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A cryo-crystallographic time course for peroxide reduction by rubrerythrin from Pyrococcus furiosus

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Abstract

High-resolution crystal structures of Pyrococcus furiosus rubrerythrin (PfRbr) in the resting (all-ferrous) state and at time points following exposure of the crystals to hydrogen peroxide are reported. This approach was possible because of the relativity slow turnover of PfRbr at room temperature. To this end, we were able to perform time-dependent peroxide treatment of the fully reduced enzyme, under strictly anaerobic conditions, in the crystalline state. In this work we demonstrate, for the first time, that turnover of a thermophilic rubrerythrin results in approximately 2-Å movement of one iron atom in the diiron site from a histidine to a carboxylate ligand. These results confirm that, despite the domain-swapped architecture, the hyperthermophilic rubrerythrins also utilize the classic combination of iron sites together with redox-dependent iron toggling to selectively reduce hydrogen peroxide over dioxygen. In addition, we have identified previously unobserved intermediates in the reaction cycle and observed structural changes that may explain the enzyme precipitation observed for the all-iron form of PfRbr upon oxidation to the all-ferric state.

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Acknowledgments

This research was supported by NSF grant MCB 0835432 to W.N.L. and a grant (DE-FG05-95ER20175) to M.W.A. from the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, US Department of Energy.

Conflict of interest

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The Center for Metalloenzyme Studies, University of Georgia, Athens, GA, 30602, USA

    Bret D. Dillard, Jonathan M. Demick, Michael W. W. Adams & William N. Lanzilotta

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  1. Bret D. Dillard
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  2. Jonathan M. Demick
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  3. Michael W. W. Adams
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  4. William N. Lanzilotta
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Corresponding author

Correspondence to William N. Lanzilotta.

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An interactive 3D complement page in Proteopedia is available at http://proteopedia.org/wiki/index.php/Journal:JBIC-11-02-00042.

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Dillard, B.D., Demick, J.M., Adams, M.W.W. et al. A cryo-crystallographic time course for peroxide reduction by rubrerythrin from Pyrococcus furiosus . J Biol Inorg Chem 16, 949–959 (2011). https://doi.org/10.1007/s00775-011-0795-6

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  • DOI: https://doi.org/10.1007/s00775-011-0795-6

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