The tetranuclear copper active site of nitrous oxide reductase: the CuZ center

J Biol Inorg Chem (2011) 16:183–194 DOI 10.1007/s00775-011-0753-3This review focuses on the novel CuZ center of nitrous oxide reductase, an important enzyme owing to the environmental significance of the reaction it catalyzes, reduction of nitrous oxide, and the unusual nature of its catalytic center, named CuZ. The structure of the CuZ center, the unique tetranuclear copper center found in this enzyme, opened a novel area of research in metallobiochemistry. In the last decade, there has been progress in defining the structure of the CuZ center, characterizing the mechanism of nitrous oxide reduction, and identifying intermediates of this reaction. In addition, the determination of the structure of the CuZ center allowed a structural interpretation of the spectroscopic data, which was supported by theoretical calculations. The current knowledge of the structure, function, and spectroscopic characterization of the CuZ center is described here. We would like to stress that although many questions have been answered, the CuZ center remains a scientific challenge, with many hypotheses still being formed

The electron transfer complex between nitrous oxide reductase and its electron donors

J Biol Inorg Chem (2011) 16:1241–1254 DOI 10.1007/s00775-011-0812-9Identifying redox partners and the interaction surfaces is crucial for fully understanding electron flow in a respiratory chain. In this study, we focused on the interaction of nitrous oxide reductase (N2OR), which catalyzes the final step in bacterial denitrification, with its physiological electron donor, either a c-type cytochrome or a type 1 copper protein. The comparison between the interaction of N2OR from three different microorganisms, Pseudomonas nautica, Paracoccus denitrificans, and Achromobacter cycloclastes, with their physiological electron donors was performed through the analysis of the primary sequence alignment, electrostatic surface, and molecular docking simulations, using the bimolecular complex generation with global evaluation and ranking algorithm. The docking results were analyzed taking into account the experimental data, since the interaction is suggested to have either a hydrophobic nature, in the case of P. nautica N2OR, or an electrostatic nature, in the case of P. denitrificans N2OR and A. cycloclastes N2OR. A set of well-conserved residues on the N2OR surface were identified as being part of the electron transfer pathway from the redox partner to N2OR(Ala495, Asp519, Val524, His566 and Leu568 numbered according to the P. nautica N2OR sequence). Moreover, we built a model for Wolinella succinogenes N2OR, an enzyme that has an additional c-type-heme-containing domain. The structures of the N2OR domain and the c-type-heme-containing domain were modeled and the full-length structure was obtained by molecular docking simulation of these two domains. The orientation of the c-type-heme-containing domain relative to the N2OR domain is similar to that found in the other electron transfer complexes

Motion of wave fronts in semiconductor superlattices

An analysis of wave front motion in weakly coupled doped semiconductor superlattices is presented. If a dimensionless doping is sufficiently large, the superlattice behaves as a discrete system presenting front propagation failure and the wave fronts can be described near the threshold currents J(i) (i= 1,2) at which they depin and move. The wave front velocity scales with current as \J-J(i)\(1/2). If the dimensionless doping is low enough, the superlattice behaves as a continuum system and wave fronts are essentially shock waves whose velocity obeys an equal area rule

Electron transfer complex between nitrous oxide reductase and cytochrome c552 from Pseudomonas nautica: kinetic, nuclear magnetic resonance, and docking studies

Biochemistry. 2008 Oct 14;47(41):10852-62. doi: 10.1021/bi801375qThe multicopper enzyme nitrous oxide reductase (N 2OR) catalyzes the final step of denitrification, the two-electron reduction of N 2O to N 2. This enzyme is a functional homodimer containing two different multicopper sites: CuA and CuZ. CuA is a binuclear copper site that transfers electrons to the tetranuclear copper sulfide CuZ, the catalytic site. In this study, Pseudomonas nautica cytochrome c 552 was identified as the physiological electron donor. The kinetic data show differences when physiological and artificial electron donors are compared [cytochrome vs methylviologen (MV)]. In the presence of cytochrome c 552, the reaction rate is dependent on the ET reaction and independent of the N 2O concentration. With MV, electron donation is faster than substrate reduction. From the study of cytochrome c 552 concentration dependence, we estimate the following kinetic parameters: K m c 552 = 50.2 +/- 9.0 muM and V max c 552 = 1.8 +/- 0.6 units/mg. The N 2O concentration dependence indicates a K mN 2 O of 14.0 +/- 2.9 muM using MV as the electron donor. The pH effect on the kinetic parameters is different when MV or cytochrome c 552 is used as the electron donor (p K a = 6.6 or 8.3, respectively). The kinetic study also revealed the hydrophobic nature of the interaction, and direct electron transfer studies showed that CuA is the center that receives electrons from the physiological electron donor. The formation of the electron transfer complex was observed by (1)H NMR protein-protein titrations and was modeled with a molecular docking program (BiGGER). The proposed docked complexes corroborated the ET studies giving a large number of solutions in which cytochrome c 552 is placed near a hydrophobic patch located around the CuA center

HySenS data exploitation for urban land cover analysis

This paper addresses the use of HySenS airborne hyperspectral data for environmental urban monitoring. It is known that hyperspectral data can help to characterize some of the relations between soil composition, vegetation characteristics, and natural/artificial materials in urbanized areas. During the project we collected DAIS and ROSIS data over the urban test area of Pavia, Northern Italy, though due to a late delivery of ROSIS data only DAIS data was used in this work. Here we show results referring to an accurate characterization and classification of land cover/use, using different supervised approaches, exploiting spectral as well as spatial information. We demonstrate the possibility to extract from the hyperspectral data information which is very useful for environmental characterization of urban areas

A new CuZ active form in the catalytic reduction of N2O by nitrous oxide reductase from Pseudomonas nautica

J Biol Inorg Chem (2010) 15:967–976 DOI 10.1007/s00775-010-0658-6The final step of bacterial denitrification, the two-electron reduction of N2O to N2, is catalyzed by a multi-copper enzyme named nitrous oxide reductase. The catalytic centre of this enzyme is a tetranuclear copper site called CuZ, unique in biological systems. The in vitro reconstruction of the activity requires a slow activation in the presence of the artificial electron donor, reduced methyl viologen, necessary to reduce CuZ from the resting non-active state (1CuII/3CuI) to the fully reduced state (4CuI), in contrast to the turnover cycle, which is very fast. In the present work, the direct reaction of the activated form of Pseudomonas nautica nitrous oxide reductase with stoichiometric amounts of N2O allowed the identification of a new reactive intermediate of the catalytic centre, CuZ°, in the turnover cycle, characterized by an intense absorption band at 680 nm. Moreover, the first mediated electrochemical study of Ps. nautica nitrous oxide reductase with its physiological electron donor, cytochrome c-552, was performed. The intermolecular electron transfer was analysed by cyclic voltammetry, under catalytic conditions, and a second-order rate constant of (5.5 ± 0.9) × 105 M−1 s−1 was determined. Both the reaction of stoichiometric amounts of substrate and the electrochemical studies show that the active CuZ° species, generated in the absence of reductants, can rearrange to the resting non-active CuZ state. In this light, new aspects of the catalytic and activation/inactivation mechanism of the enzyme are discussed

HySenS data exploitation for urban land cover analysis

This paper addresses the use of HySenS airborne hyperspectral data for environmental urban monitoring. It is known that hyperspectral data can help to characterize some of the relations between soil composition, vegetation characteristics, and natural/artificial materials in urbanized areas. During the project we collected DAIS and ROSIS data over the urban test area of Pavia, Northern Italy, though due to a late delivery of ROSIS data only DAIS data was used in this work. Here we show results referring to an accurate characterization and classification of land cover/use, using different supervised approaches, exploiting spectral as well as spatial information. We demonstrate the possibility to extract from the hyperspectral data information which is very useful for environmental characterization of urban areas

Neural correlates of the rubber hand illusion in amputees: a report of two cases

One of the current challenges in the field of advanced prosthetics is the development of artificial limbs that provide the user with detailed sensory feedback. Sensory feedback from our limbs is not only important for proprioceptive awareness and motor control, but also essential for providing us with a feeling of ownership or simply put, the sensation that our limbs actually belong to ourselves. The strong link between sensory feedback and ownership has been repeatedly demonstrated with the so-called rubber hand illusion (RHI), during which individuals are induced with the illusory sensation that an artificial hand is their own. In healthy participants, this occurs via integration of visual and tactile signals, which is primarily supported by multisensory regions in premotor and intraparietal cortices. Here, we describe a functional magnetic resonance imaging (fMRI) study with two upper limb amputees, showing for the first time that the same brain regions underlie ownership sensations of an artificial hand in this population. Albeit preliminary, these findings are interesting from both a theoretical as well as a clinical point of view. From a theoretical perspective, they imply that even years after the amputation, a few seconds of synchronous visuotactile stimulation are sufficient to activate hand-centered multisensory integration mechanisms. From a clinical perspective, they show that a very basic sensation of touch from an artificial hand can be obtained by simple but precisely targeted stimulation of the stump, and suggest that a similar mechanism implemented in prosthetic hands would greatly facilitate ownership sensations and in turn, acceptance of the prosthesis

Telephone conversation impairs sustained visual attention via a central bottleneck

Recent research has shown that holding telephone conversations disrupts one's driving ability. We asked whether this effect could be attributed to a visual attention impairment. In Experiment 1, participants conversed on a telephone or listened to a narrative while engaged in multiple object tracking (MOT), a task requiring sustained visual attention. We found that MOT was disrupted in the telephone conversation condition, relative to single-task MOT performance, but that listening to a narrative had no effect. In Experiment 2, we asked which component of conversation might be interfering with MOT performance. We replicated the conversation and single-task conditions of Experiment 1 and added two conditions in which participants heard a sequence of words over a telephone. In the shadowing condition, participants simply repeated each word in the sequence. In the generation condition, participants were asked to generate a new word based on each word in the sequence. Word generation interfered with MOT performance, but shadowing did not. The data indicate that telephone conversation disrupts attention at a central stage, the act of generating verbal stimuli, rather than at a peripheral stage, such as listening or speaking

Diffusion tensor imaging of Parkinson's disease, multiple system atrophy and progressive supranuclear palsy: a tract-based spatial statistics study

Although often clinically indistinguishable in the early stages, Parkinson's disease (PD), Multiple System Atrophy (MSA) and Progressive Supranuclear Palsy (PSP) have distinct neuropathological changes. The aim of the current study was to identify white matter tract neurodegeneration characteristic of each of the three syndromes. Tract-based spatial statistics (TBSS) was used to perform a whole-brain automated analysis of diffusion tensor imaging (DTI) data to compare differences in fractional anisotropy (FA) and mean diffusivity (MD) between the three clinical groups and healthy control subjects. Further analyses were conducted to assess the relationship between these putative indices of white matter microstructure and clinical measures of disease severity and symptoms. In PSP, relative to controls, changes in DTI indices consistent with white matter tract degeneration were identified in the corpus callosum, corona radiata, corticospinal tract, superior longitudinal fasciculus, anterior thalamic radiation, superior cerebellar peduncle, medial lemniscus, retrolenticular and anterior limb of the internal capsule, cerebral peduncle and external capsule bilaterally, as well as the left posterior limb of the internal capsule and the right posterior thalamic radiation. MSA patients also displayed differences in the body of the corpus callosum corticospinal tract, cerebellar peduncle, medial lemniscus, anterior and superior corona radiata, posterior limb of the internal capsule external capsule and cerebral peduncle bilaterally, as well as the left anterior limb of the internal capsule and the left anterior thalamic radiation. No significant white matter abnormalities were observed in the PD group. Across groups, MD correlated positively with disease severity in all major white matter tracts. These results show widespread changes in white matter tracts in both PSP and MSA patients, even at a mid-point in the disease process, which are not found in patients with PD