Azasugar inhibitors as pharmacological chaperones for Krabbe disease

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.     

Towards Treating Correlations in Bose Condensates

 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     

Time delay as a key to apoptosis induction in the p53 network

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     

Note on a conjecture of toft

A conjecture of Toft [17] asserts that any 4-critical graph (or equivalently, every 4-chromatic graph) contains a fully odd subdivision ofK ₄. We show that if a graphG has a degree three nodev such thatG-v is 3-colourable, then eitherG is 3-colourable or it contains a fully oddK ₄. This resolves Toft's conjecture in the special case where a 4-critical graph has a degree three node, which is in turn used to prove the conjecture for line-graphs. The proof is constructive and yields a polynomial algorithm which given a 3-degenerate graph either finds a 3-colouring or exhibits a subgraph that is a fully odd subdivision ofK ₄. (A graph is 3-degenerate if every subgraph has some node of degree at most three.)     

Specific carbon-13 labelling of leucine residues in human growth hormone.

Biosynthetic human growth hormone specifically 13C-labelled in the carbonyl positions of all 26 leucine residues has been obtained by recombinant DNA techniques using 13C-labelled leucine and an E. coli strain that requires leucine. It is shown that, on the whole, the labelling is specific with no significant mislabelling as would have been the case had the 13C-labelled leucine been metabolized.     

Reactions of the nitrate radical with a series of reduced organic sulphur compounds in air

A 480 L evacuable reaction chamber, equipped with FT-IR spectroscopy on-line and ion chromatography off-line, has been used to study the gas phase reaction between the nitrate radical, NO₃, and the reduced organic sulphur compounds CH₃CH₂SH, (CH₃CH₂)₂S, (CH₃CH₂)₂S₂, and CH₃CH₂SCH₃ in air. The products CH₃CH₂SO₃H, SO₂, H₂SO₄, CH₃CHO, and CH₃CH₂ONO₂ were identified and quantified in the reactions of the first three compounds, CH₃CH₂SH, (CH₃CH₂)₂S, and (CH₃CH₂)₂S₂. The reaction products were CH₃CH₂SO₃H, CH₃SO₃H, SO₂, H₂SO₄, CH₃CHO, and CH₂O in the reaction of CH₃CH₂SCH₃. On the basis of identified reaction products and intermediates observed in the infrared spectra, mechanisms are proposed for the reactions between the NO₃ radical and the four reduced organic sulphur compounds. The results of this study, together with those from previous experiments performed in this laboratory on CH₃SCH₃, CH₃SH, and CH₃SSCH₃ lead to the conclusion that all these species, in the reaction with the NO₃ radical, follow a similar degradation mechanism producing SO₂, H₂SO₄, R? SO₃H, R? CHO, and R? CH₂ONO₂, as the main reaction products. The inital step of the reaction of NO₃ with R? S? R and R? S? H type (R = CH₃, CH₂CH₃) reduced organic sulphur compounds was found to be H-atom abstraction, probably after the formation of an initial adduct. For the reaction between NO₃ and R? S? S? R type compounds, evidence for an addition-decomposition reaction, as the initial steps, was obtained. R? S·, R? S(O)·, and R? S(O)₂· appear to be formed as intermediates in all the reactions. © John Wiley & Sons, Inc.     

An adaptive pruning algorithm for the discrete L-curve criterion

We describe a robust and adaptive implementation of the L-curve criterion. The algorithm locates the corner of a discrete L-curve which is a log–log plot of corresponding residual norms and solution norms of regularized solutions from a method with a discrete regularization parameter (such as truncated SVD or regularizing CG iterations). Our algorithm needs no predefined parameters, and in order to capture the global features of the curve in an adaptive fashion, we use a sequence of pruned L-curves that correspond to considering the curves at different scales. We compare our new algorithm to existing algorithms and demonstrate its robustness by numerical examples.