Biomedical subjects

Barbara Fayard

Publications and source records attributed to Barbara Fayard.

5 recordsLinked to original sources

PubMed2005

Packing of the prion Ure2p in protein fibrils probed by fluorescence X-ray near-edge structure spectroscopy at sulfur K-edge.

The soluble protein Ure2p from the yeast Saccharomyces cerevisiae assembles in vitro into straight and insoluble protein fibrils, through subtle changes of conformation. Whereas the structure of soluble Ure2p has been revealed by X-ray crystallography, further characterization of the structure of insoluble Ure2p fibrils is needed. We performed X-ray absorption near-edge spectroscopy (XANES) at the sulfur K-edge to probe the state of Cys221 in the fibrillar form of Ure2pC221 and provide structural information on the structure of Ure2p within fibrils. Although the Ure2p dimer dissociation into its constituent monomers has proven to be a prerequisite for assembly into fibrils, we showed the ability of every Ure2pC221 monomer to establish disulfide bonds upon incubation of the fibrils under oxidizing conditions. Our result indicates either that the constituent unit of the fibrillar form of the protein is a dimeric Ure2p or that the fibrils are made of protofilaments assembled in such a way that the residue C221 from a Ure2p molecule in one protofilament is located in the vicinity of a C221 residue from another molecule belonging to a neighbor protofilament.

Disulfides↗

PubMed2005

Chromium oxidation state imaging in mammalian cells exposed in vitro to soluble or particulate chromate compounds.

Hexavalent chromium compounds are known carcinogens for the respiratory tract in humans. The mechanism of cell transformation by hexavalent chromium compounds is not fully understood although a role for intracellular reduction is sought. The aim of this study was to determine the distribution of Cr valence states in human cells after in vitro exposure to soluble or particulate chromium compounds. A synchrotron X-ray-based microprobe was used to investigate the cellular reduction of Cr(VI) and to image chromium oxidation states in cells. It was shown that soluble Cr(VI) compounds are fully reduced to Cr(III) in cells. Cr(III) is homogeneously distributed within the cell volume and therefore present within the nucleus. In the case of low solubility particulate chromate compounds, Cr(VI) can coexist in the cell environment, as particles in the perinuclear region, together with intracellular and intranuclear Cr(III). Chemical distribution maps also suggest that intracellular Cr(III) originates from extracellular dissolution and reduction of lead chromate rather than from intracellular engulfed particles. The possible stronger carcinogenicity of low solubility chromate vs soluble chromate compounds may derive from the combinative genotoxic effects of intranuclear Cr(III) and the persistent exposure to a strong oxidant, Cr(VI).

Animals↗

PubMed2003

X-ray absorption spectroscopy reveals a substantial increase of sulfur oxidation in transthyretin (TTR) upon fibrillization.

Transthyretin (TTR) amyloid fibrils are the main component of the amyloid deposits occurring in Familial Amyloidotic Polyneuropathy patients. This is 1 of 20 human proteins leading to protein aggregation disorders such as Alzheimer's and Creutzfeldt-Jakob diseases. The structural details concerning the association of the protein molecules are essential for a better understanding of the disease and consequently the design of new strategies for diagnosis and therapeutics. Disulfide bonds are frequently considered essential for the stability of protein aggregates and since in the TTR monomers there is one cysteine residue, it is important to determine unambiguously the redox state of sulfur present in the fibrils. In this work we used x-ray spectroscopy to further characterize TTR amyloid fibrils. The sulfur K-edge absorption spectra for the wild type and some amyloidogenic TTR variants in the soluble and fibrillar forms were analyzed. Whereas in the soluble proteins the thiol group from cysteine (R-SH) and the thioether group from methionine (R-S-CH(3)) are the most abundant forms, in the TTR fibrils there is a significant oxidation of sulfur to the sulfonate form in the cysteine residue and a partial oxidation of sulfur to sulfoxide in the methionine residues. Further interpretation of the data reveals that there are no disulfide bridges in the fibrillar samples and suggest conformational changes in the TTR molecule, namely in strand A and/or in its vicinity, upon fibril formation.

Benzothiazoles↗

PubMed2002

Differential interference contrast x-ray microscopy with twin zone plates.

X-ray imaging in differential interference contrast (DIC) with submicrometer optical resolution was performed by using a twin zone plate (TZP) setup generating focal spots closely spaced within the TZP spatial resolution of 160 nm. Optical path differences introduced by the sample are recorded by a CCD camera in a standard full-field imaging and by an aperture photodiode in a standard scanning transmission x-ray microscope. Applying this x-ray DIC technique, we demonstrate for both the full-field imaging and scanning x-ray microscope methods a drastic increase in image contrast (approximately 20x) for a low-absorbing specimen, similar to the Nomarski DIC method for visible-light microscopy.

Journal Article↗

PubMed2002

Scanning x-ray microscopy of living and freeze-dried blood cells in two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea.

Some ascidians (sea squirts) accumulate the transitional metal vanadium in their blood cells at concentrations of up to 350 mM, about 10(7) times its concentration found in seawater. There are approximately 10 different types of blood cell in ascidians. The identity of the true vanadium-containing blood cell (vanadocyte) is controversial and little is known about the subcellular distribution of vanadium. A scanning x-ray microscope installed at the ID21 beamline of the European Synchroton Radiation Facility to visualize vanadium in ascidian blood cells. Without fixation, freezing or staining realized the visualization of vanadium localized in living signet ring cells and vacuolated amoebocytes of two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea. A combination of transmission and fluorescence images of signet ring cells suggested that in both species the vacuoles contain vanadium.

Animals↗