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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 singular-perturbation expansion of the plasma cold-fluid equations for crossed fields in a planar geometry is presented. The general expansion is carried out to third order. Various instabilities that occur in the first, second, and third orders are discussed. 相似文献
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We use the adiabatic hyperspheric expansion and the Faddeev decomposition of the wave function with only s-waves. We derive for a fixed hyperradius an integro-differential equation for the angular eigenvalue and wave function. The
correlations lower the interaction energy for N = 20 by about a factor of 5.
Received October 22, 2001; accepted for publication November 5, 2001 相似文献
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G. Tiana M.H. Jensen K. Sneppen 《The European Physical Journal B - Condensed Matter and Complex Systems》2002,29(1):135-140
A feedback mechanism that involves the proteins p53 and mdm2, induces cell death as a controlled response to severe DNA damage.
A minimal model for this mechanism demonstrates that the response may be dynamic and connected with the time needed to translate
the mdm2 protein. The response takes place if the dissociation constant k between p53 and mdm2 varies from its normal value. Although it is widely believed that it is an increase in k that triggers the response, we show that the experimental behaviour is better described by a decrease in the dissociation
constant. The response is quite robust upon changes in the parameters of the system, as required by any control mechanism,
except for few weak points, which could be connected with the onset of cancer.
Received 8 May 2002 / Received in final form 9 July 2002 Published online 17 September 2002 相似文献
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Christophe Roussel Loc Dayon Henrik Jensen Hubert H. Girault 《Journal of Electroanalytical Chemistry》2004,570(2):187-199
A series of electrogenerated selective electrophiles based on substituted benzoquinones has been characterized as tags for l-cysteine and cysteine residues in proteins. The electrophiles are generated electrochemically from the corresponding hydroquinones. It is shown from mass spectrometry analysis that the electrogenerated benzoquinone can tag the biomolecules. The rate constants pertaining to the addition of l-cysteine onto the electrogenerated benzoquinones have been determined using electrochemical techniques. The substitution patterns have been unraveled leading to the assessment of site-specific rate constants. It is shown that the rate constants are primarily dependent on the electronic nature of the substituents as expressed by the Hammett substitution constant. The apparent tagging yields observed for l-cysteine in nanospray mass spectrometry experiments do not correspond to the yields expected from the electrochemical study, as the ionisation efficiencies are highly dependent on the tag. Finally, the on-line tagging has been tested using β-lactoglobulin A and myoglobin. Based on these results, it is concluded that the tagging reaction is selective towards cysteine when it takes place in the nanospray interface. The results show that the methodology presented can be used for a rapid characterization and identification of reactive sites in biomolecules. 相似文献
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Hebb J.P. Jensen K.F. Thomas J. 《Semiconductor Manufacturing, IEEE Transactions on》1998,11(4):607-614
The radiative properties of patterned silicon wafers have a major impact on the two critical issues in rapid thermal processing (RTP), namely wafer temperature uniformity and wafer temperature measurement. The surface topography variation of the die area caused by patterning and the roughness of the wafer backside can have a significant effect on the radiative properties, but these effects are not well characterized. We report measurements of room temperature reflectance of a memory die, logic die, and various multilayered wafer backsides. The surface roughness of the die areas and wafer backsides is characterized using atomic force microscopy (AFM). These data are subsequently used to assess the effectiveness of thin film optics in providing approximations for the radiative properties of patterned wafers for RTP applications 相似文献