Japan Partnering Award -XFEL and dynamic heme enzymology
Lead Research Organisation:
University of Leicester
Department Name: Molecular and Cell Biology
Abstract
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Publications
Kwon H
(2021)
XFEL Crystal Structures of Peroxidase Compound II
in Angewandte Chemie International Edition
Kwon H
(2021)
XFEL Crystal Structures of Peroxidase Compound II
in Angewandte Chemie
| Description | By trapping a key intermediate (Compound II) in the catalytic pathway of the enzymes cytochrome c peroxidase and ascorbate peroxidase using the femtosecond pulses of the SACLA X-ray free electron laser to avoid the chemical effects of X-rays, we have shown that there is a key difference in teh distances between the iron and oxygen atom in this state of these enzymes. This means the details of the chemistry needs to be re-evaluated. |
| Exploitation Route | Use of XFELs to avoid seeing X-ray induced reduction of redox-sensitive sates will be applicable to the investigation of many other redox enzymes |
| Sectors | Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Title | Raw XFEL diffraction data from compound II of Ascorbate peroxidase |
| Description | raw X-ray data deposition for publication 10.1002/anie.202103010 and PDB ID 7BI1 |
| Type Of Material | Database/Collection of data |
| Year Produced | 2021 |
| Provided To Others? | Yes |
| Impact | making data publicly available will enable improvement in processing |
| Title | Raw diffraction images of CCP by SF-ROX at SACLA |
| Description | Diffraction images of cytochrome c peroxidase (CcP, PDB code: 7BIU). They were collected using X-ray free electron laser from SACLA at 15 keV with the serial femtosecond rotation crystallography (SF-ROX) method. The MX300-HS CCD detector was used to record images. CcP crystals belong to P212121 space group with a=50.81, b=75.54, c=106.57 Å. 10303 images were collected from 147 crystals, and 9145 images were merged at 1.06 Å resolution using CrystFEL 0.9.1 in the published result. The experiment was carried out on BL2. Images were converted to HDF5 files with measured photon energies. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2021 |
| Provided To Others? | Yes |
| Impact | public availably of data will enable improvement of data processing methods |
| URL | https://zenodo.org/record/4484116 |
| Description | Diamond Light Source |
| Organisation | Diamond Light Source |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Provided material for X-ray crystallography and X-ray fluorescence measurements, conducted or helped to conduct experiments |
| Collaborator Contribution | provided facilities and expert advice (including hands-on help) when requested |
| Impact | Nature Communications article (2016) listed in outputs |
| Start Year | 2014 |
| Description | SACLA - Spring 8 X-ray free electron laser |
| Organisation | RIKEN |
| Department | RIKEN SPring-8 Center |
| Country | Japan |
| Sector | Private |
| PI Contribution | We provided ~150 crystals of the Compound II intermediate of Ascorbate Peroxidase cryo preserved for multiple exposures on large crystals x-ray data collection at the free electron laser |
| Collaborator Contribution | Beam time at the free-electron laser, experimental data collection expertise, preliminary data proecessing |
| Impact | Initial results were presented at the European Crystallographic Meeting in Oviedo, Spain (2018), We have used the data to test and explore the use of the DIALS software (in collaboration with Dr Evan's group at Diamond Light Source) and are comparing the result to that from the preliminary processing with CrystalFEL. Further test are needed to optimise data collection protocols with this method, and we are applying for time to also collect data on CcP Compound II. |
| Start Year | 2018 |
| Description | LINXS Reboot After the pandemics |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Talk given as contribution to meeting "to discuss the LINXS science and development in the past four years and the future ambitions in expanding the LINXS community." Lund Sweden. Establishing contacts with ESS. MAXIV and Lund University community |
| Year(s) Of Engagement Activity | 2022 |
| URL | https://www.linxs.se/events/2022/2/2/linxs-science-day-the-development-and-future-vision-of-linxs |