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Chris H. Hill Agnete H. Viuff Samantha J. Spratley Stéphane Salamone Stig H. Christensen Randy J. Read Nigel W. Moriarty Henrik H. Jensen Janet E. Deane 《Chemical science》2015,6(5):3075-3086
Krabbe disease is a devastating neurodegenerative disorder characterized by rapid demyelination of nerve fibers. This disease is caused by defects in the lysosomal enzyme β-galactocerebrosidase (GALC), which hydrolyzes the terminal galactose from glycosphingolipids. These lipids are essential components of eukaryotic cell membranes: substrates of GALC include galactocerebroside, the primary lipid component of myelin, and psychosine, a cytotoxic metabolite. Mutations of GALC that cause misfolding of the protein may be responsive to pharmacological chaperone therapy (PCT), whereby small molecules are used to stabilize these mutant proteins, thus correcting trafficking defects and increasing residual catabolic activity in cells. Here we describe a new approach for the synthesis of galacto-configured azasugars and the characterization of their interaction with GALC using biophysical, biochemical and crystallographic methods. We identify that the global stabilization of GALC conferred by azasugar derivatives, measured by fluorescence-based thermal shift assays, is directly related to their binding affinity, measured by enzyme inhibition. X-ray crystal structures of these molecules bound in the GALC active site reveal which residues participate in stabilizing interactions, show how potency is achieved and illustrate the penalties of aza/iminosugar ring distortion. The structure–activity relationships described here identify the key physical properties required of pharmacological chaperones for Krabbe disease and highlight the potential of azasugars as stabilizing agents for future enzyme replacement therapies. This work lays the foundation for new drug-based treatments of Krabbe disease. 相似文献
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The interaction of isotactic polypropylene with ethylene propylene diene terpolymer in their blends has been investigated by use of differential scanning calorimetry, dynamic mechanical analysis, wide- and small-angle x-ray scattering, and by investigating the nucleation and kinetics of crystallization of the iPP component under the polarization microscope. It is found, that the dispersion of the EPDM component in the iPP matrix is dependent on blend composition and is maximal at 10% EPDM content. An interface layer between the two components is formed by migration of iPP molecules into the EPDM phase. A model for this interface is proposed. 相似文献
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Alloys of the systems Fe–Al (mixable over the whole concentration range) and Fe–Mg (insoluble with each other) were produced by implantation of Fe ions into Al and Mg, respectively. The implantation energy was 200 keV and the ion doses ranged from 1 × 1014 to 9 × 1017cm-2The obtained implantation profiles were determined by Auger electron spectroscopy depth profiling. Maximum iron concentrations reached were up to 60 at.% for implantation into Al and 94 at.% for implantation into Mg. Phase analysis of the implanted layers was performed by conversion electron Mössbauer spectroscopy and X‐ray diffraction. For implantation into Mg, two different kinds of Mössbauer spectra were obtained: at low doses paramagnetic doublets indicating at least two different iron sites and at high doses a dominant ferromagnetic six‐line‐pattern with a small paramagnetic fraction. The X‐ray diffraction pattern concluded that in the latter case a dilated αiron lattice is formed. For implantation into Al, the Mössbauer spectra were doublet structures very similar to those obtained at amorphous Fe–Al alloys produced by rapid quenching methods. They also indicated at least two different main iron environments. For the highest implanted sample a ferromagnetic six‐line‐pattern with magnetic field values close to those of Fe3Al appeared. 相似文献
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To investigate local ordering and segregation phenomenon in a Ni91Pt9-alloy after sputtering and annealing a 3D optical atom probe (OAP) has been used. The specimen tips have been prepared from polycrystalline samples. To sputter the samples a separate preparation chamber with a scannable Ar-sputter-gun is connected to the OAP vessel. When necessary, the sample can be electrically heated to induce segregation and cure the altered layer. After a heat treatment of a Ni91 at. %Pt 9 at.% specimen at 1100 K the surface of a (111)-oriented specimen is enriched in platinum by a factor of two in relation to the bulk. The phenomenon of short-range ordering has been investigated on the surface and in the subsurface volume. A 3D reconstruction of this annealed NiPt specimen shows regions with high concentration of platinum that gives an indication at short-range ordering. Uniform sputtering of the tip without a heat treatment induces a decisive depletion of Pt on the surface and the following subatomic layers. The atom-probe results of specimens in thermal equilibrium are in close agreement to further surface sensitive results obtained from Ion Scattering Spectroscopy (ISS) and Auger Electron Spectroscopy (AES). 相似文献