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Structure of the sulphiredoxin–peroxiredoxin complex reveals an essential repair embrace

Nature volume 451, pages 98–101 (2008)Cite this article

Abstract

Typical 2-Cys peroxiredoxins (Prxs) have an important role in regulating hydrogen peroxide-mediated cell signalling1. In this process, Prxs can become inactivated through the hyperoxidation of an active site Cys residue to Cys sulphinic acid. The unique repair of this moiety by sulphiredoxin (Srx) restores peroxidase activity and terminates the signal2. The hyperoxidized form of Prx exists as a stable decameric structure with each active site buried. Therefore, it is unclear how Srx can access the sulphinic acid moiety. Here we present the 2.6 Å crystal structure of the human Srx–PrxI complex. This complex reveals the complete unfolding of the carboxy terminus of Prx, and its unexpected packing onto the backside of Srx away from the Srx active site. Binding studies and activity analyses of site-directed mutants at this interface show that the interaction is required for repair to occur. Moreover, rearrangements in the Prx active site lead to a juxtaposition of the Prx Gly-Gly-Leu-Gly and Srx ATP-binding motifs, providing a structural basis for the first step of the catalytic mechanism. The results also suggest that the observed interactions may represent a common mode for other proteins to bind to Prxs.

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Figure 1: Peroxiredoxin hyperoxidation and repair by sulphiredoxin.
Figure 2: Srx-PrxI active site interactions and structural plasticity.
Figure 3: PrxI backside interaction with Srx.

Accession codes

Primary accessions

Protein Data Bank

Data deposits

Coordinates and structure factors have been deposited with the Protein Data Bank under the accession number 2RII.

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Acknowledgements

We thank M. Murray for contributions to the structure determination, L. B. Poole, P. A. Karplus and N. H. Heintz for discussions, the staff of the NSLS and beamline X8C for their assistance during data collection and the RapiData course, and R. R. Hantgan for help with the circular dichroism and fluorescence anisotropy experiments. This work was supported by an NIH grant (W.T.L.) and an American Heart Association Postdoctoral Fellowship (T.J.J.). NSLS is supported by the US Department of Energy and NIH.

Author Contributions T.J.J. and L.C.J. performed all biochemical and crystallization experiments. T.J.J. and W.T.L solved the structure. T.J.J. and W.T.L. wrote the paper. All authors discussed the results and commented on the manuscript.

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Authors and Affiliations

  1. Center for Structural Biology and Department of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, North Carolina 27157, USA,

    Thomas J. Jönsson, Lynnette C. Johnson & W. Todd Lowther

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  1. Thomas J. Jönsson
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  2. Lynnette C. Johnson
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  3. W. Todd Lowther
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Correspondence to W. Todd Lowther.

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The file contains Supplementary Table S1, Supplementary Figures S1-S9 with Legends and additional references. (PDF 713 kb)

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Jönsson, T., Johnson, L. & Lowther, W. Structure of the sulphiredoxin–peroxiredoxin complex reveals an essential repair embrace. Nature 451, 98–101 (2008). https://doi.org/10.1038/nature06415

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