%0 Electronic Article
%A Kwiatkowksi, Anna
%A Caserta, Giorgio
%A Schulz, Anne-Christine
%A Frielingsdorf, Stefan
%A Pelmenschikov, Vladimir
%A Weisser, Kilian
%A Belsom, Adam
%A Rappsilber, Juri
%A Sergeev, Ilya
%A Limberg, Christian
%A Mroginski, Maria-Andrea
%A Zebger, Ingo
%A Lenz, Oliver
%T ATP-triggered Fe(CN)<sub>2</sub>CO synthon transfer from the maturase HypCD to the active site of apo-[NiFe]-hydrogenase
%M PUBDB-2024-05723
%D 2024
%Z Funding: Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) through SPP 1927 “Iron Sulfur for Life” project no 311062227; the cluster of excellence ‘UniSysCat’ under Germany’s Excellence Strategy-EXC2008-390540038. EU: the COST Action FeSImmChemNet, CA21115, supported by COST (European Cooperation in Science and Technology).
%X [NiFe]-hydrogenases catalyze the reversible activation of H<sub>2</sub> using a unique NiFe(CN)<sub>2</sub>CO metal site, which is assembled by a sophisticated multi-protein machinery. The [4Fe–4S]-cluster-containing HypCD complex, which possesses an ATPase activity with an hitherto unknown function, serves as the hub for the assembly of the Fe(CN)<sub>2</sub>CO sub-fragment. HypCD is also thought to be responsible for the subsequent transfer of the iron fragment to the apo-form of the catalytic hydrogenase subunit, but the underlying mechanism remained unexplored. Here, we performed a thorough spectroscopic characterization of different HypCD preparations using infrared, Mössbauer and NRVS spectroscopy, revealing molecular details of the coordination of the Fe(CN)<sub>2</sub>CO fragment. Moreover, biochemical assays in combination with spectroscopy, AlphaFold structure predictions, protein-ligand docking calculations and crosslinking MS deciphered unexpected mechanistic aspects of the ATP requirement of HypCD, which we found to actually trigger the transfer of the Fe(CN)<sub>2</sub>CO fragment to the apo-hydrogenase. 
%F PUB:(DE-HGF)25
%9 Preprint
%R 10.26434/chemrxiv-2024-3dx2g
%U https://bib-pubdb1.desy.de/record/614022