8w6x
Neutron structure of [NiFe]-hydrogenase from D. vulgaris Miyazaki F in its oxidized state
Structural highlights
FunctionPHNL_NITV9 Catalyzes the reversible oxidoreduction of molecular hydrogen, in conjunction with a specific electron acceptor, cytochrome c3. Publication Abstract from PubMed[NiFe]-hydrogenase from Desulfovibrio vulgaris Miyazaki F is an O(2)-sensitive enzyme that is inactivated in the presence of O(2) but the oxidized enzyme can recover its catalytic activity by reacting with H(2) under anaerobic conditions. Here, we report the first neutron structure of [NiFe]-hydrogenase in its oxidized state, determined at a resolution of 2.20 A. This resolution allowed us to reinvestigate the structure of the oxidized active site and to observe the positions of protons in several short hydrogen bonds. X-ray anomalous scattering data revealed that a part of the Ni ion is dissociated from the active site Ni-Fe complex and forms a new square-planar Ni complex, accompanied by rearrangement of the coordinated thiolate ligands. One of the thiolate Sgamma atoms is oxidized to a sulfenate anion but remains attached to the Ni ion, which was evaluated by quantum chemical calculations. These results suggest that the square-planar complex can be generated by the attack of reactive oxygen species derived from O(2), as distinct from one-electron oxidation leading to a conventional oxidized form of the Ni-Fe complex. Another major finding of this neutron structure analysis is that the Cys17(S) thiolate Sgamma atom coordinating to the proximal Fe-S cluster forms an unusual hydrogen bond with the main-chain amide N atom of Gly19(S) with a distance of 3.25 A, where the amide proton appears to be delocalized between the donor and acceptor atoms. This observation provides insight into the contribution of the coordinated thiolate ligands to the redox reaction of the Fe-S cluster. New insights into the oxidation process from neutron and X-ray crystal structures of an O(2)-sensitive [NiFe]-hydrogenase.,Hiromoto T, Nishikawa K, Inoue S, Ogata H, Hori Y, Kusaka K, Hirano Y, Kurihara K, Shigeta Y, Tamada T, Higuchi Y Chem Sci. 2023 Aug 21;14(35):9306-9315. doi: 10.1039/d3sc02156d. eCollection 2023 , Sep 13. PMID:37712026[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
| ||||||||||||||||||||
