Initial Electron Transfer Step Research Articles

Mechanisms for Methane and Ammonia Oxidation by Particulate Methane Monooxygenase.

Particulate MMO (pMMO) catalyzes the oxidation of methane to methanol and also ammonia to hydroxylamine. Experimental characterization of the active site has been very difficult partly because the enzyme is membrane-bound. However, recently, there has been major progress mainly through the use of cryogenic electron microscopy (cryoEM). Electron paramagnetic resonance (EPR) and X-ray spectroscopy have also been employed. Surprisingly, the active site has only one copper. There are two histidine ligands and one asparagine ligand, and the active site is surrounded by phenyl alanines but no charged amino acids in the close surrounding. The present study is the first quantum chemical study using a model of that active site (CuD). Low barrier mechanisms have been found, where an important part is that there are two initial proton-coupled electron transfer steps to a bound O2 ligand before the substrate enters. Surprisingly, this leads to large radical character for the oxygens even though they are protonated. That result is very important for the ability to accept a proton from the substrates. Methods have been used which have been thoroughly tested for redox enzyme mechanisms.

Bismuthene with stable Bi[sbnd]O bonds for efficient CO2 electroreduction to formate

Atomic configuration engineering is a promising strategy to realize the efficient and precise CO2 electroreduction towards the targeted products. Herein, an in-situ electrochemical transformation strategy was performed to construct the atomically thin bismuthene nanosheets with rich and stabilized BiO bonds (OBi-ene) from Bi2O2S precursor for CO2 electroreduction to formate. Electrochemical and thermodynamic characterizations confirm that rich BiO bonds could greatly enhance the CO2 adsorption on catalyst surface, which significantly accelerates the initial electron-transfer step of CO2 electroreduction for forming the key CO2·- intermediate. Consequently, the OBi-ene catalyst exhibits both high Faraday efficiency (>90%) and cathodic energy efficiency (>60%) to produce formate at wide current range from 50 to 400 mA cm−2 in flow cell. Impressively, a large current density of 200 mA cm−2 can be achieved only at 2.98 V in a full cell. This work provides a feasible strategy to design Bi-based electrocatalysts for efficient CO2 electroreduction.

Mechanistic dichotomies in redox reactions of mononuclear metal-oxygen intermediates.

There are mechanistic dichotomies with regard to the formation, electronic structures and reaction mechanisms of metal-oxygen intermediates, since these metal-oxygen species could be composed of different resonance structures or canonical structures of the oxidation states of metals and ligands, which may undergo different reaction pathways. Even the same metal-oxygen intermediates, such as metal-oxo species, may undergo an electron-transfer pathway or a direct hydrogen or oxygen atom transfer pathway depending on the one-electron redox potentials of metal-oxo species and substrates. Electron-transfer pathways are also classified into two mechanisms, such as outer-sphere and inner-sphere pathways. The one-electron redox potentials of metal-oxygen species and substrates are also shifted because of the binding of acids, which can result from either hydrogen bonding or protonation. There are a rebound pathway and a non-rebound pathway following the initial electron transfer or hydrogen atom transfer step to produce hydroxylated products, depending on the one-electron redox potentials of metal-oxo species and substrates. Nucleophilic reactions can be switched to electrophilic pathways, depending on reaction conditions such as reaction temperature. Spin states of metal-oxygen intermediates are also an important factor that controls the redox reactivity of oxidants in oxidation reactions. Here, we review such various mechanistic dichotomies in redox reactions of metal-oxygen intermediates with the emphasis on understanding and controlling the redox reactivity of metal-oxygen intermediates from experimental and theoretical points of view.

Ligand Noninnocence in Nickel Porphyrins: Nickel Isobacteriochlorin Formation under Hydrogen Evolution Conditions.

An electron-deficient nickel porphyrin complex undergoes facile ring reduction to form a nickel isobacteriochlorin complex under hydrogen evolution conditions. Spectroscopic experiments indicate that the reduced nickel porphyrin undergoes subsequent reduction and protonation to form a phlorin anion rather than a metal hydride, demonstrating that the key initial proton-coupled electron transfer step is directed toward the ligand versus the metal. The phlorin anion facilely converts to the isobacteriochlorin in the presence of two-electron and three-proton equivalents. Cyclic voltammetry (CV) and spectroscopic experiments reveal that the four-electron, four-proton electrochemical reduction of nickel porphyrin to isobacteriochlorin occurs promptly in the presence of the strong proton donor tosic acid, followed by hydrogen evolution reaction (HER) catalysis at slightly more negative potentials. CVs of independently synthesized Ni isobacteriochlorin show a catalytic HER at the same potentials as those observed for the HER in CVs of the Ni porphyrin. We find that, under strongly acidic conditions, the HER catalysis arises from conversion of the Ni isobacteriochlorin into a nickel-containing, catalytically active electrode-adsorbed species. These results show that Ni porphyrin converts to Ni isobacteriochlorin under HER catalysis conditions via a ligand-based PCET process and that it is the isobacteriochlorin complex which gives rise to an active HER catalysis.

Concepts and tools for mechanism and selectivity analysis in synthetic organic electrochemistry

As an accompaniment to the current renaissance of synthetic organic electrochemistry, the heterogeneous and space-dependent nature of electrochemical reactions is analyzed in detail. The reactions that follow the initial electron transfer step and yield the products are intimately coupled with reactant transport. Depiction of the ensuing reactions profiles is the key to the mechanism and selectivity parameters. Analysis is eased by the steady state resulting from coupling of diffusion with convection forced by solution stirring or circulation. Homogeneous molecular catalysis of organic electrochemical reactions of the redox or chemical type may be treated in the same manner. The same benchmarking procedures recently developed for the activation of small molecules in the context of modern energy challenges lead to the establishment and comparison of the catalytic Tafel plots. At the very opposite, redox-neutral chemical reactions may be catalyzed by injection (or removal) of an electron from the electrode. This class of reactions has currently few, but very thoroughly analyzed, examples. It is likely that new cases will emerge in the near future.

Advances in bipolar electrochemiluminescence for the detection of biorelevant molecular targets

Abstract Bipolar electrochemistry (BPE) contrasts very much with conventional electrochemistry because it is based on the control of the solution potential instead of the working electrode potential. In a typical setup, a piece of conducting materials is immersed in an electrolyte and submitted to an electric field. Such conditions split the interfacial nature of the materials into cathodic and anodic domains where electrochemical reactions can readily take place. BPE has many potential applications, and the present contribution aims to focus on recent analytical applications that involve electrogenerated chemiluminescence (ECL) detection. ECL is a special case of luminescence where the excited state of the luminophore is populated after a sequence of reaction that is triggered by an initial electron transfer step occurring at the electrode surface. The coupling between BPE and ECL is a powerful approach because it provides a unique opportunity to combine the intrinsic advantages of both techniques. BPE enables the spatial separation of sensing and reporting poles, whereas ECL provides a simple and sensitive visual readout. This opinion article will describe the experimental possibilities and the most recent applications of BPE/ECL coupling for the detection of biorelevant molecular targets.

Local and Global Electric Field Asymmetry in Photosynthetic Reaction Centers.

The origin of unidirectional electron transfer in photosynthetic reaction centers (RCs) has been widely discussed. Despite the high level of structural similarity between the two branches of pigments that participate in the initial electron transfer steps of photosynthesis, electron transfer only occurs along one branch. One possible explanation for this functional asymmetry is the differences in the electrostatic environment between the active and the inactive branches arising from the charges and dipoles of the organized protein structure. We present an analysis of electric fields in the RC of the purple bacterium Rhodobacter sphaeroides using the intrinsic carbonyl groups of the pigments as vibrational reporters whose vibrational frequency shifts can be converted into electric fields based on the vibrational Stark effect and also provide Stark effect data for plant pigments that can be used in future studies. The carbonyl stretches of the isolated pigments show pronounced Stark effects. We use these data, solvatochromism, molecular dynamics simulations, and data in the literature from IR and Raman spectra to evaluate differences in fields at symmetry-related positions, in particular at the 9-keto and 2-acetyl positions of the pigments involved in primary charge separation.

Enzymatic Systems with Homology to Nitrogenase: Biosynthesis of Bacteriochlorophyll and Coenzyme F430.

Enzymes with homology to nitrogenase are essential for the reduction of chemically stable double bonds within the biosynthetic pathways of bacteriochlorophyll and coenzyme F430. These tetrapyrrole-based compounds are crucial for bacterial photosynthesis and the biogenesis of methane in methanogenic archaea. Formation of bacteriochlorophyll requires the unique ATP-dependent enzyme chlorophyllide oxidoreductase (COR) for the two-electron reduction of chlorophyllide to bacteriochlorophyllide. COR catalysis is based on the homodimeric protein subunit BchX2, which facilitates the transfer of electrons to the corresponding heterotetrameric catalytic subunit (BchY/BchZ)2. By analogy to the nitrogenase system, the dynamic switch protein BchX2 contains a [4Fe-4S] cluster that triggers the ATP-driven transfer of electrons onto a second [4Fe-4S] cluster located in (BchY/BchZ)2. The subsequent substrate reduction and protonation is unrelated to nitrogenase catalysis, with no further involvement of a molybdenum-containing cofactor. The biosynthesis of the nickel-containing coenzyme F430 includes the six-electron reduction of the tetrapyrrole macrocycle of Ni2+-sirohydrochlorin a,c-diamide to Ni2+-hexahydrosirohydrochlorin a,c-diamide catalyzed by CfbC/D. The homodimeric CfbC2 subunit carrying a [4Fe-4S] cluster shows close homology to BchX2. Accordingly, parallelism for the initial ATP-driven electron transfer steps of CfbC/D was proposed. Electrons are received by the dimeric catalytic subunit CfbD2, which contains a second [4Fe-4S] cluster and carries out the saturation of an overall of three double bonds in a highly orchestrated spatial and regioselective process. Following a short introduction to nitrogenase catalysis, this chapter will focus on the recent progress toward the understanding of the nitrogenase-like enzymes COR and CfbC/D, with special emphasis on the underlying enzymatic mechanism(s).

Oxidation of inorganic compounds by aqueous permanganate: Kinetics and initial electron transfer steps

Abstract In this study, the kinetics of the reactions between aqueous permanganate (Mn(VII)) and selected inorganic compounds (e.g. KSCN, NaNO2, Na2SO3, Na2S2O3, KI, N2H4, H2O2, and NH2OH) were investigated in aqueous solutions at pH 5–9 with excess pyrophosphate (L) by detecting the decay of Mn(VII) using a stopped-flow technique. With excess L, soluble Mn(III)L rather than MnO2 was generated, which had negligible interference on examining the change of the absorbance of Mn(VII) at 525 nm (∼45 M−1 cm−1 for Mn(III)L vs 2500 M−1 cm−1 for Mn(VII)). The reactions of Mn(VII) with Na2SO3, Na2S2O3, KSCN, NaNO2, and KI (referred to as SA) exhibited second-order kinetics. Comparatively, the reactions of Mn(VII) with N2H4, H2O2, and NH2OH (referred to as SB) showed two-phase kinetics (i.e., an initial lag phase and a secondary fast phase), and the second-order rate constants (k1, M−1 s−1) were calculated in the initial phase. In the second phase, Mn(III)L reacted with SB producing Mn(II), and then Mn(II) reacted fast with Mn(VII) producing Mn(III)L. The kinetics of the reaction between Mn(II) and Mn(VII) with excess L at pH 4–9 were examined and the loss rate of Mn(VII) increased with increasing pH value. The species-specific second-order rate constants (k1i, M−1 s−1) for the reactions of Mn(VII) with inorganic compounds were determined by the experimental k1 values. Furthermore, a linear free energy relationship (LFER) was found (log(k1i) = 2.86 − 1.90E(2)0) between log k1i and 2−e− reduction potentials for inorganic species (i.e., SO32−, S2O32−, SCN−, H2O2, NH2OH, I−, and N2H4) while only NO2− showed 1−e− transfer process, indicating highly active aqueous manganese intermediates (Mn(V) and Mn(VI)) formed in situ upon Mn(VII) reduction.

Visible Light Mediated Photoredox Catalytic Arylation Reactions.

Introducing aryl- and heteroaryl moieties into molecular scaffolds are often key steps in the syntheses of natural products, drugs, or functional materials. A variety of cross-coupling methods have been well established, mainly using transition metal mediated reactions between prefunctionalized substrates and arenes or C-H arylations with functionalization in only one coupling partner. Although highly developed, one drawback of the established sp2-sp2 arylations is the required transition metal catalyst, often in combination with specific ligands and additives. Therefore, photoredox mediated arylation methods have been developed as alternative over the past decade. We begin our survey with visible light photo-Meerwein arylation reactions, which allow C-H arylation of heteroarenes, enones, alkenes, and alkynes with organic dyes, such as eosin Y, as the photocatalyst. A good number of examples from different groups illustrate the broad application of the reaction in synthetic transformations. While initially only photo-Meerwein arylation-elimination processes were reported, the reaction was later extended to photo-Meerwein arylation-addition reactions giving access to the photoinduced three component synthesis of amides and esters from alkenes, aryl diazonium salts, nitriles or formamides, respectively. Other substrates with redox-active leaving groups have been explored in photocatalyzed arylation reactions, such as diaryliodonium and triarylsulfonium salts, and arylsulfonyl chlorides. We discus some examples with their scope and limitations. The scope of arylation reagents for photoredox reactions was extended to aryl halides. The challenge here is the extremely negative reduction potential of aryl halides in the initial electron transfer step compared to, e.g., aryl diazonium or diaryliodonium salts. In order to reach reduction potentials over -2.0 V vs SCE two consecutive photoinduced electron transfer steps were used. The intermediary formed colored radical anion of the organic dye perylenediimide is excited by a second photon allowing the one electron reduction of acceptor substituted aryl chlorides. The radical anion of the aryl halide fragments under the loss of a halide ion and the aryl radical undergoes C-H arylation with biologically important pyrrole derivatives or adds to a double bond. Rhodamine 6G as an organic photocatalyst allows an even higher degree of control of the reaction. The dye is photoreduced in the presence of an amine donor under irradiation with green light (e.g., 530 nm), yielding its radical anion, which is a mild reducing reagent. The hypsochromic shift of the absorption of the rhodamine 6G radical anion toward blue region of the visible light spectrum allows its selective excitation using blue light (e.g., 455 nm). The excited radical anion is highly reducing and able to activate even bromoanisole for C-H arylation reactions, although only in moderate yield. Photoredox catalytic C-H arylation reactions are valuable alternatives to metal catalyzed reactions. They have an excellent functional group tolerance, could potentially avoid metal containing catalysts, and use visible light as a traceless reagent for the activation of arylating reagents.

Breaking Bonds with Electrons and Protons. Models and Examples

Besides its theoretical interest, the attention currently aroused by proton-coupled electron transfers (PCET reactions) has two main motives. One is a better understanding of biological processes in which PCET reactions are involved, Photosystem II as well as a myriad of other natural systems. The other is directed toward synthetic processes, many of which are related to global energy challenges. Until recently, the analyses of the mechanism and reactivity of PCET reactions have focused on outersphere transfers, those in which no bond between heavy atoms (all atoms with the exception of H) is concomitantly formed or broken. Conversely, reactions in which electron transfer triggers the breaking of a heavy-atom bond with no proton transfer have been extensively analyzed, both theoretically and experimentally. In both cases, strategies have been developed to distinguish between stepwise and concerted pathways. In each case, kinetic models have been devised, allowing the relation between activation and thermodynamic driving force to be established by means of parameters pertaining to the initial and final state. Although many natural and artificial processes include electron transfer, proton transfer, and heavy-atom bond breaking (/formation), no means were offered until recently to analyze the mechanism of such reactions, notably to establish the degree of concertedness of the three constitutive events. Likewise, no kinetic models were available to describe reactions where the three events are concerted. In this Account, we discuss the strategies to distinguish stepwise, partially concerted (when two of the three events are concerted), and totally concerted pathways in these reactions that include electron transfer, proton transfer, and heavy-atom bond breaking. These mechanism analysis methods are illustrated and validated by three examples. First we describe the electrochemical cleavage of an O-O bond in an aliphatic peroxide molecule with a pendant carboxylic acid group that can serve as proton donor for electron transfer and bond breaking. In the second example, we examine the breaking of one of the C-O bonds of CO2 within a multistep process where the reduction of CO2 into CO is catalyzed by an electrogenerated iron(0) porphyrin in the presence of various Brönsted acids. In this case, an intramolecular electron transfer triggers proton transfer and bond cleavage. In the first two examples, all three events are concerted. The third example also involves catalysis. It describes the cleavage of a cobalt-carbon bond in the reduction of chloroacetonitrile catalyzed by an electrogenerated cobalt(I) porphyrin. It illustrates the rather common case where the intermediate formed by the reaction of a transition metal complex with the substrate has to be cleaved to close the catalytic cycle. In the first two examples, all three events are concerted, whereas, in the last case, a partially concerted pathway takes place: proton transfer and bond-breaking (Co-C cleavage) are concerted after an initial electron transfer step. The all-concerted cases require a model that connects the kinetics to the thermodynamic driving force of the reaction. Starting from previous models of outersphere electron transfer, concerted proton-electron transfer, and concerted dissociative electron transfer, we describe a model for all-concerted proton-electron-bond breaking reactions. These pathways skip the high-energy intermediates that occur in stepwise pathways, but could introduce kinetic penalties. The all-concerted model allows one to assess these penalties and the way in which they can be fought by the supplement of driving force offered by concerted proton transfer.

Ion imaging study of dissociative charge transfer in the N2+ + CH4 system

The velocity map ion imaging method is applied to the dissociative charge transfer reactions of N2(+) with CH4 studied in crossed beams. The velocity space images are collected at four collision energies between 0.5 and 1.5 eV, providing both product kinetic energy and angular distributions for the reaction products CH3(+) and CH2(+). The general shapes of the images are consistent with long range electron transfer from CH4 to N2(+) preceding dissociation, and product kinetic energy distributions are consistent with energy resonance in the initial electron transfer step. The branching ratio for CH3(+):CH2(+) is 85:15 over the full collision energy range, consistent with literature reports.

Structural Characterization of HP1264 Reveals a Novel Fold for the Flavin Mononucleotide Binding Protein

Complex I (NADH-quinone oxidoreductase) is an enzyme that catalyzes the initial electron transfer from nicotinamide adenine dinucleotide (NADH) to flavin mononucleotide (FMN) bound at the tip of the hydrophilic domain of complex I. The electron flow into complex I is coupled to the generation of a proton gradient across the membrane that is essential for the synthesis of ATP. However, Helicobacter pylori has an unusual complex I that lacks typical NQO1 and NQO2 subunits, both of which are generally included in the NADH dehydrogenase domain of complex I. Here, we determined the solution structure of HP1264, one of the unusual subunits of complex I from H. pylori, which is located in place of NQO2, by three-dimensional nuclear magnetic resonance (NMR) spectroscopy and revealed that HP1264 can bind to FMN through UV-visible, fluorescence, and NMR titration experiments. This result suggests that FMN-bound HP1264 could be involved in the initial electron transfer step of complex I. In addition, HP1264 is structurally most similar to Escherichia coli TusA, which belongs to the SirA-like superfamily having an IF3-like fold in the SCOP database, implying that HP1264 adopts a novel fold for FMN binding. On the basis of the NMR titration data, we propose the candidate residues Ile32, Met34, Leu58, Trp68, and Val71 of HP1264 for the interaction with FMN. Notably, these residues are not conserved in the FMN binding site of any other flavoproteins with known structure. This study of the relationship between the structure and FMN binding property of HP1264 will contribute to improving our understanding of flavoprotein structure and the electron transfer mechanism of complex I.

Toward Full Peptide Sequence Coverage by Dual Fragmentation Combining Electron-Transfer and Higher-Energy Collision Dissociation Tandem Mass Spectrometry

Increasing peptide sequence coverage by tandem mass spectrometry improves confidence in database search-based peptide identification and facilitates mapping of post-translational modifications and de novo sequencing. Inducing 2-fold fragmentation by combining electron-transfer and higher-energy collision dissociation (EThcD) generates dual fragment ion series and facilitates extensive peptide backbone fragmentation. After an initial electron-transfer dissociation step, all ions including the unreacted precursor ions are subjected to collision induced dissociation which yields b/y- and c/z-type fragment ions in a single spectrum. This new fragmentation scheme provides richer spectra and substantially increases the peptide sequence coverage and confidence in peptide identification.

Initial electron transfer steps in complex I

Initial electron transfer steps in complex I

Organized media effect on the photochemical deoxygenation of resazurin in the presence of triethanolamine

The photodeoxygenation of the synthetic dye resazurin in the presence of triethanolamine was investigated in water, CTAC and SDS direct micelles, AOT and BHDC reverse micelles, and soybean lecithin microemulsions (LEC). In all cases the only product observed was the deoxygenated dye resorufin. Triplet and reaction quantum yields were determined in all media. The photoreaction proceeds more efficiently in the microheterogenous systems with positive interface, CTAC and BHDC, while the lower yield is observed in AOT, SDS and LEC. The initial step in the mechanism is the interaction of the triplet state of the dye with the amine, and the effect of the interface is interpreted by a decrease of the recombination rate of the radicals formed in the initial electron transfer step. Negative and zwitterionic interfaces have no effect on the quantum yield.

Green Rust Reduction of Chromium Part 2: Comparison of Heterogeneous and Homogeneous Chromate Reduction

White and green rusts are the active chemical reagents of buried scrap iron pollutant remediation. In this work, a comparison of the initial electron-transfer step for the reduction of CrO4−2 by Fe...

Independent initiation of primary electron transfer in the two branches of the photosystem I reaction center

Photosystem I (PSI) is a large pigment-protein complex that unites a reaction center (RC) at the core with approximately 100 core antenna chlorophylls surrounding it. The RC is composed of two cofactor branches related by a pseudo-C2 symmetry axis. The ultimate electron donor, P(700) (a pair of chlorophylls), and the tertiary acceptor, F(X) (a Fe(4)S(4) cluster), are both located on this axis, while each of the two branches is made up of a pair of chlorophylls (ec2 and ec3) and a phylloquinone (PhQ). Based on the observed biphasic reduction of F(X), it has been suggested that both branches in PSI are competent for electron transfer (ET), but the nature and rate of the initial electron transfer steps have not been established. We report an ultrafast transient absorption study of Chlamydomonas reinhardtii mutants in which specific amino acids donating H-bonds to the 13(1)-keto oxygen of either ec3(A) (PsaA-Tyr696) or ec3(B) (PsaB-Tyr676) are converted to Phe, thus breaking the H-bond to a specific ec3 cofactor. We find that the rate of primary charge separation (CS) is lowered in both mutants, providing direct evidence that the primary ET event can be initiated independently in each branch. Furthermore, the data provide further support for the previously published model in which the initial CS event occurs within an ec2/ec3 pair, generating a primary ec2(+)ec3(-) radical pair, followed by rapid reduction by P(700) in the second ET step. A unique kinetic modeling approach allows estimation of the individual ET rates within the two cofactor branches.

Fluorophenyl-substituted Fe-only hydrogenases active site ADT models: different electrocatalytic process for proton reduction in HOAc and HBF4/Et2O

A set of fluorophenyl-substituted adt-bridged Fe2S2 active site models of Fe-only hydrogenase, [(micro-SCH2)2NR]Fe2(CO)6 (, R=C6F4CF3-p; , R=C6H4CF3-p) and [(micro-SCH2)2NR]Fe2(CO)5(PPh3) (, R=C6F4CF3-p; , R=C6H4CF3-p), have been synthesized and well characterized. Spectroscopic and electrochemical studies demonstrate that the aryl-substituted complexes are stable toward a strong acid HBF4/Et2O, and electrocatalytic process for the hydrogen production is mostly dependent on the strength of the available proton source. When CH3COOH is used as the proton source, the electrocatalytic process begins with successively two one-electron reduction processes to produce H2 at Fe(0)Fe(0) (E2pc); whereas in the presence of strong acid, HBF4/Et2O, the process is initiated by protonation of a N-bridged atom followed by reduction of the protonated N-bridged atom around -1.29 V, and then release of H2 at Fe(0)Fe(I) (E1pc). Varying the strength of acid leads to the initial electron-transfer step from the reduction of a protonated N-bridged atom to the active site of [Fe(I)Fe(I)].

Catalytic Cycles for the Reduction of [UO2]2+ by Cytochrome c7 Proteins Proposed from DFT Calculations

The mechanism of the reduction of the hydrated uranyl cation, [UO2](2+), by the cytochromes G. sulfurreducens and D. acetoxidans has been studied using density functional theory calculations. We propose that the initial electron transfer step from the heme is to a cation-cation complex in the case of D. acetoxidans, but for G. sulfurreducens, it is to a single uranyl cation, which then forms a U(V)-U(VI) complex with a second uranyl cation. For both enzymes, the subsequent catalytic pathways are very similar. A U(V)-U(V) complex is formed, which then undergoes disproportionation via two successive protonation steps of one uranyl group, to give a U(VI)-U(IV) complex which dissociates to individual U(VI) and U(IV) species, the former being bound at the enzyme active site. Intermediate structures along the catalytic pathway are consistent with EXAFS data.