Horrell, S, Antonyuk, SV 
ORCID: 0000-0002-2779-9946, Eady, RR, Hasnain, SS, Hough, MA and Strange, RW
(2016)
Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal.
IUCrJ, 3 (Part 4).
pp. 271-281.
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Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal.pdf - Published version Download (2MB) |
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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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | serial crystallography, catalysis, enzyme mechanism, denitrification, copper nitrite reductase, radiation damage, radiolysis, synchrotron radiation, XFEL, MSOX |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 26 Oct 2016 15:19 |
| Last Modified: | 19 Jan 2023 07:27 |
| DOI: | 10.1107/S205225251600823X |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3004102 |
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