Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O-2-tolerant NAD(+)-reducing [NiFe] hydrogenase

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dc.contributor.authorLauterbach, Lars
dc.contributor.authorWang, Hongxin
dc.contributor.authorHorch, Marius
dc.contributor.authorGee, Leland B.
dc.contributor.authorYoda, Yoshitaka
dc.contributor.authorTanaka, Yoshihito
dc.contributor.authorZebger, Ingo
dc.contributor.authorLenz, Oliver
dc.contributor.authorCramer, Stephen P.
dc.date.accessioned2017-10-25T06:29:13Z
dc.date.available2017-10-25T06:29:13Z
dc.date.issued2015
dc.description.abstractHydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD(+)-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H-2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by Fe-57 specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, which is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD(+)-reducing hydrogenases. For the first time, Fe-CO and Fe-CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective C-13 labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe-CO modes. The present approach explores the complex vibrational signature of the Fe-S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.en
dc.description.sponsorshipDFG, EXC 314, Unifying Concepts in Catalysisen
dc.identifier.eissn2041-6539
dc.identifier.issn2041-6520
dc.identifier.pmid25678951
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/6978
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-6317
dc.language.isoen
dc.rights.urihttps://creativecommons.org/licenses/by-nc/3.0/
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftende
dc.titleNuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O-2-tolerant NAD(+)-reducing [NiFe] hydrogenaseen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.doi10.1039/c4sc02982h
dcterms.bibliographicCitation.issue2
dcterms.bibliographicCitation.journaltitleChemical Scienceen
dcterms.bibliographicCitation.originalpublishernameRoyal Society of Chemistryde
dcterms.bibliographicCitation.originalpublisherplaceCambridgede
dcterms.bibliographicCitation.pageend1060
dcterms.bibliographicCitation.pagestart1055
dcterms.bibliographicCitation.volume6
tub.accessrights.dnbfree
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 Berlin

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