Changing the Ligation of the Distal [4Fe4S] Cluster in NiFe Hydrogenase Impairs Inter- and Intramolecular Electron Transfers

Dementin, Sébastien; Belle, Valérie; Bertrand, Patrick; Guigliarelli, Bruno; Adryanczyk-Perrier, Géraldine; Lacey, Antonio L. De; Fernández, Vı́ctor M.; Rousset, Marc; Léger, Christophe

doi:10.1021/ja060233b

Cited by 99 publications

(164 citation statements)

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“…Distances from distal iron-sulfur cluster to graphite surface (Z), overlap integrals of donor and acceptor frontier orbitals (S), electronic coupling matrices (VDA), and calculated rates of ET (kET). 4 15.3 2.9 x 10 -6 1.1 x 10 -2 3.6 x 10 2 > 40 Electrochemical investigation of the WT and an all-cysteine coordinated hydrogenase mutant gave significantly reduced rates of electron transfer from the distal [FeS]d-cluster to the electrode [18]. Our calculated value of 360 s -1 is in excellent agreement with the experimental the results of > 40 s -1 .…”

Section: Distal Fes-clustersupporting

confidence: 73%

Distal [FeS]-Cluster Coordination in [NiFe]-Hydrogenase Facilitates Intermolecular Electron Transfer

Petrenko¹,

Stein²

2017

Preprint

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Biohydrogen is a versatile energy carrier for the generation of electric energy from renewable sources. Hydrogenases can be used in enzymatic fuel cells to oxidize dihydrogen. The rate of electron transfer (ET) at the anodic side between the [NiFe]-hydrogenase enzyme distal ironsulfur cluster and the electrode surface can be described by the Marcus equation. All parameters for the Marcus equation are accessible from DFT calculations. The distal cubane FeS-cluster has a three cysteine and one histidine coordination [Fe4S4](His)(Cys)3 first ligation sphere. The reorganization energy (inner-and outer-sphere) is almost unchanged upon a histidine-to-cysteine substitution. Differences in rates of electron transfer between the wild-type enzyme and the all-cysteine mutant can be rationalized by a diminished electronic coupling between the donor and acceptor molecules in the [Fe4S4](Cys)4 case. The fast and efficient electron transfer from the distal iron-sulfur cluster is realized by a fine-tuned protein environment which facilitates the flow of electrons. This study enables the design and control of electron transfer rates and pathways by protein engineering.

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Section: Distal Fes-clustersupporting

confidence: 73%

Distal [FeS]-Cluster Coordination in [NiFe]-Hydrogenase Facilitates Intermolecular Electron Transfer

Petrenko¹,

Stein²

2017

Preprint

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“…In 2006, Léger and co-workers [59] investigated mutations to the ligation of the distal [4Fe-4S] cluster of hydrogenase, which is presumably the first port-of-call for ET to or from the enzyme at an electrode surface (figure 6a). The authors interestingly note that the k 0 of the enzyme is modified upon replacing the single His ligation of the distal cluster with a Gly, where a diminished direct bioelectrocatalytic signal is observed (figure 6b).…”

Section: Direct Bioelectrocatalysis Of Metalloenzymesmentioning

confidence: 99%

Direct enzymatic bioelectrocatalysis: differentiating between myth and reality

Milton

Minteer

2017

J. R. Soc. Interface.

158

148

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Enzymatic bioelectrocatalysis is being increasingly exploited to better understand oxidoreductase enzymes, to develop minimalistic yet specific biosensor platforms, and to develop alternative energy conversion devices and bioelectrosynthetic devices for the production of energy and/or important chemical commodities. In some cases, these enzymes are able to electronically communicate with an appropriately designed electrode surface without the requirement of an electron mediator to shuttle electrons between the enzyme and electrode. This phenomenon has been termed direct electron transfer or direct bioelectrocatalysis. While many thorough studies have extensively investigated this fascinating feat, it is sometimes difficult to differentiate desirable enzymatic bioelectrocatalysis from electrocatalysis deriving from inactivated enzyme that may have also released its catalytic cofactor. This article will review direct bioelectrocatalysis of several oxidoreductases, with an emphasis on experiments that provide support for direct bioelectrocatalysis versus denatured enzyme or dissociated cofactor. Finally, this review will conclude with a series of proposed control experiments that could be adopted to discern successful direct electronic communication of an enzyme from its denatured counterpart.

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“…In fact, when a H106A mutant (which corresponds to H101A in this study) in the P. denitrificans NuoG homologue (Nqo3) was overexpressed, almost no iron-sulfur clusters were incorporated into the subunit, making further investigations at the subunit level impossible (13). There is another report that the His to Cys variant of Clostridium acetobutylicum iron-only hydrogenase, which has the same binding motif as cluster N5, was not produced in quantities large enough for structural and thermodynamic studies (36). In this respect, it was fortunate for us that the Fe/S clusters remained intact (Fig.…”

Section: The Mutation Of Fe/s Clusters Often Causes Destabilization-mentioning

confidence: 85%

“…In the periplasmic NiFe hydrogenases, when the histidine (His-184 in Desulfovibrio fructosovorans) in the binding motif "HXXCX 24 CXXXXXC" was changed to a cysteine or glycine, the iron-sulfur cluster was still assembled, but the electron transfer activity of the mutants was almost completely lost (36). For the H184G variant, the activity was partially restored by adding exogenous ligands such as imidazole (36).…”

Section: The Mutation Of Fe/s Clusters Often Causes Destabilization-mentioning

confidence: 99%

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Iron-Sulfur Cluster N5 Is Coordinated by an HXXXCXXCXXXXXC Motif in the NuoG Subunit of Escherichia coli NADH:Quinone Oxidoreductase (Complex I)

Nakamaru‐Ogiso

Matsuno‐Yagi

Yagi

et al. 2008

Journal of Biological Chemistry

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, and N6b were misassigned. Here we identified and characterized cluster N5 in the Escherichia coli complex I whose EPR signals had never been detected by any group. Using homologous recombination, we constructed mutant strains of H101A, H101C, H101A/C114A, and cluster N5 knock-out. Although mutant NuoEFG subcomplexes were dissociated from complex I, we successfully recovered these mutant NuoCDEFG subcomplexes by expressing the His-tagged NuoCD subunit, which had a high affinity to NuoG. The W221A mutant was used as a control subcomplex carrying wild-type clusters. By lowering temperatures to around 3 K, we finally succeeded in detecting cluster N5 signals in the control for the first time. However, no cluster N5 signals were found in any of the N5 mutants, whereas EPR signals from all other clusters were detected. These data confirmed that, contrary to the misassignment claim, cluster N5 has a unique coordination with His(Cys) 3 ligands in NuoG.The proton-translocating NADH:ubiquinone oxidoreductase (EC 1.6.5.3) (complex I) is the largest energy-transducing complex in the aerobic respiratory chains of many prokaryotes and eukaryotes (1-3). Complex I is one of the most complicated and elaborate iron-sulfur (Fe/S)3 proteins yet known (4). Recently, the three-dimensional structure of the hydrophilic domain of Thermus thermophilus HB-8 complex I has been determined at 3.3 Å resolution (5). It revealed the spatial localization of all of the redox centers and their coordinating amino acid residues. For the sake of simplicity, we will use the Escherichia coli nomenclature for each subunit. The primary electron acceptor of complex I is a noncovalently bound flavin mononucleotide (FMN (Fig. 1A) (5, 6). These seven Fe/S clusters are lined up to form the main electron transfer pathway, whereas a [2Fe-2S] cluster N1a in NuoE and a [4Fe-4S] cluster N7 in NuoG stick out from the main pathway. It has been suggested that clusters N1a and N7 may have different roles (5, 7). The sequence analysis has confirmed that all of the Fe/S clusters except cluster N7 are conserved (8).EPR spectroscopy has been most informative for the analysis of Fe/S clusters. However, because the sensitivity and resolution of EPR spectroscopy are much lower than those of spectrophotometry, considerable spectral overlaps exist. Therefore, to make a definitive assignment of the spectra to each of the specific Fe/S clusters is sometimes very difficult. In addition, some Fe/S clusters may not be detectable by EPR when the spin relaxation time is too short, or when the Fe/S cluster is not paramagnetic under certain chemical or electronic conditions. In fact, E. coli complex I contains at least six EPR-detectable Fe/S clusters as follows: N1a, N1b, N2, N3, N4, and N7. However, N5 signals have not been detected so far. Cluster N6a and N6b signals have not been characterized sufficiently. Another problem is that EPR identification of an Fe/S cluster residing in the overexpressed single subunit could be misleading, because its EPR signals may be altered from tho...

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Changing the Ligation of the Distal [4Fe4S] Cluster in NiFe Hydrogenase Impairs Inter- and Intramolecular Electron Transfers

Cited by 99 publications

References 53 publications

Distal [FeS]-Cluster Coordination in [NiFe]-Hydrogenase Facilitates Intermolecular Electron Transfer

Distal [FeS]-Cluster Coordination in [NiFe]-Hydrogenase Facilitates Intermolecular Electron Transfer

Direct enzymatic bioelectrocatalysis: differentiating between myth and reality

Iron-Sulfur Cluster N5 Is Coordinated by an HXXXCXXCXXXXXC Motif in the NuoG Subunit of Escherichia coli NADH:Quinone Oxidoreductase (Complex I)

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