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Direct evidence for a peroxide intermediate and a reactive enzyme–substrate–dioxygen configuration in a cofactor-free oxidase
DOI:
10.1002/anie.201405485
PMID:
25314114
Authors:
Soi
Bui
(King's College London)
,
David
Von stetten
(ESRF)
,
Pablo G.
Jambrina
(King's College London)
,
Thierry
Prangé
(Université Paris Descartes-CNRS)
,
Nathalie
Colloc'H
(Normandie Université)
,
Daniele
De sanctis
(ESRF)
,
Antoine
Royant
(ESRF)
,
Edina
Rosta
(King's College London)
,
Roberto A.
Steiner
(King's College London)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Angewandte Chemie International Edition
, VOL 53 (50)
, PAGES 13710 - 13714
State:
Published (Approved)
Published:
December 2014
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 C5OO(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.
Diamond Keywords: Enzymes
Subject Areas:
Biology and Bio-materials,
Chemistry
Instruments:
I02-Macromolecular Crystallography
Added On:
18/11/2015 14:30
Discipline Tags:
Biochemistry
Chemistry
Structural biology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)