Reaction intermediates and redox state changes in a blue laccase from Steccherinum ochraceum observed by crystallographic high/low X-ray dose experiments

Journal of Inorganic Biochemistry
Marta FerraroniFabrizio Briganti

Abstract

The crystal structure of a blue laccase from Steccherinum ochraceum has been solved at 2.0Å of resolution using a classic data acquisition from a single crystal. The overall structural features are typical of this class of enzymes, however, distances inside the trinuclear copper cluster are indicative of a reduction of the metal centers induced by free electrons produced during the X-ray data collection. UV-visible spectra collected during the X-ray exposure support the progressive reduction of the metal centers. In order to better detect the reduction progression steps in the trinuclear copper site, a multicrystal data collection strategy based on a systematic spread of the X-ray dose over many crystals has been employed. This approach is based on collecting multicrystal data sets, then combining the slices of the individual data sets experiencing the same radiation dose to obtain composite complete data sets at progressively higher doses. Applying this technique, we have been able to capture sequential frames of the enzyme during the metal centers and molecular oxygen reduction mechanism obtaining a three-dimensional movie of the X-ray-driven catalytic conversion of the molecular oxygen in the active site of laccase: first, t...Continue Reading

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Citations

Sep 18, 2013·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Nina HakulinenJuha Rouvinen
Aug 15, 2015·Journal of Biochemistry·Hirofumi Komori, Yoshiki Higuchi
Jan 15, 2015·Cellular and Molecular Life Sciences : CMLS·N Hakulinen, J Rouvinen
Mar 25, 2015·Journal of Structural Biology·Tian XieGanggang Wang
Mar 5, 2014·Chemical Reviews·Edward I SolomonLi Tian
May 1, 2018·Proteins·Scot Wherland, Israel Pecht
Jan 17, 2014·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Scot WherlandIsrael Pecht

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