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The progression of neurodegenerative disorders can lead to impaired neurotransmission; however, the role of pathogenic factors associated with these diseases and their impact on the structures and functions of neurotransmitters have not been clearly established. Here we report the discovery that conformational and functional changes of a native neuropeptide, somatostatin (SST), occur in the presence of copper ions, metal-free amyloid-β (Aβ) and metal-bound Aβ (metal-Aβ) found as pathological factors in the brains of patients with Alzheimer's disease. These pathological elements induce the self-assembly of SST and, consequently, prevent it from binding to the receptor. In the reverse direction, SST notably modifies the aggregation profiles of Aβ species in the presence of metal ions, attenuating their cytotoxicity and interactions with cell membranes. Our work demonstrates a loss of normal function of SST as a neurotransmitter and a gain of its modulative function against metal-Aβ under pathological conditions.

Methanotrophic bacteria utilize the nonheme diiron enzyme soluble methane monooxygenase (sMMO) to convert methane to methanol in the first step of their metabolic cycle under copper-limiting conditions. The structure of the sMMO Fe(IV)2 intermediate Q responsible for activating the inert C-H bond of methane (BDE = 104 kcal/mol) remains controversial, with recent studies suggesting both "open" and "closed" core geometries for its active site. In this study, we employ nuclear resonance vibrational spectroscopy (NRVS) to probe the geometric and electronic structure of intermediate Q at cryogenic temperatures. These data demonstrate that Q decays rapidly during the NRVS experiment. Combining data from several years of measurements, we derive the NRVS vibrational features of intermediate Q as well as its cryoreduced decay product. A library of 90 open and closed core models of intermediate Q is generated using density functional theory to analyze the NRVS data of Q and its cryoreduced product as well as prior spectroscopic data on Q. Our analysis reveals that a subset of closed core models reproduce these newly acquired NRVS data as well as prior data. The reaction coordinate with methane is also evaluated using both closed and open core models of Q. These studies show that the potent reactivity of Q toward methane resides in the "spectator oxo" of its Fe(IV)2O2 core, in contrast to nonheme mononuclear Fe(IV)═O enzyme intermediates that H atoms abstract from weaker C-H bonds.

A simpler and less expensive fabrication process is one of the essential demands for the commercialization of perovskite solar cells (PeSCs). Especially, inverted PeSCs (I-PeSCs) require a cathode buffer layer (CBL) for fabricating highly efficient and stable PeSCs. However, this increases the number of fabrication step. Here, we demonstrate highly stable and efficient cathode-buffer-layer-free I-PeSCs via additive engineering on an ETL, which is based on phenyl-C61-butyric acid methyl ester (PC61BM) with a small amount of poly(methyl methacrylate) (PMMA). This modified ETL shows not only a simplified fabrication process but also effective extraction of charge from the perovskite to a high work function copper electrode (Cu) by formation of an interfacial dipole at the interfaces between the ETL and the Cu. Additionally, it exhibits good passivation of the trap density existing along the grain boundaries and surface of the perovskite layer, reducing the non-radiative recombination and consistent with the increases in open-circuit voltage (Voc). As a result, I-PeSCs with a blend PC61BM : PMMA ETL demonstrate an enhancement in the power conversion efficiency (PCE) from 13.55% (without PMMA) to 18.38%. Furthermore, they exhibit both burn-in-free behaviour in photostability measurements by maximum power-point tracking (MPPT) method and long-term air-stability (30 days for T90) in ambient air. Lastly, we obtained PCE of 15.03% and 16.83% for large-area (1 cm2) I-PeSCs with PC61BM and PC61BM : PMMA, respectively. This method provides an alternative route to reduce the fabrication time and budget for commercialization of I-PeSCs without sacrificing device performance.

Photoexcitation is one of the acknowledged methods to activate Ni-based cross-coupling reactions, but factors that govern the photoactivity of organonickel complexes have not yet been established. Here we report the excited-state cross-coupling activities of Ni(ii) metallacycle compounds, which display ∼104 times enhancement for the C–S bond-forming reductive elimination reaction upon Ni-centered ligand-field transitions. The effects of excitation energy and ancillary ligands on photoactivity have been investigated with 17 different nickelacycle species in combination with four corresponding acyclic complexes. Spectroscopic and computational electronic structural characterizations reveal that, regardless of coordinated species, d–d transitions can induce Ni–C bond homolysis, and that the reactivity of the resulting Ni(i) species determines the products of the overall reaction. The photoactivity mechanism established in this study provides general insights into the excited-state chemistry of organonickel(ii) complexes., d–d excitations can accelerate C–S reductive eliminations of nickelacycles via intersystem crossing to a repulsive 3(C-to-Ni charge transfer) state inducing Ni–C bond homolysis. This homolytic photoreactivity is common for organonickel(ii) complexes.

The excited-state reductive elimination (RE) activities of a metal–metal bonded PdIII2 complex, [(2-phenylpyridyl)Pd(μ-acetate)Cl]2, are described. The C–Cl bond-forming RE reaction is accelerated by up to five orders of magnitude upon visible photoexcitation, which induces the Pd–Pd bond dissociation. This ligand field variation reduces the energy cost for intramolecular charge transfer (ICT) from coupling substrate to the metal center involved in the RE reaction. The correlation found between the ligand field and the RE activity indicates the ICT energy as a fundamental descriptor for RE reactions.

Regeneration of nicotinamide adenine dinucleotide (NADH) has been the primary interest in the field of enzymatic transformation, especially associating oxidoreductases given the stoichiometric consumption. The synthesized carbene-ligated rhodium complex [(η

The α-ketoglutarate (αKG)-dependent oxygenases catalyze a diverse range of chemical reactions using a common high-spin FeIV═O intermediate that, in most reactions, abstract a hydrogen atom from the substrate. Previously, the FeIV═O intermediate in the αKG-dependent halogenase SyrB2 was characterized by nuclear resonance vibrational spectroscopy (NRVS) and density functional theory (DFT) calculations, which demonstrated that it has a trigonal-pyramidal geometry with the scissile C-H bond of the substrate calculated to be perpendicular to the Fe-O bond. Here, we have used NRVS and DFT calculations to show that the FeIV═O complex in taurine dioxygenase (TauD), the αKG-dependent hydroxylase in which this intermediate was first characterized, also has a trigonal bipyramidal geometry but with an aspartate residue replacing the equatorial halide of the SyrB2 intermediate. Computational analysis of hydrogen atom abstraction by square pyramidal, trigonal bipyramidal, and six-coordinate FeIV═O complexes in two different substrate orientations (one more along [σ channel] and another more perpendicular [π channel] to the Fe-O bond) reveals similar activation barriers. Thus, both substrate approaches to all three geometries are competent in hydrogen atom abstraction. The equivalence in reactivity between the two substrate orientations arises from compensation of the promotion energy (electronic excitation within the d manifold) required to access the π channel by the significantly larger oxyl character present in the pπ orbital oriented toward the substrate, which leads to an earlier transition state along the C-H coordinate.

This paper presents an automatic proficiency assessment method for a non‐native Korean read utterance using bidirectional long short–term memory (BLSTM)–based acoustic models (AMs) and speech data augmentation techniques. Specifically, the proposed method considers two scenarios, with and without prompted text. The proposed method with the prompted text performs (a) a speech feature extraction step, (b) a forced‐alignment step using a native AM and non‐native AM, and (c) a linear regression–based proficiency scoring step for the five proficiency scores. Meanwhile, the proposed method without the prompted text additionally performs Korean speech recognition and a subword un‐segmentation for the missing text. The experimental results indicate that the proposed method with prompted text improves the performance for all scores when compared to a method employing conventional AMs. In addition, the proposed method without the prompted text has a fluency score performance comparable to that of the method with prompted text.

High-valent Ni complexes have proven to be good platforms for diverse cross-coupling reactions that are otherwise difficult to be achieved with conventional low-valent catalysts. However, their reductive elimination (RE) activities are still significantly variable by up to 5 orders of magnitude, depending on the supporting ligand and oxidation state of the Ni center. To elucidate frontier molecular orbitals (FMOs) that determine the RE activity of the Ni center, the electronic structures of cycloneophyl (CH

Along with the mitigation of CO2 emission, recently, the CO2-derived formic acid process has drawn attention as a promising platform for the renewable-energy-derived hydrogen storage cycle by using formic acid as a liquid organic hydrogen carrier (LOHC). Here, a heterogenized Ru molecular catalyst on a bpyTN-30-CTF support is prepared and successfully implemented in an integrated trickle-bed reactor system for continuous CO2 hydrogenation to produce formic acid. The bpyTN-30-CTF support with an alternative structure of the bpy and TN motif increases the porosity and metal anchoring sites. The Ru/bpyTN-30-CTF catalyst prepared using the bpyTN-30-CTF support displays sufficient catalytic activity for commercialization. Under the continuous process, the catalyst exhibits substantial catalytic performance with the highest productivity of 669.0 gform. gcat−1 d−1 with CO2 conversion of 44.8% for a superficial gas velocity of 72 cm s−1. Furthermore, the catalyst shows excellent stability in the continuous hydrogenation process with a trickle-bed reactor over 30 days of operation, reaching a total turnover number of 524 000 without any significant deactivation. Based on kinetic data, a new process to produce formic acid by CO2 hydrogenation has thus been proposed here.

Methanotrophic bacteria utilize the non-heme diiron enzyme soluble methane monooxygenase (sMMO) to convert methane to methanol in the first step of their metabolic cycle under copper-limiting conditions. The structure of the sMMO Fe(IV)(2) intermediate Q responsible for activating the inert C-H bond of methane (BDE = 104 kcal/mol) remains controversial, with recent studies suggesting both “open” and “closed” core geometries for its active site. In this study, we employ nuclear resonance vibrational spectroscopy (NRVS) to probe the geometric and electronic structure of intermediate Q at cryogenic temperatures. These data demonstrate that Q decays rapidly during the NRVS experiment. Combining data from several years of measurements, we derive the NRVS vibrational features of intermediate Q as well as its cryoreduced decay product. A library of 90 open and closed core models of intermediate Q is generated using density functional theory (DFT) to analyze the NRVS data of Q and its cryoreduced product, as well as prior spectroscopic data on Q. Our analysis reveals that a subset of closed core models reproduce these newly acquired NRVS data as well as prior data. The reaction coordinate with methane is also evaluated using both closed and open core models of Q. These studies show that the potent reactivity of Q towards methane resides in the “spectator oxo” of its Fe(IV)(2)O(2) core, in contrast to non-heme mononuclear Fe(IV)=O enzyme intermediates that H-atom abstract from weaker C-H bonds.

Molybdenum sulfides have attracted widespread attention as promising nonprecious-metal catalysts for the hydrogen evolution reaction (HER). Since the MoS2 edge was proposed as a major active site, ...

The α-ketoglutarate (αKG)-dependent oxygenases catalyze a diverse range of chemical reactions using a common high-spin Fe(IV)=O intermediate and, in most reactions, abstracts a hydrogen atom from the substrate. Previously, the Fe(IV)=O intermediate in the αKG-dependent halogenase SyrB2 was characterized by nuclear resonance vibrational spectroscopy (NRVS) and density functional theory (DFT) calculations, which demonstrated that it has a trigonal-pyramidal geometry with the scissile C-H bond of the substrate calculated to be perpendicular to the Fe-O bond. Here, we have used NRVS and DFT calculations to show that the Fe(IV)=O complex in taurine dioxygenase (TauD), the αKG-dependent hydroxylase in which this intermediate was first characterized, also has a trigonal bipyramidal geometry, but with an aspartate residue replacing the equatorial halide of the SyrB2 intermediate. Computational analysis of hydrogen atom abstraction by square pyramidal, trigonal bipyramidal, and six-coordinate Fe(IV)=O complexes in two different substrate orientations (one more along [σ channel] and another more perpendicular [π channel] to the Fe-O bond) reveals similar activation barriers. Thus, both substrate approaches to all three geometries are competent in hydrogen atom abstraction. The equivalence in reactivity between the two substrate orientations arises from compensation of the promotion energy (electronic excitation within the d manifold) required to access the π channel by the significantly larger oxyl character present in the pπ orbital oriented towards the substrate, which leads to an earlier transition state along the C-H coordinate.

This paper aims to design an online, low-latency, and high-performance speech recognition system using a bidirectional long short-term memory (BLSTM) acoustic model. To achieve this, we adopt a server-client model and a context-sensitive-chunk-based approach. The speech recognition server manages a main thread and a decoder thread for each client and one worker thread. The main thread communicates with the connected client, extracts speech features, and buffers the features. The decoder thread performs speech recognition, including the proposed multichannel parallel acoustic score computation of a BLSTM acoustic model, the proposed deep neural network-based voice activity detector, and Viterbi decoding. The proposed acoustic score computation method estimates the acoustic scores of a context-sensitive-chunk BLSTM acoustic model for the batched speech features from concurrent clients, using the worker thread. The proposed deep neural network-based voice activity detector detects short pauses in the utterance to reduce response latency, while the user utters long sentences. From the experiments of Korean speech recognition, the number of concurrent clients is increased from 22 to 44 using the proposed acoustic score computation. When combined with the frame skipping method, the number is further increased up to 59 clients with a small accuracy degradation. Moreover, the average user-perceived latency is reduced from 11.71 s to 3.09&ndash, 5.41 s by using the proposed deep neural network-based voice activity detector.

A new series of ITIC-based non-fullerene acceptors (NFAs), namely m-ITTC-Oeh-SM (IT-SM) and m-ITTC-Oeh-DSM (IT-DSM), which contain thiophene-substituted end groups (EGs) with single and double thiomethyl side chains, respectively, were designed, synthesized, and characterized in this study. The IT-SM NFA exhibited a narrower optical band gap, redshifted absorption, stronger crystallinity, and smoother surface morphology than those of the IT-DSM NFA. Organic solar cells (OSCs) based on the polymer donor PBDB-T and IT-SM reported a power conversion efficiency (PCE) of 8.98% with the addition of 0.5 v/v% 1,8-diiodooctane (DIO), whereas the PCE of the PBDB-T:IT-DSM device was equal to 3.02%. The difference between the PCE values of these two devices was attributed to the gap of the short-circuit current and fill factor. Detailed analyses of the charge transport, collection, and morphology of the blend films were performed to investigate the difference between the device performances. The results of this study can help formulate detailed guidelines for the commercialized production of NFAs.

Translation initiation factor eIF4E forms eIF4E-eIF4G complex at the 5' cap of mRNA. This interaction can be inhibited by the family of 4E-binding proteins (4E-BP). In yeast Saccharomyces cerevisiae, two 4E-BPs, Caf20 and Eap1, compete with eIF4G for binding to eIF4E via the shared conserved interaction motif. In order to investigate the roles of Caf20 in gene-specific translational regulation and the formation of mRNA granules (P-bodies), we introduced substitution mutations, caf20-Y4A or caf20-L9A, in the eIF4E-binding motif for CAF20. Overexpression of the wild-type CAF20 showed an increased protein level of Ste12 transcription factor as well as highly developed P-body formation. However, 4E-binding site mutations of CAF20 led to a reduced number of P-body foci and decreased levels of Ste12 protein. The phenotypes of the caf20 deletion mutation were also analyzed, and we suggest that Caf20 plays a critical role in Ste12 protein expression and in the control of P-body formation.

One bottleneck in the realization of CO2 conversion into value-added compounds is the lack of catalysts with both excellent activity and recyclability. Herein, a catalyst is designed for the hydrogenation of CO2 to formate to boost up these features by considering the leaching pathway of previously reported heterogenized catalyst; the design strategy incorporates oxyanionic ligand(s) in the coordination sphere to provide a pathway for both preventing the deleterious interactions and assisting the heterolysis of H2. The tailored heterogenized catalyst, [bpy-CTF-Ru(acac)2]Cl, demonstrated excellent recyclability over consecutive runs with a highest turnover frequency of 22 700 h–1, and produced a highest formate concentration of 1.8 M in 3 h. This work is significant in elucidating new principles for the development of industrially viable hydrogenation catalysts.

Binuclear non-heme iron enzymes activate O2 for diverse chemistries that include oxygenation of organic substrates and hydrogen atom abstraction. This process often involves the formation of peroxo-bridged biferric intermediates, only some of which can perform electrophilic reactions. To elucidate the geometric and electronic structural requirements to activate peroxo reactivity, the active peroxo intermediate in 4-aminobenzoate N-oxygenase (AurF) has been characterized spectroscopically and computationally. A magnetic circular dichroism study of reduced AurF shows that its electronic and geometric structures are poised to react rapidly with O2. Nuclear resonance vibrational spectroscopic definition of the peroxo intermediate formed in this reaction shows that the active intermediate has a protonated peroxo bridge. Density functional theory computations on the structure established here show that the protonation activates peroxide for electrophilic/single-electron-transfer reactivity. This activation of peroxide by protonation is likely also relevant to the reactive peroxo intermediates in other binuclear non-heme iron enzymes.

Metal(TMC) complexes (TMC = tetramethylcyclam) exclusively form trans-I diastereoisomers; thus, little is known about the trans-III isomers. Herein, we report a new method to prepare the trans-III-[Cu(TMC)]2+ complex via an anion effect. Our crystallographic, spectroscopic, and computational analyses validated that [Cu(TMC)](NO3)2 produced the trans-III complex, different from the trans-I structure of [Cu(TMC)](ClO4)2.

Strongly emissive cuprous complexes containing a stable σ-SiH-Cu motif were prepared. These complexes serve as a proof of principle that σ-complexation can be utilized as a design principle for engineering responsive light emitting materials. Their excited state lifetimes were found to be long (∼20 μs) with high quantum yields (φ = 0.40-0.59).

We investigated three conjugated polymers, PSBDB-T, P1SBDB-TS, and PSBDB-TS, having different numbers of alkylthio groups. The polymers showed a wide bandgap of 1.76–1.81 eV with broad absorption range from 300–700 nm and were used as donor polymers in organic photovoltaic (OPV) devices with the non-fullerene acceptor, IT-M. By increasing the number of alkythio groups in their chemical structures, the HOMO levels of the polymers down-shifted as −5.46 eV, −5.51 eV and −5.56 eV, and consequently, the VOC of the OPV cells gradually increased as 0.91 V, 0.92 V and 0.94 V for PSBDB-T, P1SBDB-TS, and PSBDB-TS, respectively. P1SBDB-TS having three alkylthio groups (two for benzo[1,2-b:4,5-b']dithiophene and one for benzo[1,2-c:4,5-c’]dithiophene-4,8-dione unit) showed the best power conversion efficiency of 6.88% among three polymers due to its high crystallinity and low bimolecular recombination in bulk heterojunction system.

The Rieske dioxygenases are a major subclass of mononuclear nonheme iron enzymes that play an important role in bioremediation. Recently, a high-spin FeIII–(hydro)-peroxy intermediate (BZDOp) has been trapped in the peroxide shunt reaction of benzoate 1,2-dioxygenase. Defining the structure of this intermediate is essential to understanding the reactivity of these enzymes. Nuclear resonance vibrational spectroscopy (NRVS) is a recently developed synchrotron technique that is ideal for obtaining vibrational, and thus structural, information on Fe sites, as it gives complete information on all vibrational normal modes containing Fe displacement. In this study, we present NRVS data on BZDOp and assign its structure using these data coupled to experimentally calibrated density functional theory calculations. From this NRVS structure, we define the mechanism for the peroxide shunt reaction. The relevance of the peroxide shunt to the native FeII/O2 reaction is evaluated. For the native FeII/O2 reaction, an FeIII–superoxo intermediate is found to react directly with substrate. This process, while uphill thermodynamically, is found to be driven by the highly favorable thermodynamics of proton-coupled electron transfer with an electron provided by the Rieske [2Fe-2S] center at a later step in the reaction. These results offer important insight into the relative reactivities of FeIII–superoxo and FeIII–hydroperoxo species in nonheme Fe biochemistry.

Three distinct families of ATP:corrinoid adenosyltransferases (ACATs) exist that are capable of converting vitamin B12 derivatives to coenzyme B12 by catalyzing the thermodynamically challenging reduction of Co(II)rrinoids to form “super-nucleophilic” Co(I) intermediates. While the structures and mechanisms for the other two ACAT families have been studied extensively, little is known about the EutT enzymes beyond the fact that they exhibit a unique requirement for a divalent metal cofactor for enzymatic activity. In this study we have obtained compelling evidence that EutT converts cob(II)alamin to an effectively four-coordinate Co(II) species so as to facilitate Co(II) → Co(I) reduction. Intriguingly, EutT fails to promote axial ligand dissociation for the substrate analogue cob(II)inamide, a natural precursor of cob(II)alamin. This unique substrate specificity of EutT has important physiological implications.

Nuclear resonance vibrational spectroscopy (NRVS) is a powerful technique that can provide geometric structural information on key reaction intermediates of Fe-containing systems when utilized in combination with density functional theory (DFT). However, in the case of binuclear nonheme iron enzymes, DFT-predicted NRVS spectra have been found to be sensitive to the truncation method used to model the active sites of the enzymes. Therefore, in this study various-level truncation schemes have been tested to predict the NRVS spectrum of a binuclear nonheme iron enzyme, and a reasonably sized DFT model that is suitable for employing the NRVS/DFT combined methodology to characterize binuclear nonheme iron enzymes has been developed.

An amyloidogenic peptide, amyloid-β (Aβ), has been implicated as a contributor to the neurotoxicity of Alzheimer’s disease (AD) that continues to present a major socioeconomic burden for our society. Recently, the use of metal complexes capable of cleaving peptides has arisen as an efficient tactic for amyloid management; unfortunately, little has been reported to pursue this strategy. Herein, we report a novel approach to validate the hydrolytic cleavage of divalent metal complexes toward two major isoforms of Aβ (Aβ40 and Aβ42) and tune their proteolytic activity based on the choice of metal centers (M = Co, Ni, Cu, and Zn) which could be correlated to their anti-amyloidogenic properties. Such metal-dependent tunability was facilitated employing a tetra-N-methylated cyclam (TMC) ligand that imparts unique geometric and stereochemical control, which has not been available in previous systems. Co(II)(TMC) was identified to noticeably cleave Aβ peptides and control their aggregation, reporting the first Co...

Mononuclear non-haem iron (NHFe) enzymes catalyse a broad range of oxidative reactions, including halogenation, hydroxylation, ring closure, desaturation and aromatic ring cleavage reactions. They are involved in a number of biological processes, including phenylalanine metabolism, the production of neurotransmitters, the hypoxic response and the biosynthesis of secondary metabolites. The reactive intermediate in the catalytic cycles of these enzymes is a high-spin S = 2 Fe(IV)=O species, which has been trapped for a number of NHFe enzymes, including the halogenase SyrB2 (syringomycin biosynthesis enzyme 2). Computational studies aimed at understanding the reactivity of this Fe(IV)=O intermediate are limited in applicability owing to the paucity of experimental knowledge about its geometric and electronic structure. Synchrotron-based nuclear resonance vibrational spectroscopy (NRVS) is a sensitive and effective method that defines the dependence of the vibrational modes involving Fe on the nature of the Fe(IV)=O active site. Here we present NRVS structural characterization of the reactive Fe(IV)=O intermediate of a NHFe enzyme, namely the halogenase SyrB2 from the bacterium Pseudomonas syringae pv. syringae. This intermediate reacts via an initial hydrogen-atom abstraction step, performing subsequent halogenation of the native substrate or hydroxylation of non-native substrates. A correlation of the experimental NRVS data to electronic structure calculations indicates that the substrate directs the orientation of the Fe(IV)=O intermediate, presenting specific frontier molecular orbitals that can activate either selective halogenation or hydroxylation.

While the geometric and electronic structures of vitamin B12 (cyanocobalamin, CNCbl) and its reduced derivatives Co(2+)cobalamin (Co(2+)Cbl) and Co(1+)cobalamin (Co(1+)Cbl(-)) are now reasonably well established, their vibrational properties, in particular their resonance Raman (rR) spectra, have remained quite poorly understood. The goal of this study was to establish definitive assignments of the corrin-based vibrational modes that dominate the rR spectra of vitamin B12 in its Co(3+), Co(2+), and Co(1+) oxidation states. rR spectra were collected for all three species with laser excitation in resonance with the most intense corrin-based π → π* transitions. These experimental data were used to validate the computed vibrational frequencies, eigenvector compositions, and relative rR intensities of the normal modes of interest as obtained by density functional theory (DFT) calculations. Importantly, the computational methodology employed in this study successfully reproduces the experimental observation that the frequencies and rR excitation profiles of the corrin-based vibrational modes vary significantly as a function of the cobalt oxidation state. Our DFT results suggest that this variation reflects large differences in the degree of mixing between the occupied Co 3d orbitals and empty corrin π* orbitals in CNCbl, Co(2+)Cbl, and Co(1+)Cbl(-). As a result, vibrations mainly involving stretching of conjugated C-C and C-N bonds oriented along one axis of the corrin ring may, in fact, couple to a perpendicularly polarized electronic transition. This unusual coupling between electronic transitions and vibrational motions of corrinoids greatly complicates an assignment of the corrin-based normal modes of vibrations on the basis of their rR excitation profiles.

The human-type ATP:corrinoid adenosyltransferase PduO from Lactobacillus reuteri (LrPduO) catalyzes the adenosylation of Co(II)rrinoids to generate adenosylcobalamin (AdoCbl) or adenosylcobinamide (AdoCbi(+)). This process requires the formation of "supernucleophilic" Co(I)rrinoid intermediates in the enzyme active site which are properly positioned to abstract the adeonsyl moiety from co-substrate ATP. Previous magnetic circular dichroism (MCD) spectroscopic and X-ray crystallographic analyses revealed that LrPduO achieves the thermodynamically challenging reduction of Co(II)rrinoids by displacing the axial ligand with a non-coordinating phenylalanine residue to produce a four-coordinate species. However, relatively little is currently known about the interaction between the tetradentate equatorial ligand of Co(II)rrinoids (the corrin ring) and the enzyme active site. To address this issue, we have collected resonance Raman (rR) data of Co(II)rrinoids free in solution and bound to the LrPduO active site. The relevant resonance-enhanced vibrational features of the free Co(II)rrinoids are assigned on the basis of rR intensity calculations using density functional theory to establish a suitable framework for interpreting rR spectral changes that occur upon Co(II)rrinoid binding to the LrPduO/ATP complex in terms of structural perturbations of the corrin ring. To complement our rR data, we have also obtained MCD spectra of Co(II)rrinoids bound to LrPduO complexed with the ATP analogue UTP. Collectively, our results provide compelling evidence that in the LrPduO active site, the corrin ring of Co(II)rrinoids is firmly locked in place by several amino acid side chains so as to facilitate the dissociation of the axial ligand.