Discovery of Dark pH-Dependent H+ Migration in a [NiFe]-Hydrogenase and Its Mechanistic Relevance: Mobilizing the Hydrido Ligand of the Ni-C Intermediate

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• 作者：Bonnie J. Murphy ; Ricardo Hidalgo ; Maxie M. Roessler ; Rhiannon M. Evans ; Philip A. Ash ; William K. Myers ; Kylie A. Vincent ; Fraser A. Armstrong
• 刊名：Journal of the American Chemical Society
• 出版年：2015
• 出版时间：July 8, 2015
• 年：2015
• 卷：137
• 期：26
• 页码：8484-8489
• 全文大小：383K
• ISSN：1520-5126

文摘

Despite extensive studies on [NiFe]-hydrogenases, the mechanism by which these enzymes produce and activate H₂ so efficiently remains unclear. A well-known EPR-active state produced under H₂ and known as Ni-C is assigned as a Ni^III鈥揊e^II species with a hydrido ligand in the bridging position between the two metals. It has long been known that low-temperature photolysis of Ni-C yields distinctive EPR-active states, collectively termed Ni-L, that are attributed to migration of the bridging-H species as a proton; however, Ni-L has mainly been regarded as an artifact with no mechanistic relevance. It is now demonstrated, based on EPR and infrared spectroscopic studies, that the Ni-C to Ni-L interconversion in Hydrogenase-1 (Hyd-1) from Escherichia coli is a pH-dependent process that proceeds readily in the dark鈥攑roton migration from Ni-C being favored as the pH is increased. The persistence of Ni-L in Hyd-1 must relate to unassigned differences in proton affinities of metal and adjacent amino acid sites, although the unusually high reduction potentials of the adjacent Fe鈥揝 centers in this O₂-tolerant hydrogenase might also be a contributory factor, impeding elementary electron transfer off the [NiFe] site after proton departure. The results provide compelling evidence that Ni-L is a true, albeit elusive, catalytic intermediate of [NiFe]-hydrogenases.