Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal
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 c...
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| Format: | Article |
| Language: | English |
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International Union of Crystallography
2016-07-01
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| Series: | IUCrJ |
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| Online Access: | http://scripts.iucr.org/cgi-bin/paper?S205225251600823X |
| _version_ | 1818321990793035776 |
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| author | Sam Horrell Svetlana V. Antonyuk Robert R. Eady S. Samar Hasnain Michael A. Hough Richard W. Strange |
| author_facet | Sam Horrell Svetlana V. Antonyuk Robert R. Eady S. Samar Hasnain Michael A. Hough Richard W. Strange |
| author_sort | Sam Horrell |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T10:49:41Z |
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| id | doaj.art-434ddd5033b24b76b98c35d07d3ccdb2 |
| institution | Directory Open Access Journal |
| issn | 2052-2525 |
| language | English |
| last_indexed | 2024-12-13T10:49:41Z |
| publishDate | 2016-07-01 |
| publisher | International Union of Crystallography |
| record_format | Article |
| series | IUCrJ |
| spelling | doaj.art-434ddd5033b24b76b98c35d07d3ccdb22022-12-21T23:49:55ZengInternational Union of CrystallographyIUCrJ2052-25252016-07-013427128110.1107/S205225251600823Xlz5011Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystalSam Horrell0Svetlana V. Antonyuk1Robert R. Eady2S. Samar Hasnain3Michael A. Hough4Richard W. Strange5School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, EnglandMolecular Biophysics Group, Institute of Integrative Biology, University of Liverpool, Life Sciences Building, Liverpool L69 7ZB, EnglandMolecular Biophysics Group, Institute of Integrative Biology, University of Liverpool, Life Sciences Building, Liverpool L69 7ZB, EnglandMolecular Biophysics Group, Institute of Integrative Biology, University of Liverpool, Life Sciences Building, Liverpool L69 7ZB, EnglandSchool of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, EnglandSchool of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, EnglandRelating 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.http://scripts.iucr.org/cgi-bin/paper?S205225251600823Xserial crystallographycatalysisenzyme mechanismdenitrificationcopper nitrite reductaseradiation damageradiolysissynchrotron radiationXFELMSOX |
| spellingShingle | Sam Horrell Svetlana V. Antonyuk Robert R. Eady S. Samar Hasnain Michael A. Hough Richard W. Strange Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal IUCrJ serial crystallography catalysis enzyme mechanism denitrification copper nitrite reductase radiation damage radiolysis synchrotron radiation XFEL MSOX |
| title | Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| title_full | Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| title_fullStr | Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| title_full_unstemmed | Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| title_short | Serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| title_sort | serial crystallography captures enzyme catalysis in copper nitrite reductase at atomic resolution from one crystal |
| topic | serial crystallography catalysis enzyme mechanism denitrification copper nitrite reductase radiation damage radiolysis synchrotron radiation XFEL MSOX |
| url | http://scripts.iucr.org/cgi-bin/paper?S205225251600823X |
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