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Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal

DOI: 10.1107/S205225251600823X DOI Help

Authors: Sam Horrell (University of Essex) , Svetlana V. Antonyuk (University of Liverpool) , Robert R. Eady (University of Liverpool) , S. Samar Hasnain (University of Essex) , Michael A. Hough (University of Essex) , Richard W. Strange (University of Essex)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 3 , PAGES 271 - 281

State: Published (Approved)
Published: July 2016
Diamond Proposal Number(s): 11740

Open Access Open Access

Abstract: Relating individual protein crystal structures to an enzyme mechanism remains a major and challenging goal for structural biology. Serial crystallography using multiple crystals has recently been reported in both synchrotron-radiation and X-ray free-electron laser experiments. In this work, serial crystallography was used to obtain multiple structures serially from one crystal (MSOX) to study in crystallo enzyme catalysis. Rapid, shutterless X-ray detector technology on a synchrotron MX beamline was exploited to perform low-dose serial crystallography on a single copper nitrite reductase crystal, which survived long enough for 45 consecutive 100 K X-ray structures to be collected at 1.07–1.62 Å resolution, all sampled from the same crystal volume. This serial crystallography approach revealed the gradual conversion of the substrate bound at the catalytic type 2 Cu centre from nitrite to nitric oxide, following reduction of the type 1 Cu electron-transfer centre by X-ray-generated solvated electrons. Significant, well defined structural rearrangements in the active site are evident in the series as the enzyme moves through its catalytic cycle, namely nitrite reduction, which is a vital step in the global denitrification process. It is proposed that such a serial crystallography approach is widely applicable for studying any redox or electron-driven enzyme reactions from a single protein crystal. It can provide a `catalytic reaction movie' highlighting the structural changes that occur during enzyme catalysis. The anticipated developments in the automation of data analysis and modelling are likely to allow seamless and near-real-time analysis of such data on-site at some of the powerful synchrotron crystallographic beamlines.

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I04-Macromolecular Crystallography

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