Deciphering catalytic mechanism of [FeFe]-hydrogenase by neutron crystallography
Final Report Abstract
[FeFe]-hydrogenases efficiently catalyze the reversible conversion of protons, electrons and dihydrogen (H2) employing their catalytic cofactor termed H-cluster. Catalytic turnover at the deeply buried H-cluster within protein matrix is achieved by concerted proton coupled electron transfer steps. To understand the fate of catalytically important hydrogen species binding to the H-cluster and the proton transfer pathway, neutron crystallography was proposed to be employed based on the well-established X-ray crystallography of in the hosting labs. Although the large-sized CpI ([FeFe]-hydrogenase from Clostridium pasteurianum) crystals (>0.1 mm3) were obtained, their diffraction capacity decreased to resolutions of 2.8-3.5 Å with high mosacity, indicating severe problems in crystal packing. Despite a lot of attempted efforts in optimization, large-sized CpI crystals suitable for neutron diffraction were not obtained in the end. More extensive screening for crystallization is needed in the future. In addition, the applicant has made other progress in structural studies on [FeFe]- hydrogenases. Extrinsic cyanide was demonstrated to directly bind at the open coordination site of the H-cluster. In the cyanide bound state, an alternative conformation of the proton transfer pathway was observed, possibly reflecting a second functional conformation of the proton transfer pathway. Formaldehyde was shown to react with the amine group of the H-cluster and the thiol group of the proton transfer pathway which both are catalytically very important for catalytic turnover, therefore providing a molecular mechanism about how formaldehyde inhibits [FeFe]-hydrogenases. A structural and functional understanding of the oxygen-stable [FeFe]-hydrogenase from Clostridium beijerinckii was obtained.
Publications
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Electrochemical control of [FeFe]-hydrogenase single crystals reveals complex redox populations at the catalytic site. Dalton Transactions, 50(36), 12655-12663.
Morra, Simone; Duan, Jifu; Winkler, Martin; Ash, Philip A.; Happe, Thomas & Vincent, Kylie A.
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Following Electroenzymatic Hydrogen Production by Rotating Ring–Disk Electrochemistry and Mass Spectrometry**. Angewandte Chemie International Edition, 60(18), 10001-10006.
Khushvakov, Jaloliddin; Nussbaum, Robin; Cadoux, Cécile; Duan, Jifu; Stripp, Sven T. & Milton, Ross D.
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Trapping an Oxidized and Protonated Intermediate of the [FeFe]-Hydrogenase Cofactor under Mildly Reducing Conditions. Inorganic Chemistry, 61(26), 10036-10042.
Senger, Moritz; Duan, Jifu; Pavliuk, Mariia V.; Apfel, Ulf-Peter; Haumann, Michael & Stripp, Sven T.
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A Dynamic Water Channel Affects O2 Stability in [FeFe]‐Hydrogenases. ChemSusChem, 17(3).
Brocks, Claudia; Das, Chandan K.; Duan, Jifu; Yadav, Shanika; Apfel, Ulf‐Peter; Ghosh, Subhasri; Hofmann, Eckhard; Winkler, Martin; Engelbrecht, Vera; Schäfer, Lars V. & Happe, Thomas
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Cyanide Binding to [FeFe]‐Hydrogenase Stabilizes the Alternative Configuration of the Proton Transfer Pathway. Angewandte Chemie International Edition, 62(7).
Duan, Jifu; Hemschemeier, Anja; Burr, David J.; Stripp, Sven T.; Hofmann, Eckhard & Happe, Thomas
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Insights into the Molecular Mechanism of Formaldehyde Inhibition of [FeFe]-Hydrogenases. Journal of the American Chemical Society, 145(48), 26068-26074.
Duan, Jifu; Veliju, Astrit; Lampret, Oliver; Liu, Lingling; Yadav, Shanika; Apfel, Ulf-Peter; Armstrong, Fraser A.; Hemschemeier, Anja & Hofmann, Eckhard
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Structural determinants of oxygen resistance and Zn 2+ -mediated stability of the [FeFe]-hydrogenase from Clostridium beijerinckii. Proceedings of the National Academy of Sciences, 122(3).
Duan, Jifu; Rutz, Andreas; Kawamoto, Akihiro; Naskar, Shuvankar; Edenharter, Kristina; Leimkühler, Silke; Hofmann, Eckhard; Happe, Thomas & Kurisu, Genji
