Direct evidence for a peroxide intermediate and a reactive enzyme-substrate-dioxygen configuration in a cofactor-free oxidase.

Abstract : Cofactor-free oxidases and oxygenases promote and control the reactivity of O2 with limited chemical tools at their disposal. Their mechanism of action is not completely understood and structural information is not available for any of the reaction intermediates. Near-atomic resolution crystallography supported by in crystallo Raman spectroscopy and QM/MM calculations showed unambiguously that the archetypical cofactor-free uricase catalyzes uric acid degradation via a C5(S)-(hydro)peroxide intermediate. Low X-ray doses break specifically the intermediate C5-OO(H) bond at 100 K, thus releasing O2 in situ, which is trapped above the substrate radical. The dose-dependent rate of bond rupture followed by combined crystallographic and Raman analysis indicates that ionizing radiation kick-starts both peroxide decomposition and its regeneration. Peroxidation can be explained by a mechanism in which the substrate radical recombines with superoxide transiently produced in the active site.
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Article dans une revue
Angewandte Chemie (English Edition), John Wiley & Sons, 2014, 53 (50), pp.13710-4
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http://hal.univ-grenoble-alpes.fr/hal-01148970
Contributeur : Frank Thomas <>
Soumis le : mercredi 6 mai 2015 - 09:30:20
Dernière modification le : lundi 23 avril 2018 - 14:45:34

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  • HAL Id : hal-01148970, version 1
  • PUBMED : 25314114

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Soi Bui, David Von Stetten, Pablo G Jambrina, Thierry Prangé, Nathalie Colloc'H, et al.. Direct evidence for a peroxide intermediate and a reactive enzyme-substrate-dioxygen configuration in a cofactor-free oxidase.. Angewandte Chemie (English Edition), John Wiley & Sons, 2014, 53 (50), pp.13710-4. 〈hal-01148970〉

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