Abstract
Heme peroxidases catalyze the H2O2-dependent oxidation of a variety of substrates, most of which are organic. Mechanistically, these enzymes are well characterized: they share a common catalytic cycle that involves formation of a two-electron, oxidized Compound I intermediate followed by two single-electron reduction steps by substrate. The substrate specificity is more diverse — most peroxidases oxidize small organic substrates, but there are prominent exceptions — and there is a notable absence of structural information for a representative peroxidase–substrate complex. Thus, the features that control substrate specificity remain undefined. We present the structure of the complex of ascorbate peroxidase–ascorbate. The structure defines the ascorbate-binding interaction for the first time and provides new rationalization of the unusual functional features of the related cytochrome c peroxidase enzyme, which has been a benchmark for peroxidase catalysis for more than 20 years. A new mechanism for electron transfer is proposed that challenges existing views of substrate oxidation in other peroxidases.
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Acknowledgements
We are grateful to D. Dalton for providing the expression vector for rsAPX. We thank K. Singh for technical assistance, D. Leys for data collection and C. Metcalfe for helpful discussions. We are grateful to the EMBL outstation at DESY Hamburg and ESRF Grenoble for the provision of synchrotron radiation. This work was supported by grants from the BBSRC, the Wellcome Trust, the EPSRC and the Royal Society.
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Sharp, K., Mewies, M., Moody, P. et al. Crystal structure of the ascorbate peroxidase–ascorbate complex. Nat Struct Mol Biol 10, 303–307 (2003). https://doi.org/10.1038/nsb913
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DOI: https://doi.org/10.1038/nsb913
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