Overview of the Maturation Machinery of the H-Cluster of [FeFe]-Hydrogenases with a Focus on HydF

Bortolus, Marco; Costantini, Paola; Doni, Davide; Carbonera, Donatella

doi:10.3390/ijms19103118

Cited by 20 publications

(14 citation statements)

References 75 publications

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“…5 and Fig. S6 ; see also reference 37 ). To investigate the possibility for HydA1 to interact with the tetrameric form of HydF, a comparison was made between the docked HydA1-HydF complex and the published crystal structure of tetrameric T. neapolitana HydF (PDB entry 3QQ5 ) ( 22 ).…”

Section: Resultsmentioning

confidence: 89%

The maturase HydF enables [FeFe] hydrogenase assembly via transient, cofactor-dependent interactions

Németh

Land

Magnuson

et al. 2020

Journal of Biological Chemistry

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[FeFe] hydrogenases have attracted extensive attention in the field of renewable energy research due to their remarkable efficiency for H2 gas production. H2 formation is catalyzed by a biologically unique hexanuclear iron cofactor denoted the “H-cluster”. The assembly of this cofactor requires a dedicated maturation machinery including HydF, a multidomain [4Fe4S] cluster protein with GTPase activity. HydF is responsible for harboring and delivering a pre-catalyst to the apo-hydrogenase, but the details of this process are not well understood. Herein we utilize Gas-phase Electrophoretic MacroMolecule Analysis to show that a HydF dimer forms a transient interaction complex with the hydrogenase, and that the formation of this complex is dependent on the cofactor content on HydF. Moreover, FTIR, EPR and UV/Vis spectroscopy studies of mutants of HydF show that the isolated iron-sulfur cluster domain retains the capacity for binding the pre-catalyst in a reversible fashion, and is capable of activating apo-hydrogenase in in vitro assays. These results demonstrate the central role of the iron-sulfur cluster domain of HydF in final stages of H-cluster assembly, i.e. in binding and delivering the pre-catalyst.

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Section: Resultsmentioning

confidence: 89%

The maturase HydF enables [FeFe] hydrogenase assembly via transient, cofactor-dependent interactions

Németh

Land

Magnuson

et al. 2020

Journal of Biological Chemistry

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“…HydG synthesizes the 2 low-spin, mononuclear iron synthons ([Fe(CO) 2 CN]) as subunits of the [2Fe H ] precursor (21). At the end of the HydG (αβ) 8 triosephosphate isomerase (TIM) barrel, a cubane [4Fe-4S] cluster binds SAM (22,23). At the opposite end of the active site cavity, another auxiliary FeS-site can be observed, being either a [4Fe-5S] or a [5Fe-5S] cluster, the latter one corresponds to a cubane cluster that is coupled via a μ 2 -sulfide to a mononuclear Fe II -center.…”

mentioning

confidence: 99%

The final steps of [FeFe]-hydrogenase maturation

Lampret

Esselborn

Haas

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The active site (H-cluster) of [FeFe]-hydrogenases is a blueprint for the design of a biologically inspired H2-producing catalyst. The maturation process describes the preassembly and uptake of the unique [2FeH] cluster into apo-hydrogenase, which is to date not fully understood. In this study, we targeted individual amino acids by site-directed mutagenesis in the [FeFe]-hydrogenase CpI of Clostridium pasteurianum to reveal the final steps of H-cluster maturation occurring within apo-hydrogenase. We identified putative key positions for cofactor uptake and the subsequent structural reorganization that stabilizes the [2FeH] cofactor in its functional coordination sphere. Our results suggest that functional integration of the negatively charged [2FeH] precursor requires the positive charges and individual structural features of the 2 basic residues of arginine 449 and lysine 358, which mark the entrance and terminus of the maturation channel, respectively. The results obtained for 5 glycine-to-histidine exchange variants within a flexible loop region provide compelling evidence that the glycine residues function as hinge positions in the refolding process, which closes the secondary ligand sphere of the [2FeH] cofactor and the maturation channel. The conserved structural motifs investigated here shed light on the interplay between the secondary ligand sphere and catalytic cofactor.

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“…Correspondingly, it has been confirmed that the purified HydF, together with HydE and HydG, was able to convert apo‐HydA into an active [FeFe]‐hydrogenase . The maturases “HydE, HydF, and HydG” are able to synthesize and attach to the H‐cluster . However, only HydE and HydG are the members of radical S‐adenosyl methionine (SAM) family, and both contains a [4Fe‐4S] cluster near the N‐terminus which serve as an open binding site of a SAM cofactor.…”

Section: Hydrogen Synthesizing Enzymes In Microalgaementioning

confidence: 97%

“…In addition, HydE and HydG both contain another iron sulfur cluster as C‐terminus binding motif, which can be occupied by a [2Fe‐2S] cluster in HydE but a [4Fe‐4S] cluster in HydG. Differently, HydF is composed of a [4Fe‐4S] cluster near the C‐terminus cysteines and a N‐terminal GTPase domain for hydrogenase maturation . Although the minute maturation mechanism is still not completely understood, an appealing assumption is that HydE and HydG synthesize the ligands of the binuclear subsite, while HydF may be a scaffold for the assembly of the binuclear subsite.…”

Section: Hydrogen Synthesizing Enzymes In Microalgaementioning

confidence: 99%

Microalgal Hydrogen Production

et al. 2020

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Microalgae, as facultative aerobic organisms, convert solar energy to produce biohydrogen under anaerobic condition. Biohydrogen is a kind of ideal clean and renewable energy source with great commercial potential. Many endeavors have been focused on improving the biohydrogen yields by various means. Here, the research history of hydrogen production by microalgae (including cyanobacteria and green algae), the characteristics of nitrogenase and hydrogenase, the mechanisms of hydrogen production, the technological progress, and the application of enzymatic, genetic, and metabolic engineering methods to improve hydrogen production by microalgae are reviewed. In addition, the regulation of anaerobic metabolisms of Chlamydomonas reinhardtii after the disruption of key enzymes functioning in the fermentative pathways is discussed. Finally, the main challenges and obstacles facing the more efficient production and commercialization of hydrogen production from microalgae in the future are proposed.

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Overview of the Maturation Machinery of the H-Cluster of [FeFe]-Hydrogenases with a Focus on HydF

Cited by 20 publications

References 75 publications

The maturase HydF enables [FeFe] hydrogenase assembly via transient, cofactor-dependent interactions

The maturase HydF enables [FeFe] hydrogenase assembly via transient, cofactor-dependent interactions

The final steps of [FeFe]-hydrogenase maturation

Microalgal Hydrogen Production

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