{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["8"],"submitter":["Winkler M"],"pubmed_abstract":["H<sub>2</sub> turnover at the [FeFe]-hydrogenase cofactor (H-cluster) is assumed to follow a reversible heterolytic mechanism, first yielding a proton and a hydrido-species which again is double-oxidized to release another proton. Three of the four presumed catalytic intermediates (H<sub>ox</sub>, H<sub>red</sub>/H<sub>red</sub> and H<sub>sred</sub>) were characterized, using various spectroscopic techniques. However, in catalytically active enzyme, the state containing the hydrido-species, which is eponymous for the proposed heterolytic mechanism, has yet only been speculated about. We use different strategies to trap and spectroscopically characterize this transient hydride state (H<sub>hyd</sub>) for three wild-type [FeFe]-hydrogenases. Applying a novel set-up for real-time attenuated total-reflection Fourier-transform infrared spectroscopy, we monitor compositional changes in the state-specific infrared signatures of [FeFe]-hydrogenases, varying buffer pH and gas composition. We selectively enrich the equilibrium concentration of H<sub>hyd</sub>, applying Le Chatelier's principle by simultaneously increasing substrate and product concentrations (H<sub>2</sub>/H<sup>+</sup>). Site-directed manipulation, targeting either the proton-transfer pathway or the adt ligand, significantly enhances H<sub>hyd</sub> accumulation independent of pH."],"journal":["Nature communications"],"pagination":["16115"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC5524980"],"repository":["biostudies-literature"],"pubmed_title":["Accumulating the hydride state in the catalytic cycle of [FeFe]-hydrogenases."],"pmcid":["PMC5524980"],"pubmed_authors":["Duan J","Hofmann E","Stripp ST","Senger M","Wittkamp F","Winkler M","Happe T","Apfel UP","Esselborn J"],"additional_accession":[]},"is_claimable":false,"name":"Accumulating the hydride state in the catalytic cycle of [FeFe]-hydrogenases.","description":"H<sub>2</sub> turnover at the [FeFe]-hydrogenase cofactor (H-cluster) is assumed to follow a reversible heterolytic mechanism, first yielding a proton and a hydrido-species which again is double-oxidized to release another proton. Three of the four presumed catalytic intermediates (H<sub>ox</sub>, H<sub>red</sub>/H<sub>red</sub> and H<sub>sred</sub>) were characterized, using various spectroscopic techniques. However, in catalytically active enzyme, the state containing the hydrido-species, which is eponymous for the proposed heterolytic mechanism, has yet only been speculated about. We use different strategies to trap and spectroscopically characterize this transient hydride state (H<sub>hyd</sub>) for three wild-type [FeFe]-hydrogenases. Applying a novel set-up for real-time attenuated total-reflection Fourier-transform infrared spectroscopy, we monitor compositional changes in the state-specific infrared signatures of [FeFe]-hydrogenases, varying buffer pH and gas composition. We selectively enrich the equilibrium concentration of H<sub>hyd</sub>, applying Le Chatelier's principle by simultaneously increasing substrate and product concentrations (H<sub>2</sub>/H<sup>+</sup>). Site-directed manipulation, targeting either the proton-transfer pathway or the adt ligand, significantly enhances H<sub>hyd</sub> accumulation independent of pH.","dates":{"release":"2017-01-01T00:00:00Z","publication":"2017 Jul","modification":"2024-11-05T17:35:26.466Z","creation":"2019-03-27T02:51:30Z"},"accession":"S-EPMC5524980","cross_references":{"pubmed":["28722011"],"doi":["10.1038/ncomms16115"]}}