Horrell, Sam and Antonyuk, Svetlana V and Eady, Robert R and Hasnain, S Samar and Hough, Michael A and Strange, Richard W (2016) Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal. IUCrJ, 3 (4). pp. 271-281. DOI https://doi.org/10.1107/s205225251600823x
Horrell, Sam and Antonyuk, Svetlana V and Eady, Robert R and Hasnain, S Samar and Hough, Michael A and Strange, Richard W (2016) Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal. IUCrJ, 3 (4). pp. 271-281. DOI https://doi.org/10.1107/s205225251600823x
Horrell, Sam and Antonyuk, Svetlana V and Eady, Robert R and Hasnain, S Samar and Hough, Michael A and Strange, Richard W (2016) Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal. IUCrJ, 3 (4). pp. 271-281. DOI https://doi.org/10.1107/s205225251600823x
Abstract
<jats:p>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<jats:italic>in crystallo</jats:italic>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.</jats:p>
Item Type: | Article |
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Uncontrolled Keywords: | serial crystallography; catalysis; enzyme mechanism; denitrification; copper nitrite reductase; radiation damage; radiolysis; synchrotron radiation; XFEL; MSOX |
Subjects: | Q Science > Q Science (General) Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology |
Divisions: | Faculty of Science and Health Faculty of Science and Health > Life Sciences, School of |
SWORD Depositor: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 29 Jun 2016 15:46 |
Last Modified: | 30 Oct 2024 20:02 |
URI: | http://repository.essex.ac.uk/id/eprint/17089 |
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