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.     

Molecular orbital momentum distributions and binding energies for nitric oxide

Binding energy spectra of the valence electrons of the open shell molecule NO have been obtained up to 55 eV at azimuthal angles of 0° and 7° using binary (e, 2e) spectroscopy at an impact energy of 1200 eV. The momentum distribution has been obtained for the least tightly bound (unpaired) electron, removal of which leads to formation of the X ¹Σ⁺ ground state of NO⁺. Momentum distributions have also been measured at 21.0 and 40.5 eV. The measured momentum distributions are compared with several literature wavefunctions of varying complexity. They are found to be in excellent agreement with those calculated using the natural spin orbital wavefunctions of Kouba and Ohrn.     

Structural properties of charged diblock copolymer solutions

Aqueous micellar solutions of ionic/neutral block copolymers have been studied by light scattering, small angle neutron scattering and small angle X-ray scattering. We made use of a polymer comprised of a short hydrophobic block (polyethylene-propylene) PEP and of a long polyelectrolytic block (polystyrene-sulfonate) PSSNa which has been shown previously to micellize in water. The apparent polydispersity of these micelles is studied in detail, showing the existence of a few large aggregates coexisting with the population of micelles. Solutions of micelles are found to order above some threshold in polymer concentration. The order is liquid-like, as demonstrated by the evolution with concentration of the peak observed in the structure factor (), and the degree of order is found to be identical over a large range of concentrations (up to 20 wt%). Consistent values of the aggregation number of the micelles are found by independent methods. The effect of salt addition on the order is found to be weak. Received: 19 June 1997 / Received in final form: 4 September 1997 / Accepted: 9 October 1997     

Chvátal closures for mixed integer programming problems

Chvátal introduced the idea of viewing cutting planes as a system for proving that every integral solution of a given set of linear inequalities satisfies another given linear inequality. This viewpoint has proven to be very useful in many studies of combinatorial and integer programming problems. The basic ingredient in these cutting-plane proofs is that for a polyhedronP and integral vectorw, if max(wx|x P, wx integer} =t, thenwx t is valid for all integral vectors inP. We consider the variant of this step where the requirement thatwx be integer may be replaced by the requirement that be integer for some other integral vector . The cutting-plane proofs thus obtained may be seen either as an abstraction of Gomory's mixed integer cutting-plane technique or as a proof version of a simple class of the disjunctive cutting planes studied by Balas and Jeroslow. Our main result is that for a given polyhedronP, the set of vectors that satisfy every cutting plane forP with respect to a specified subset of integer variables is again a polyhedron. This allows us to obtain a finite recursive procedure for generating the mixed integer hull of a polyhedron, analogous to the process of repeatedly taking Chvátal closures in the integer programming case. These results are illustrated with a number of examples from combinatorial optimization. Our work can be seen as a continuation of that of Nemhauser and Wolsey on mixed integer cutting planes.Supported by Sonderforschungsbereich 303 (DFG) and by NSF Grant Number ECS-8611841.Supported by NSF Grant Number ECS-8418392 and Sonderforschungsbereich 303 (DFG), Institut für Ökonometrie und Operations Research, Universität Bonn, FR Germany.