Influence of the [4Fe–4S] cluster coordinating cysteines on active site maturation and catalytic properties of <i>C. reinhardtii</i> [FeFe]-hydrogenase

Kertess, Leonie; Adamska-Venkatesh, Agnieszka; Rodríguez‐Maciá, Patricia; Rüdiger, Olaf; Lubitz, Wolfgang; Happe, Thomas

doi:10.1039/c7sc03444j

Cited by 27 publications

(46 citation statements)

References 57 publications

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“…Enzyme Activity Assays. Hydrogen-production activity assays were performed as described previously (54). A 400-ng sample of holo-hydrogenase was incubated in 2 mL of reaction mixture, containing KpI buffer (0.1 M potassium phosphate, pH 6.8), NaDT (0.1 M), and methyl viologen (10 mM) using sealed 8-mL vessels (Suba).…”

Section: Preparation Of [2fe H ] Mimics and In Vitro Maturation Of [Fmentioning

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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Section: Preparation Of [2fe H ] Mimics and In Vitro Maturation Of [Fmentioning

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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“…5 In this paper " bidirectional " implies that the catalysts can catalyse the reaction in both directions ( figure 1 B and C), as opposed to only one-way ( figure 1 A). We call "catalytic preference" 4 the ratio of the maximal currents measured at high driving force in both directions (in the enzyme electrochemistry field, and particularly regarding hydrogenases, the expression "catalytic bias" has been used [6][7][8][9][10][11][12][13][14][15] ). Since these two rates are measured under different experimental conditions, their ratio is not the equilibrium constant of the reaction; it is not set by the thermodynamics of the reaction, but depends on the catalyst's properties instead.…”

Section: Introductionmentioning

confidence: 99%

Understanding and Design of Bidirectional and Reversible Catalysts of Multielectron, Multistep Reactions

et al. 2019

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HAL is a multidisciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L'archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Distributed under a Creative Commons Attribution-NonCommercial-NoDerivatives| 4.0 International License

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“…Beyond this, all these studies opened the way for the study of direct catalysis using enzymes at electrochemical interfaces. 30 , 31 …”

Section: Introductionmentioning

confidence: 99%

Electron transfer in an acidophilic bacterium: interaction between a diheme cytochrome and a cupredoxin

et al. 2018

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Influence of the [4Fe–4S] cluster coordinating cysteines on active site maturation and catalytic properties of C. reinhardtii [FeFe]-hydrogenase

Abstract: Alteration of the [4Fe–4S] cluster coordinating cysteines reveals their individual importance for [4Fe–4S] cluster binding, [2Fe] insertion and catalytic turnover.

Cited by 27 publications

References 57 publications

The final steps of [FeFe]-hydrogenase maturation

The final steps of [FeFe]-hydrogenase maturation

Understanding and Design of Bidirectional and Reversible Catalysts of Multielectron, Multistep Reactions

Electron transfer in an acidophilic bacterium: interaction between a diheme cytochrome and a cupredoxin

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