Showing papers by "Emiliano Fonda published in 2022"

Iron Insertion at the Assembly Site of the ISCU Scaffold Protein Is a Conserved Process Initiating Fe-S Cluster Biosynthesis.

Abstract: Iron-sulfur (Fe-S) clusters are prosthetic groups of proteins biosynthesized on scaffold proteins by highly conserved multi-protein machineries. Biosynthesis of Fe-S clusters into the ISCU scaffold protein is initiated by ferrous iron insertion, followed by sulfur acquisition, via a still elusive mechanism. Notably, whether iron initially binds to the ISCU cysteine-rich assembly site or to a cysteine-less auxiliary site via N/O ligands remains unclear. We show here by SEC, circular dichroism (CD), and Mössbauer spectroscopies that iron binds to the assembly site of the monomeric form of prokaryotic and eukaryotic ISCU proteins via either one or two cysteines, referred to the 1-Cys and 2-Cys forms, respectively. The latter predominated at pH 8.0 and correlated with the Fe-S cluster assembly activity, whereas the former increased at a more acidic pH, together with free iron, suggesting that it constitutes an intermediate of the iron insertion process. Iron not binding to the assembly site was non-specifically bound to the aggregated ISCU, ruling out the existence of a structurally defined auxiliary site in ISCU. Characterization of the 2-Cys form by site-directed mutagenesis, CD, NMR, X-ray absorption, Mössbauer, and electron paramagnetic resonance spectroscopies showed that the iron center is coordinated by four strictly conserved amino acids of the assembly site, Cys35, Asp37, Cys61, and His103, in a tetrahedral geometry. The sulfur receptor Cys104 was at a very close distance and apparently bound to the iron center when His103 was missing, which may enable iron-dependent sulfur acquisition. Altogether, these data provide the structural basis to elucidate the Fe-S cluster assembly process and establish that the initiation of Fe-S cluster biosynthesis by insertion of a ferrous iron in the assembly site of ISCU is a conserved mechanism.

Comparative Investigation of Red and Orange Roman Tesserae: Role of Cu and Pb in Colour Formation

Abstract: This study aims at the characterisation of red and orange glass tesserae from the 4th-century Roman villa of Noheda (Spain). Due to the limited number of analyses available for such ancient materials, many questions remain unanswered about the production processes in the Roman period. Six samples were chosen for their hue variations, including two samples showing banded patterns of red and orange. Differences in copper speciation were investigated by X-ray absorption spectroscopy and compared with colour and compositional variations obtained by EPMA. The shapes and sizes of colouring crystals could be investigated using scanning and transmission electron microscope imaging. The brown-red colour is due to metallic copper nano-particles and corresponds to a low-copper and low-lead group usually described in the literature. The orange samples and bands are coloured by copper oxide Cu2O nanoparticles with remaining Cu+ in the glass and have greater contents of Cu. Compositional analyses reveal that the same base glass is used in the red and orange bands with additions of Cu, Sn, Pb and probable Fe. Furthermore, based on our results and on the literature review, a high-copper low-lead group of glasses highlights the variability of compositions observed in cuprite colours.

Local environment of iodine dissolved as iodate in high-pressure aluminoborosilicate glasses: A I K-edge x-ray absorption spectroscopic study.

Abstract: The use of high-pressure synthesis conditions to produce I-bearing aluminoborosilicate represents a promising issue for the immobilization of 129I radioisotope. Furthermore, iodine appears to be more solubilized in glasses under its iodate (I5+) form rather than its iodide (I-) form. Currently, the local atomic environment for iodine is poorly constrained for I- and virtually unknown for I5+ or I7+. We used I K-edge x-ray absorption spectroscopy conducted at 20 K for determining the local atomic environment of iodine dissolved as I-, I5+, and I7+ in a series of aluminoborosilicate glasses. We determined that I- is surrounded by either Na+ or Ca2+ in agreement with previous studies. The signal collected from EXAFS reveals that I5+ is surrounded invariably by three oxygen atoms forming an IO3 - cluster charge compensated by Na+ and/or Ca2+. The I-O distance in iodate dissolved in glass is comparable to the I-O distance in crystalline compounds at ∼1.8 Å. The distance to the second nearest neighbor (Na+ or Ca2+) is also constant at ∼3.2 Å. This derived distance is identical to the distance between I- and Na+ or Ca2+ in the case of iodide local environment. For one sample containing iodate and periodate, the distinction between the local environment of I5+ and I7+ could not be made, suggesting that both environments have comparable EXAFS signals.

In Situ X-ray Absorption Spectroscopy Cells for High Pressure Homogeneous Catalysis

Abstract: We have designed, built, and tested two cells for in situ and, potentially, operando X-ray absorption spectroscopy experiments in transmission and fluorescence modes. The cells were developed for high-pressure and high-temperature conditions to study the catalytic processes under relevant industrial conditions. Operation of the cells was tested for Ru and Rh-based homogeneous and heterogeneous catalytic systems. Using synchrotron-based in situ X-ray absorption spectroscopy we tracked the evolution of active metal species during catalytic reactions. Our setup proved that it was capable to investigate liquid-state homogeneous and heterogenous systems under elevated temperatures, high pressures of reactive gasses, and in the presence of corrosive reagents.

Detection limit of next-generation of multi-element germanium detectors in the context of Environmental science

Abstract: One of the main challenges in Environmental sciences is the identification and chemical evolution of polluting traces (e.g, cadmium or antimony) in soil, which requires long acquistion times for accurate measurements at synchrotron facilities. In this context, the potential of a new generation multi-element germanium detectors to identify traces at 0.1-1 ppm in a reasonable time has been studied using Allpix Squared framework [1]. This code has been customized to include the three dimensional electric and weighting field maps generated by COMSOL Multiphysics software, and several features to model the sample environment at SOLEIL synchrotron and the signal response of a germanium detector equipped with a Digital Pulse Processor (DPP). The full simulation chain has been validated by experimental data from SAMBA beamline of SOLEIL synchrotron. This work presents a first estimation of the detection limit to cadmium traces in a soil sample for a future multi-element germanium detector, using this simulation chain.