Preprint

PUBDB-2024-05723

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png ATP-triggered Fe(CN)$_2$CO synthon transfer from the maturase HypCD to the active site of apo-[NiFe]-hydrogenase

Kwiatkowksi, A. (First author) ; Caserta, G. (Corresponding author) ; Schulz, A.-C. ; Frielingsdorf, S. ; Pelmenschikov, V. ; Weisser, K. ; Belsom, A. ; Rappsilber, J. ; Sergeev, I.XFEL.EU*DESY* ; Limberg, C. ; Mroginski, M.-A. ; Zebger, I. ; Lenz, O. (Corresponding author)

2024

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Please use a persistent id in citations: doi:10.26434/chemrxiv-2024-3dx2g  doi:10.3204/PUBDB-2024-05723

Abstract: [NiFe]-hydrogenases catalyze the reversible activation of H$_2$ using a unique NiFe(CN)$_2$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)$_2$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)$_2$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)$_2$CO fragment to the apo-hydrogenase.

Note: 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).

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Contributing Institute(s):

1. DOOR-User (DOOR ; HAS-User)
2. Experimentebetreuung PETRA III (FS-PET-S)

Research Program(s):

1. 632 - Materials – Quantum, Complex and Functional Materials (POF4-632) (POF4-632)
2. 6G3 - PETRA III (DESY) (POF4-6G3) (POF4-6G3)
3. FS-Proposal: I-20220851 (I-20220851) (I-20220851)
4. FS-Proposal: I-20210325 (I-20210325) (I-20210325)
5. DFG project G:(GEPRIS)311062227 - Die Rolle von Eisen-Schwefel-Kofaktoren in der Assemblierung von Metallzentren und der Katalyse von Hydrogenasen (311062227) (311062227)
6. DFG project G:(GEPRIS)449713269 - Design, Synthese und Analyse von photospaltbaren chemischen Crosslinkern zur Entwicklung von verbesserten Methoden in der Crosslinking Massenspektrometrie (449713269) (449713269)

Experiment(s):

1. PETRA Beamline P01 (PETRA III)

Appears in the scientific report 2024

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