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Abstract
Redox metalloproteins, such as high-potential iron sulfur proteins (HiPIP) which possess a single [Fe4S4] cluster, are ubiquitous in biology performing wide range of physiological functions based on oxidation-reduction (redox) reactions. These redox reactions go through different oxidation states—during catalysis—that are guided by distinct conformational changes preceding and following the stimulation by redox reactant thereby enabling the correct function of the system. These structural changes are of highly dynamic in nature which require a sophisticated visualization method such as time-resolved serial femtosecond crystallography (TR-SFX) to study them. Here, we propose to utilize the high repetition rate capability of the European XFEL to collect SFX data with short-pulse duration (≤10 fs) of the Thermochromatium tepidum HiPIP microcrystals in its resting (reduced) and active (oxidized) state as well as collecting TR-SFX data of the oxidation reaction in series time points after mixing microcrystals with non-physiological oxidizing agents (MISC) using a 3D printed mixing-injector fabricated at the SEC group in the European XFEL. This will allow us to understand; (i) the static structural differences between the resting and active states with less or radiation-damage free metal cluster, and (ii) the structural changes that occur during oxidation immediately after mixing with a redox reactant. Completing such tedious experiments with less amount of samples in a single-beam highly depends on the MHz repetition rate capability of the European XFEL.