title: Proton-coupled electron transfer dynamics in the catalytic mechanism of a [NiFe]-hydrogenase.
abstract: The movement of protons and electrons is common to the synthesis of all chemical fuels such as H2. Hydrogenases, which catalyze the reversible reduction of protons, necessitate transport and reactivity between protons and electrons, but a detailed mechanism has thus far been elusive. Here, we use a phototriggered chemical potential jump method to rapidly initiate the proton reduction activity of a [NiFe] hydrogenase. Coupling the photochemical initiation approach to nanosecond transient infrared and visible absorbance spectroscopy afforded direct observation of interfacial electron transfer and active site chemistry. Tuning of intramolecular proton transport by pH and isotopic substitution revealed distinct concerted and stepwise proton-coupled electron transfer mechanisms in catalysis. The observed heterogeneity in the two sequential proton-associated reduction processes suggests a highly engineered protein environment modulating catalysis and implicates three new reaction intermediates; Nia-I, Nia-D, and Nia-SR(-). The results establish an elementary mechanistic understanding of catalysis in a [NiFe] hydrogenase with implications in enzymatic proton-coupled electron transfer and biomimetic catalyst design.
authors: †Chemistry Department, Emory University, Atlanta, Georgia 30322, United States.
authors: Brandon L
authors: BL
authors: Greene
authors: ‡Department of Biochemistry, University of Georgia, Athens, Georgia 30602, United States.
authors: Chang-Hao
authors: CH
authors: Wu
authors: ‡Department of Biochemistry, University of Georgia, Athens, Georgia 30602, United States.
authors: Patrick M
authors: PM
authors: McTernan
authors: ‡Department of Biochemistry, University of Georgia, Athens, Georgia 30602, United States.
authors: Michael W W
authors: MW
authors: Adams
authors: †Chemistry Department, Emory University, Atlanta, Georgia 30322, United States.
authors: R Brian
authors: RB
authors: Dyer
mesh_terms: D055162:Biocatalysis; D020134:Catalytic Domain; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D006864:Hydrogenase / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D055668:Photochemical Processes; D011522:Protons; D020027:Pyrococcus furiosus / Q000201:enzymology; D013696:Temperature
chemical_list: D011522:Protons; C043296:nickel-iron hydrogenase; D006864:Hydrogenase