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• W4385859427 abstract "The ultrashort (10s of femtoseconds) X-ray pulses generated by X-ray free electron lasers enable the measurement of X-ray diffraction and spectroscopic data from radiation-sensitive metalloenzymes at room temperature while mostly avoiding the effects of radiation damage usually encountered when performing such experiments at synchrotron sources. Here we discuss an approach to measure both X-ray emission and X-ray crystallographic data at the same time from the same sample volume. The droplet-on-tape setup described allows for efficient sample use and the integration of different reaction triggering options in order to conduct time-resolved studies with limited sample amounts. The approach is illustrated by two examples, photosystem II that catalyzes the light-driven oxidation of water to oxygen, and isopenicillin N synthase, an enzyme that catalyzes the double ring cyclization of a tripeptide precursor into the β-lactam isopenicillin and can be activated by oxygen exposure. We describe the necessary steps to obtain microcrystals of both proteins as well as the operation procedure for the drop-on-tape setup and details of the data acquisition and processing involved in this experiment. At the end, we present how the combination of time-resolved X-ray emission spectra and diffraction data can be used to improve the knowledge about the enzyme reaction mechanism." @default.
• W4385859427 created "2023-08-17" @default.
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• W4385859427 date "2023-01-01" @default.
• W4385859427 modified "2023-10-17" @default.
• W4385859427 title "Room temperature crystallography and X-ray spectroscopy of metalloenzymes" @default.
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• W4385859427 doi "https://doi.org/10.1016/bs.mie.2023.07.009" @default.
• W4385859427 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/37748830" @default.
• W4385859427 hasPublicationYear "2023" @default.
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