T77I. Except for the decamer-stabilizing T77V, all mutations destabilized decamers in the reduced form. In the oxidized form, three mutants-T77V, T43A, and T43S-exhibited stabilized decamers and were more efficiently reduced by AhpF than wild-type AhpC. Crystal structures of most mutants were solved and many showed alterations in stability of the fully folded active site loop.


This is the first study of Prx mutants to comprehensively assess the effects of mutations on catalytic activities, the active site cysteine pKa , and the protein structure and oligomeric status.


The Arg119 side chain must be properly situated for efficient catalysis, but for other debilitating variants, the functional defects could be explained by structural perturbations and/or associated decamer destabilization rather than direct effects. This underscores the importance of our comprehensive approach. A remarkable new finding was the preference of the reductant for decamers. Antioxid. Redox Signal. 28, 521-536.

Full Text

Duke Authors

Cited Authors

  • Nelson, KJ; Perkins, A; Van Swearingen, AED; Hartman, S; Brereton, AE; Parsonage, D; Salsbury, FR; Karplus, PA; Poole, LB

Published Date

  • March 2018

Published In

Volume / Issue

  • 28 / 7

Start / End Page

  • 521 - 536

PubMed ID

  • 28375740

Pubmed Central ID

  • PMC5806077

Electronic International Standard Serial Number (EISSN)

  • 1557-7716

International Standard Serial Number (ISSN)

  • 1523-0864

Digital Object Identifier (DOI)

  • 10.1089/ars.2016.6922


  • eng