Demethylmenaquinol is a substrate of Escherichia coli nitrate reductase A (NarGHI) and forms a stable semiquinone intermediate at the NarGHI quinol oxidation site

Rendon, Julia; Pilet, Eric; Fahs, Zeinab; Seduk, Farida; Sylvi, Léa; Chehade, Mahmoud Hajj; Pierrel, Fabien; Guigliarelli, Bruno; Magalon, Axel; Grimaldi, Stéphane

doi:10.1016/j.bbabio.2015.05.001

Cited by 10 publications

(24 citation statements)

References 55 publications

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“…[41,74] Figure S4. ChemPhysChem 2017ChemPhysChem , 18,2704ChemPhysChem -2714 www.chemphyschem.org For NarGHI overproduction, the strains were transformed with the pVA700 plasmid and grown semi-aerobically at 37 8Ci nd efined minimal medium [51] supplemented by either [methyl-2 H] methionine (35 mm,i sotope purity 98 %, Cambridge Isotope Laboratories, Inc), l-lysine-e-15 N·2HCl (136 mm,i sotope purity 98 %, euriso-top), or l-histidine-d-15 N·HCl·H 2 O( 142 mm;i sotope purity 98 %, eurisotop) and by 0.2 mm of isopropyl 1-thio-b-d-galactopyranoside to induce the narGHJI expression. Typically,c ells were first grown aerobically at 37 8Cuntil stationary phase in 60 mL LB medium supplemented with casaminoacids (0.05 %) and containing kanamycin (30 mgmL À1 ), spectinomycin (25 mgmL À1 ), and ampicillin (100 mgmL À1 ).…”

Section: Experimental Section Bacterialstrains Plasmids and Growth mentioning

confidence: 99%

“…By combining site directedm utagenesis experiments, redox potentiometry,a nd EPR techniques, we have demonstrated that E. coli NarGHI stabilizes in the Q D site the semiquinone form of the three respiratory quinones availablei nt his organism, namely menasemiquinone (MSK),u bisemiquinone (USQ), and demethylmenasemiquinone (DMSK). [18,46,50,51] Moreover, their radical intermediates exhibit as imilar interaction with a single 14 Nn ucleus in their 14 NH YSCORE spectra. The nuclear quadrupolep arameters (k, h)w ere accurately determined by 3GHz ESEEM/HYSCORE experiments for the MSK at the Q D site (hereafter referred to as MSK D ), leadingt o( k, h) = (0.49 MHz, 0.5).…”

Section: Introductionmentioning

confidence: 99%

“…[18,50,51] It exhibitst wo cross peaks (1 in Figure 2A)c orrelating nuclear transition frequencies at 2.2 and 3.4 MHz from opposite electron spin manifolds. These were assigned to the two double-quantum transition frequencies from aw eakly coupled 14 Nn ucleusw ith A iso ( 14 N) % 0.8 MHz.…”

Section: 1mentioning

confidence: 99%

“…[18,46,50,51] Moreover, their radical intermediates exhibit as imilar interaction with a single 14 Nn ucleus in their 14 NH YSCORE spectra. The nuclear quadrupolep arameters (k, h)w ere accurately determined by 3GHz ESEEM/HYSCORE experiments for the MSK at the Q D site (hereafter referred to as MSK D ), leadingt o( k, h) = (0.49 MHz, 0.5).…”

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confidence: 99%

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confidence: 99%

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Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

et al. 2017

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In vivo specific isotope labeling at the residue or substituent level is used to probe menasemiquinone (MSK) binding to the quinol oxidation site of respiratory nitrate reductase A (NarGHI) from E. coli. 15N selective labeling of His15Nδ or Lys15Nζ in combination with hyperfine sublevel correlation (HYSCORE) spectroscopy unambiguously identified His15Nδ as the direct hydrogen‐bond donor to the radical. In contrast, an essentially anisotropic coupling to Lys15Nζ consistent with a through‐space magnetic interaction was resolved. This suggests that MSK does not form a hydrogen bond with the side chain of the nearby Lys86 residue. In addition, selective 2H labeling of the menaquinone methyl ring substituent allows unambiguous characterization of the 2H—and hence of the 1H—methyl isotropic hyperfine coupling by 2H HYSCORE. DFT calculations show that a simple molecular model consisting of an imidazole Nδ atom in a hydrogen‐bond interaction with a MSK radical anion satisfactorily accounts for the available spectroscopic data. These results support our previously proposed one‐sided binding model for MSK to NarGHI through a single short hydrogen bond to the Nδ of His66, one of the distal heme axial ligands. This work establishes the basis for future investigations aimed at determining the functional relevance of this peculiar binding mode.

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Section: Experimental Section Bacterialstrains Plasmids and Growth mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: 1mentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

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Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate‐Based Inhibitors of Cytochrome bo₃ Ubiquinol Oxidase from Escherichia coli

et al. 2020

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The respiratory chain of Escherichia coli contains two different types of terminal oxidase that are differentially regulated as a response to changing environmental conditions. These oxidoreductases catalyze the reduction of molecular oxygen to water and contribute to the proton motive force. The cytochrome bo3 oxidase (cyt bo3) acts as the primary terminal oxidase under atmospheric oxygen levels, whereas the bd‐type oxidase is most abundant under microaerobic conditions. In E. coli, both types of respiratory terminal oxidase (HCO and bd‐type) use ubiquinol‐8 as electron donor. Here, we assess the inhibitory potential of newly designed and synthesized 3‐alkylated Lawson derivatives through L‐proline‐catalyzed three‐component reductive alkylation (TCRA). The inhibitory effects of these Lawson derivatives on the terminal oxidases of E. coli (cyt bo3 and cyt bd‐I) were tested potentiometrically. Four compounds were able to reduce the oxidoreductase activity of cyt bo3 by more than 50 % without affecting the cyt bd‐I activity. Moreover, two inhibitors for both cyt bo3 and cyt bd‐I oxidase could be identified. Based on molecular‐docking simulations, we propose binding modes of the new Lawson inhibitors. The molecular fragment benzyl enhances the inhibitory potential and selectivity for cyt bo3, whereas heterocycles reduce this effect. This work extends the library of 3‐alkylated Lawson derivatives as selective inhibitors for respiratory oxidases and provides molecular probes for detailed investigations of the mechanisms of respiratory‐chain enzymes of E. coli.

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Direct Electrochemistry of Molybdenum and Tungsten Enzymes

Fourmond

2018

Encyclopedia of Interfacial Chemistry

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International audienceMolybdenum and tungsten enzymes are almost omnipresent in living organisms; they catalyze a large variety of reactions on a large range of substrates, and are involved in a number of key cellular functions, like bacterial respiration, carbon, sulfur, and nitrogen cycles, and detoxification. Most of these enzymes catalyze redox reactions, and they have successfully been connected to electrodes, in order to not only build biotechnological devices like biosensors or biofuel cells but also to learn about their catalytic properties, using protein film voltammetry, in which the electron transfer is direct and the enzymatic activity can be monitored as an electrical current. This chapter is an exhaustive review of the uses of direct electrochemistry for the study of molybdenum and tungsten enzymes

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Demethylmenaquinol is a substrate of Escherichia coli nitrate reductase A (NarGHI) and forms a stable semiquinone intermediate at the NarGHI quinol oxidation site

Cited by 10 publications

References 55 publications

Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate‐Based Inhibitors of Cytochrome bo₃ Ubiquinol Oxidase from Escherichia coli

Direct Electrochemistry of Molybdenum and Tungsten Enzymes

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Demethylmenaquinol is a substrate of Escherichia coli nitrate reductase A (NarGHI) and forms a stable semiquinone intermediate at the NarGHI quinol oxidation site

Cited by 10 publications

References 55 publications

Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective 2H and 15N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective 2H and 15N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate‐Based Inhibitors of Cytochrome bo3 Ubiquinol Oxidase from Escherichia coli

Direct Electrochemistry of Molybdenum and Tungsten Enzymes

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Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Probing the Menasemiquinone Binding Mode to Nitrate Reductase A by Selective ²H and ¹⁵N Labeling, HYSCORE Spectroscopy, and DFT Modeling

Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate‐Based Inhibitors of Cytochrome bo₃ Ubiquinol Oxidase from Escherichia coli