TY  - EJOUR
AU  - Kwiatkowksi, Anna
AU  - Caserta, Giorgio
AU  - Schulz, Anne-Christine
AU  - Frielingsdorf, Stefan
AU  - Pelmenschikov, Vladimir
AU  - Weisser, Kilian
AU  - Belsom, Adam
AU  - Rappsilber, Juri
AU  - Sergeev, Ilya
AU  - Limberg, Christian
AU  - Mroginski, Maria-Andrea
AU  - Zebger, Ingo
AU  - Lenz, Oliver
TI  - ATP-triggered Fe(CN)<sub>2</sub>CO synthon transfer from the maturase HypCD to the active site of apo-[NiFe]-hydrogenase
M1  - PUBDB-2024-05723
PY  - 2024
N1  - 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).
AB  - [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. 
LB  - PUB:(DE-HGF)25
DO  - DOI:10.26434/chemrxiv-2024-3dx2g
UR  - https://bib-pubdb1.desy.de/record/614022
ER  -