A single-crystal ENDOR and density functional theory study of the oxidized states of the [NiFe] hydrogenase from Desulfovibrio vulgaris Miyazaki F |
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Authors: | Maurice van Gastel Matthias Stein Marc Brecht Olga Schröder Friedhelm Lendzian Robert Bittl Hideaki Ogata Yoshiki Higuchi Wolfgang Lubitz |
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Institution: | 1. Max-Planck-Institut für Bioanorganische Chemie, Stiftstrasse 34-36, 45470, Mülheim an der Ruhr, Germany 2. Max-Volmer-Laboratorium für Biophysikalische Chemie, Fakult?t für Mathematik und Naturwissenschaften, Technische Universit?t Berlin, 10623, Berlin, Germany 3. EML Research gGmbH, Villa Bosch, Schloss-Wolfsbrunnenweg 33, 69118, Heidelberg, Germany 4. Institut für Experimentalphysik, Fachbereich Physik, Freie Universit?t Berlin, Arnimallee 14, 14195, Berlin, Germany 5. University of Hyogo and Himeji Institute of Technology, 3-2-1 Kouto, Kamigori-cho, Ako-gun, Hyogo, 678-1297, Japan
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Abstract: | The catalytic center of the NiFe] hydrogenase of Desulfovibrio vulgaris Miyazaki F in the oxidized states was investigated by electron paramagnetic resonance and electron–nuclear double resonance
spectroscopy applied to single crystals of the enzyme. The experimental results were compared with density functional theory
(DFT) calculations. For the Ni-B state, three hyperfine tensors could be determined. Two tensors have large isotropic hyperfine
coupling constants and are assigned to the β-CH2 protons of the Cys-549 that provides one of the bridging sulfur ligands between Ni and Fe in the active center. From a comparison
of the orientation of the third hyperfine tensor with the tensor obtained from DFT calculations an OH− bridging ligand has been identified in the Ni-B state. For the Ni-A state broader signals were observed. The signals of the
third proton, as observed for the “ready” state Ni-B, were not observed at the same spectral position for Ni-A, confirming
a structural difference involving the bridging ligand in the “unready” state of the enzyme.
Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.
Maurice van Gastel and Matthias Stein contributed equally to this work. |
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Keywords: | Electron– nuclear double resonance Hydrogenase Single crystal |
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