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Supplementary information files for Application of a synthetic ferredoxin‐inspired [4Fe4S]‐peptide maquette as the redox partner for a [FeFe]‐hydrogenase

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Version 2 2023-08-14, 16:00
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posted on 2023-08-14, 16:00 authored by Andrea Bombana, Muralidharan Shanmugam, David Collison, Alexander J Kibler, Graham N Newton, Christof M Jager, Anna CroftAnna Croft, Simone K Morra, Nicholas Mitchell

Supplementrary files for article Application of a synthetic ferredoxin‐inspired [4Fe4S]‐peptide maquette as the redox partner for a [FeFe]‐hydrogenase

‘Bacterial-type’ ferredoxins host a cubane [4Fe4S]2+/+ cluster that enables these proteins to mediate electron transfer and facilitate a broad range of biological processes. Peptide maquettes based on the conserved cluster-forming motif have previously been reported and used to model the ferredoxins. Herein we explore the integration of a [4Fe4S]-peptide maquette into a H2-powered electron transport chain. While routinely formed under anaerobic conditions, we illustrate by electron paramagnetic resonance (EPR) analysis that these maquettes can be reconstituted under aerobic conditions by using photoactivated NADH to reduce the cluster at 240 K. Attempts to tune the redox properties of the iron-sulfur cluster by introducing an Fe-coordinating selenocysteine residue were also explored. To demonstrate the integration of these artificial metalloproteins into a semi-synthetic electron transport chain, we utilize a ferredoxin-inspired [4Fe4S]-peptide maquette as the redox partner in the hydrogenase-mediated oxidation of H2. 

Funding

Site-selective functionalisation of peptides and proteins via free-radical-induced dechalcogenation

Engineering and Physical Sciences Research Council

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Equipment to support Nottingham's Research Fellows

Engineering and Physical Sciences Research Council

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EPSRC National Electron Paramagnetic Resonance Facility and Service

Engineering and Physical Sciences Research Council

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A National Research Facility for EPR Spectroscopy, 2022-2027

Engineering and Physical Sciences Research Council

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