Characterization of the Hydrogen-Deuterium Exchange Activities of the Energy-Transducing HupSL Hydrogenase and H
            <sub>2</sub>
            -Signaling HupUV Hydrogenase in
            <i>Rhodobacter capsulatus</i>

Vignais, Paulette M.; Dimon, Bernard; Зорин, Н. А.; Tomiyama, Masamitsu; Colbeau, Annette

doi:10.1128/jb.182.21.5997-6004.2000

Cited by 44 publications

(52 citation statements)

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“…Its synthesis is activated under both aerobiosis and anaerobiosis by the presence of its substrate H 2 , either produced endogenously by nitrogenase (as a by-product) or added exogenously (12). The regulatory cascade responding to H 2 has been thoroughly studied in R. capsulatus (13,16,18,47) and Ralstonia eutropha (3,23,26,27), and comprises three proteins: HupUV, HupT, and HupR (HoxBC, HoxJ, and HoxA, respectively, in R. eutropha). In R. capsulatus, these proteins are encoded by four genes located on the chromosome in the locus hup.…”

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confidence: 99%

“…In particular, all of the residues able to bind the Fe-S clusters in HupS and the bimetallic center in the HupL active site of Desulfovibrio gigas [NiFe]hydrogenase (48) are conserved in HupUV. Recent data showed that this protein was able to catalyze the three typical reactions of the hydrogenases: namely H 2 uptake and H 2 evolution in presence of electron acceptor or donor, respectively, and hydrogen-deuterium exchange in their absence (46,47). HupUV is a soluble protein and does not contain the N-terminal signal peptide, which addresses the hydrogenases to or across the membrane.…”

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confidence: 99%

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Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

2003

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The photosynthetic bacterium Rhodobacter capsulatus contains two [NiFe]hydrogenases: an energy-generating hydrogenase, HupSL, and a regulatory hydrogenase, HupUV. The synthesis of HupSL is specifically activated by H 2 through a signal transduction cascade comprising three proteins: the H 2 -sensing HupUV protein, the histidine kinase HupT, and the transcriptional regulator HupR. Whereas a phosphotransfer between HupT and HupR was previously demonstrated, interaction between HupUV and HupT was only hypothesized based on in vivo analyses of mutant phenotypes. To visualize the in vitro interaction between HupUV and HupT proteins, a six-His (His 6 )-HupU fusion protein and the HupV protein were coproduced by using a homologous expression system. The two proteins copurified as a His 6 -HupUHupV complex present in dimeric and tetrameric forms, both of which had H 2 uptake activity. We demonstrated that HupT and HupUV interact and form stable complexes that could be separated on a native gel. Interaction was also monitored with surface plasmon resonance technology and was shown to be insensitive to salt concentration and pH changes, suggesting that the interactions involve hydrophobic residues. As expected, H 2 affects the interaction between HupUV and HupT, leading to a weakening of the interaction, which is independent of the phosphate status of HupT. Several forms of HupT were tested for their ability to interact with HupUV and to complement hupT mutants. Strong interaction with HupUV was obtained with the isolated PAS domain of HupT and with inactive HupT mutated in the phosphorylable histidine residue, but only the wild-type HupT protein was able to restore normal H 2 regulation.

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Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

2003

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“…The synthesis of R. capsulatus HupUV requires at least HypF and HypD (Vignais et al, 2000), and that of R. eutropha HoxBC is highly dependent on HypDEF . These proteins (and also Ech from M. barkeri, Coo from R. rubrum and Coo from C. hydrogenoformans) lack the carboxyl-terminal extension cleaved in the precursor form of [NiFe]-H 2 ase large subunits.…”

Section: Metallocenter Assemblymentioning

confidence: 99%

“…Two representatives of group 2, the Rhodobacter capsulatus HupUV H 2 ase (Vignais et al, 2000) and the R. eutropha HoxBC H 2 ase (Kleihues et al, 2000) are indeed soluble, cytoplasmic proteins. They are not directly involved in energy transducing reactions.…”

Section: The Cytoplasmic H 2 Sensors and The Cyanobacterial Uptake [Nmentioning

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Molecular Biology of Microbial Hydrogenases

2004

Current Issues in Molecular Biology

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Catalytic Bias in Enzymatic Metal Cofactor‐Based Oxidation–Reduction Catalysis

Varghese¹,

Mulder²,

Raugei³

et al. 2021

Encyclopedia of Inorganic and Bioinorganic Chemistry

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Catalytic bias refers to the relative rate preference of a catalyst for either the forward or reverse direction. In enzymatic metal cofactor‐based oxidation–reduction catalysis, the tuning of catalytic bias plays an underlying role in controlling rates of reactivity. For this, enzymes have evolved complex active sites that can exist in multiple oxidation states with differing reduction potentials in order to achieve challenging multi‐step, oxidation–reduction reactions. Conceivably, the relative stability of the intermediates that contribute to determining the rate‐limiting step of the catalytic cycle could impose catalytic bias, although mechanisms for this concept are just beginning to be realized. As one example, recent work on Clostridium pasteurianum [FeFe]‐hydrogenases which catalyze reversible hydrogen oxidation have shown that the differential stabilization/destabilization of active site oxidation states through either static or dynamic protein interactions can preferentially promote either the hydrogen oxidation or proton reduction direction of the reaction. This revealed how an enzymatic cofactor can impose bias in oxidation–reduction catalysis through various tuning mechanisms by protein scaffold interactions. The hypothesis based on achieving catalytic bias through the modulation of cofactor oxidation states critical for the reaction cycle can be extended more generally to other cofactor‐based oxidation–reductions catalysts. The current understanding of catalytic bias has significant implications for the design of synthetic catalysts used in industrial settings, as well as providing a greater fundamental understanding of the factors that control metabolic processes in all life.

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Characterization of the Hydrogen-Deuterium Exchange Activities of the Energy-Transducing HupSL Hydrogenase and H ₂ -Signaling HupUV Hydrogenase in Rhodobacter capsulatus

Cited by 44 publications

References 40 publications

Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

Molecular Biology of Microbial Hydrogenases

Catalytic Bias in Enzymatic Metal Cofactor‐Based Oxidation–Reduction Catalysis

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Characterization of the Hydrogen-Deuterium Exchange Activities of the Energy-Transducing HupSL Hydrogenase and H 2 -Signaling HupUV Hydrogenase in Rhodobacter capsulatus

Cited by 44 publications

References 40 publications

Interaction between the H 2 Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

Interaction between the H 2 Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

Molecular Biology of Microbial Hydrogenases

Catalytic Bias in Enzymatic Metal Cofactor‐Based Oxidation–Reduction Catalysis

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Characterization of the Hydrogen-Deuterium Exchange Activities of the Energy-Transducing HupSL Hydrogenase and H ₂ -Signaling HupUV Hydrogenase in Rhodobacter capsulatus

Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus

Interaction between the H ₂ Sensor HupUV and the Histidine Kinase HupT Controls HupSL HydrogenaseSynthesis in Rhodobactercapsulatus