Hydroxy-bridged resting states of a [NiFe]-hydrogenase unraveled by cryogenic vibrational spectroscopy and DFT computations

dc.contributor.authorCaserta, Giorgio
dc.contributor.authorPelmenschikov, Vladimir
dc.contributor.authorLorent, Christian
dc.contributor.authorWaffo, Armel F. Tadjoung
dc.contributor.authorKatz, Sagie
dc.contributor.authorLauterbach, Lars
dc.contributor.authorSchoknecht, Janna
dc.contributor.authorWang, Hongxin
dc.contributor.authorYoda, Yoshitaka
dc.contributor.authorTamasaku, Kenji
dc.contributor.authorKaupp, Martin
dc.contributor.authorHildebrandt, Peter
dc.contributor.authorLenz, Oliver
dc.contributor.authorCramer, Stephen P.
dc.contributor.authorZebger, Ingo
dc.date.accessioned2021-06-10T07:10:18Z
dc.date.available2021-06-10T07:10:18Z
dc.date.issued2020-12-11
dc.description.abstractThe 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.en
dc.description.sponsorshipDFG, 390540038, EXC 2008: Unifying Systems in Catalysis "UniSysCat"en
dc.description.sponsorshipEC/H2020/810856/EU/Twin to Illuminate Metals in Biology and Biocatalysis through Biospectroscopy/TIMB3en
dc.identifier.eissn2041-6539
dc.identifier.issn2041-6520
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/13229
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-12024
dc.language.isoenen
dc.rights.urihttps://creativecommons.org/licenses/by-nc/3.0/en
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftende
dc.subject.other[NiFe]-hydrogenaseen
dc.subject.otherspectroscopyen
dc.subject.otherDFT computationsen
dc.titleHydroxy-bridged resting states of a [NiFe]-hydrogenase unraveled by cryogenic vibrational spectroscopy and DFT computationsen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.doi10.1039/D0SC05022Aen
dcterms.bibliographicCitation.issue6en
dcterms.bibliographicCitation.journaltitleChemical Scienceen
dcterms.bibliographicCitation.originalpublishernameRoyal Society of Chemistryen
dcterms.bibliographicCitation.originalpublisherplaceCambridgeen
dcterms.bibliographicCitation.pageend2197en
dcterms.bibliographicCitation.pagestart2189en
dcterms.bibliographicCitation.volume12en
tub.accessrights.dnbfreeen
tub.affiliationFak. 2 Mathematik und Naturwissenschaften::Inst. Chemie::FG Physikalische Chemie / Biophysikalische Chemiede
tub.affiliation.facultyFak. 2 Mathematik und Naturwissenschaftende
tub.affiliation.groupFG Physikalische Chemie / Biophysikalische Chemiede
tub.affiliation.instituteInst. Chemiede
tub.publisher.universityorinstitutionTechnische Universität Berlinen

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