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Issue 9, 2010
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Comparison of the membrane-bound [NiFe] hydrogenases from R. eutropha H16 and D. vulgaris Miyazaki F in the oxidized ready state by pulsed EPR

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Abstract

The geometric and electronic structures of the active sites in the oxidized Nir–B state of the [NiFe] hydrogenases from Ralstonia eutropha H16 and Desulfovibrio vulgaris Miyazaki F were investigated in pulsed EPR and ENDOR experiments at two different microwave frequencies (X- and Q-band). Two hyperfine-couplings were clearly resolved in the frozen solution spectra arising from the β-protons of the nickel-coordinating cysteine residues Cys549 and Cys586 from the Desulfovibrio vulgaris and Ralstonia eutropha hydrogenase, respectively. ESEEM spectroscopic experiments reveal the presence of a histidine in the second coordination sphere of the Ni. The spectroscopic data indicate that the electronic structures of the [NiFe] centers in both hydrogenases are identical in the Nir–B state. However, additional spin couplings of the active site to further paramagnetic centers were identified for the Ralstonia eutropha hydrogenase. The respective couplings could be clearly resolved and simulated. The results from this study are discussed in view of the exceptional O2-tolerance of the Ralstonia hydrogenase.

Graphical abstract: Comparison of the membrane-bound [NiFe] hydrogenases from R. eutropha H16 and D. vulgaris Miyazaki F in the oxidized ready state by pulsed EPR

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Publication details

The article was received on 23 Oct 2009, accepted on 07 Dec 2009 and first published on 15 Jan 2010


Article type: Paper
DOI: 10.1039/B922236G
Citation: Phys. Chem. Chem. Phys., 2010,12, 2139-2148

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    Comparison of the membrane-bound [NiFe] hydrogenases from R. eutropha H16 and D. vulgaris Miyazaki F in the oxidized ready state by pulsed EPR

    M. Saggu, C. Teutloff, M. Ludwig, M. Brecht, M. Pandelia, O. Lenz, B. Friedrich, W. Lubitz, P. Hildebrandt, F. Lendzian and R. Bittl, Phys. Chem. Chem. Phys., 2010, 12, 2139
    DOI: 10.1039/B922236G

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