Japan Partnering Award -XFEL and dynamic heme enzymology

Lead Research Organisation: University of Leicester
Department Name: Molecular and Cell Biology


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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

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