| Author |
Lenz, Oliver Parkin, Alison Armstrong, Fraser A. Sigfridsson, Kajsa G. V. Rousset, Marc Nishihara, Hirofumi De Lacey, Antonio L. Haumann, Michael Yoon, Ki-seok Chernev, Petko Leidel, Nils Sanganas, Oliver Dementin, Sébastien |
| Description |
Author Affiliation: Sigfridsson KG ( Freie Universität Berlin, Institut für Experimentalphysik, 14195 Berlin, Germany.); Leidel N ( Freie Universität Berlin, Institut für Experimentalphysik, 14195 Berlin, Germany.); Sanganas O ( Freie Universität Berlin, Institut für Experimentalphysik, 14195 Berlin, Germany.); Chernev P ( Freie Universität Berlin, Institut für Experimentalphysik, 14195 Berlin, Germany.); Lenz O ( Humboldt-Universität zu Berlin, Mikrobiologie, 10115 Berlin, Germany.); Yoon KS ( Ibaraki University, Department of Bioresource Science, Ibaraki 300-0393, Japan.); Nishihara H ( Ibaraki University, Department of Bioresource Science, Ibaraki 300-0393, Japan.); Parkin A ( University of Oxford, Department of Inorganic Chemistry, Oxford OX1 3QR, UK.); Armstrong FA ( University of Oxford, Department of Inorganic Chemistry, Oxford OX1 3QR, UK.); Dementin S ( CNRS Marseille, Unité de Bioénergétique et Ingénierie des Protéines, 13402 Marseille, France.); Rousset M ( CNRS Marseille, Unité de Bioénergétique et Ingénierie des Protéines, 13402 Marseille, France.); De Lacey AL ( Instituto de Catálisis (CSIC), 28049 Madrid, Spain.); Haumann M ( Freie Universität Berlin, Institut für Experimentalphysik, 14195 Berlin, Germany. Electronic address: michael.haumann@fu-berlin.de.) |
| Abstract |
The class of [NiFe]-hydrogenases comprises oxygen-sensitive periplasmic (PH) and oxygen-tolerant membrane-bound (MBH) enzymes. For three PHs and four MBHs from six bacterial species, structural features of the nickel–iron active site of hydrogen turnover and of the iron–sulfur clusters functioning in electron transfer were determined using X-ray absorption spectroscopy (XAS). Fe-XAS indicated surplus oxidized iron and a lower number of ~ 2.7 Å Fe–Fe distances plus additional shorter and longer distances in the oxidized MBHs compared to the oxidized PHs. This supported a double-oxidized and modified proximal FeS cluster in all MBHs with an apparent trimer-plus-monomer arrangement of its four iron atoms, in agreement with crystal data showing a [4Fe3S] cluster instead of a [4Fe4S] cubane as in the PHs. Ni-XAS indicated coordination of the nickel by the thiol group sulfurs of four conserved cysteines and at least one iron–oxygen bond in both MBH and PH proteins. Structural differences of the oxidized inactive [NiFe] cofactor of MBHs in the Ni-B state compared to PHs in the Ni–A state included a ~ 0.05 Å longer Ni-O bond, a two times larger spread of the Ni–S bond lengths, and a ~ 0.1 Å shorter Ni–Fe distance. The modified proximal [4Fe3S] cluster, weaker binding of the Ni–Fe bridging oxygen species, and an altered localization of reduced oxygen species at the active site may each contribute to O $_{2}$ tolerance. |
