Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation

Biochemistry
Eric M ShepardJoan B Broderick

Abstract

Nature utilizes [FeFe]-hydrogenase enzymes to catalyze the interconversion between H2 and protons and electrons. Catalysis occurs at the H-cluster, a carbon monoxide-, cyanide-, and dithiomethylamine-coordinated 2Fe subcluster bridged via a cysteine to a [4Fe-4S] cluster. Biosynthesis of this unique metallocofactor is accomplished by three maturase enzymes denoted HydE, HydF, and HydG. HydE and HydG belong to the radical S-adenosylmethionine superfamily of enzymes and synthesize the nonprotein ligands of the H-cluster. These enzymes interact with HydF, a GTPase that acts as a scaffold or carrier protein during 2Fe subcluster assembly. Prior characterization of HydF demonstrated the protein exists in both dimeric and tetrameric states and coordinates both [4Fe-4S]2+/+ and [2Fe-2S]2+/+ clusters [Shepard, E. M., Byer, A. S., Betz, J. N., Peters, J. W., and Broderick, J. B. (2016) Biochemistry 55, 3514-3527]. Herein, electron paramagnetic resonance (EPR) is utilized to characterize the [2Fe-2S]+ and [4Fe-4S]+ clusters bound to HydF. Examination of spin relaxation times using pulsed EPR in HydF samples exhibiting both [4Fe-4S]+ and [2Fe-2S]+ cluster EPR signals supports a model in which the two cluster types either are bound to wide...Continue Reading

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Citations

Jul 3, 2018·Dalton Transactions : an International Journal of Inorganic Chemistry·Anna G ScottJoan B Broderick
Jul 19, 2018·Metallomics : Integrated Biometal Science·Y BaiA Sawyer
Sep 6, 2019·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Amanda GalambasEric M Shepard
Sep 8, 2019·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Amanda S ByerJoan B Broderick
Jul 6, 2020·The Journal of Biological Chemistry·Brigitta NémethGustav Berggren
Oct 14, 2018·International Journal of Molecular Sciences·Marco BortolusDonatella Carbonera
Jul 10, 2021·Dalton Transactions : an International Journal of Inorganic Chemistry·Eric M ShepardJoan B Broderick
Aug 31, 2017·Biochemistry·Eric M ShepardJoan B Broderick

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Datasets Mentioned

BETA
ESR935

Methods Mentioned

BETA
X-ray
GTPase
circular dichroism
size-exclusion chromatography
Gel filtration

Software Mentioned

HydE
SATMON
OriginPro Fit Peaks (
OriginPro Fit Peaks
MONMER
MENOSR
OriginPro

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