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Main Title: Hydroxy-bridged resting states of a [NiFe]-hydrogenase unraveled by cryogenic vibrational spectroscopy and DFT computations
Author(s): Caserta, Giorgio
Pelmenschikov, Vladimir
Lorent, Christian
Waffo, Armel F. Tadjoung
Katz, Sagie
Lauterbach, Lars
Schoknecht, Janna
Wang, Hongxin
Yoda, Yoshitaka
Tamasaku, Kenji
Kaupp, Martin
Hildebrandt, Peter
Lenz, Oliver
Cramer, Stephen P.
Zebger, Ingo
Type: Article
Language Code: en
Abstract: The catalytic mechanism of [NiFe]-hydrogenases is a subject of extensive research. Apart from at least four reaction intermediates of H2/H+ cycling, there are also a number of resting states, which are formed under oxidizing conditions. Although not directly involved in the catalytic cycle, the knowledge of their molecular structures and reactivity is important, because these states usually accumulate in the course of hydrogenase purification and may also play a role in vivo during hydrogenase maturation. Here, we applied low-temperature infrared (cryo-IR) and nuclear resonance vibrational spectroscopy (NRVS) to the isolated catalytic subunit (HoxC) of the heterodimeric regulatory [NiFe]-hydrogenase (RH) from Ralstonia eutropha. Cryo-IR spectroscopy revealed that the HoxC protein can be enriched in almost pure resting redox states suitable for NRVS investigation. NRVS analysis of the hydrogenase catalytic center is usually hampered by strong spectral contributions of the FeS clusters of the small, electron-transferring subunit. Therefore, our approach to investigate the FeS cluster-free, 57Fe-labeled HoxC provided an unprecedented insight into the [NiFe] site modes, revealing their contributions in a spectral range otherwise superimposed by FeS cluster-derived bands. Rationalized by density functional theory (DFT) calculations, our data provide structural descriptions of the previously uncharacterized hydroxy- and water-containing resting states. Our work highlights the relevance of cryogenic vibrational spectroscopy and DFT to elucidate the structure of barely defined redox states of the [NiFe]-hydrogenase active site.
Issue Date: 11-Dec-2020
Date Available: 10-Jun-2021
DDC Class: 540 Chemie und zugeordnete Wissenschaften
Subject(s): [NiFe]-hydrogenase
DFT computations
Sponsor/Funder: DFG, 390540038, EXC 2008: Unifying Systems in Catalysis "UniSysCat"
EC/H2020/810856/EU/Twin to Illuminate Metals in Biology and Biocatalysis through Biospectroscopy/TIMB3
Journal Title: Chemical Science
Publisher: Royal Society of Chemistry
Publisher Place: Cambridge
Volume: 12
Issue: 6
Publisher DOI: 10.1039/D0SC05022A
Page Start: 2189
Page End: 2197
EISSN: 2041-6539
ISSN: 2041-6520
Appears in Collections:FG Physikalische Chemie / Biophysikalische Chemie » Publications

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