Functional and structural characterization of ovine ornithine transcarbamoylase

Ornithine transcarbamoylase from ovine liver has been purified to homogeneity. Like all anabolic OTCs, the ovine enzyme is a trimer, constituted by identical subunits of 34 kDa. Sequence analysis of the 54 N-terminal residues of ovine OTC shows a high degree of homology with the human enzyme. The optimum pH and the Michaelis constants for the catalytic reaction were determined. The ovine enzyme is the most thermostable one among mammals OTCs, its critical temperature being 6 degrees C higher than those measured for the other enzymes. The enzyme has been crystallised and the structure determined at 3.5 A resolution. Crystals belong to the cubic P4(3)32 space group, with a = b = c = 184.7 A and a solvent content of about 80%. There is no evidence of any ligand in the active site cavity, indicating that the crystals contain an unliganded or T state of the enzyme. The unliganded OTCase enzyme adopts a trimeric structure which, in the crystal, presents a three-fold axis coincident with the crystallographic one. The conformation of each monomer in the trimer is quite similar to that of the liganded human protein, with the exception of a few loops, directly interacting with the substrate(s), which are able to induce a rearrangement of the quaternary organisation of the trimer, that accounts for the cooperative behaviour of the enzyme following the binding of the substrates.     

Terminal Deoxy Hydroxyamino Sugars

Abstract

Reduction of sugar aldoximes gave in good yield the corresponding terminal deoxy hydroxyamino sugars. These compounds were found to be reasonably stable (they could be kept for some weeks at 4° C). On standing in the air, these compounds in solution were spontaneously oxidized to the corresponding nitroxide free radicals whose ESR spectra gave useful structural information.     

Nematic liquid crystal droplets with an enhanced quasi-linear electro-optical response

Nematic liquid crystal droplets dispersed in a thermoplastic matrix with a built-in d.c. electric field exhibit a quasi-linear response to an electric field. In this work we show a device characterized by a large light modulation. The device can store fields up to several V μm^-1 and operates well from d.c. to several kHz. In addition, we found that the experimental results are in agreement with a simple theoretical model for light scattering by a dispersion of liquid crystal droplets. This device allows us to overcome possible drawbacks, due to a reduced light modulation, in applications where polarity detection is required.     

Benzoyl Group Migrations in Methyl 2,3,6-Trio-O-Benzoyl-4-deoxy-4-(N-Hydroxyamino)-α-D-Gluco and α-D-Galacto-Pyranosides

Abstract

Deoxy-N-hydroxyaminosugars constitute a novel family of sugars. The interest in them is mainly due to their close structural similarity to natural sugars, coupled, with their easy oxidation to nitroxide free radicals, enabling their study by ESR spectroscopy.¹ From the point of view of their chemical reactivity, the hydroxyamino group is an ambident nucleophile possessing complex orbital and topographical factors, which direct its reactivity to either of its heteroatoms.     

High Surface Area Mesoporous Silica Nanoparticles with Tunable Size in the Sub-Micrometer Regime: Insights on the Size and Porosity Control Mechanisms

Mesoporous silica nanostructures (MSNs) attract high interest due to their unique and tunable physical chemical features, including high specific surface area and large pore volume, that hold a great potential in a variety of fields, i.e., adsorption, catalysis, and biomedicine. An essential feature for biomedical application of MSNs is limiting MSN size in the sub-micrometer regime to control uptake and cell viability. However, careful size tuning in such a regime remains still challenging. We aim to tackling this issue by developing two synthetic procedures for MSN size modulation, performed in homogenous aqueous/ethanol solution or two-phase aqueous/ethyl acetate system. Both approaches make use of tetraethyl orthosilicate as precursor, in the presence of cetyltrimethylammonium bromide, as structure-directing agent, and NaOH, as base-catalyst. NaOH catalyzed syntheses usually require high temperature (>80 °C) and large reaction medium volume to trigger MSN formation and limit aggregation. Here, a successful modulation of MSNs size from 40 up to 150 nm is demonstrated to be achieved by purposely balancing synthesis conditions, being able, in addition, to keep reaction temperature not higher than 50 °C (30 °C and 50 °C, respectively) and reaction mixture volume low. Through a comprehensive and in-depth systematic morphological and structural investigation, the mechanism and kinetics that sustain the control of MSNs size in such low dimensional regime are defined, highlighting that modulation of size and pores of the structures are mainly mediated by base concentration, reaction time and temperature and ageing, for the homogenous phase approach, and by temperature for the two-phase synthesis. Finally, an in vitro study is performed on bEnd.3 cells to investigate on the cytotoxicity of the MNSs.     

Boundary value problems with nonhomogeneous Neumann conditions on a fractal boundary

This note deals with some boundary value problems in a self-similar ramified domain of ${\mathbb{R}}^2$, with a fractal boundary. The partial differential equation is Laplace's equation, and there are nonhomogeneous generalized Neumann boundary conditions on the fractal boundary. We propose a multiscale strategy for approximating the restriction of the solutions to simple subdomains. This strategy is based on transparent boundary conditions and on a wavelet expansion of the Neumann datum. To cite this article: Y. Achdou, N. Tchou, C. R. Acad. Sci. Paris, Ser. I 342 (2006).     

Study of conformational properties of a biologically active peptide of fibronectin by circular dichroism, NMR and molecular dynamics simulation

Circular dichroism (CD), and NMR spectra have been recorded and molecular dynamics (MD) simulations have been performed in water and water-trifluoroethanol (TFE) mixed solvent for a synthetic biologically active 13-amino-acid fragment of human fibronectin and two related peptides. The CD results are interpreted on the basis of statistical analyses of MD trajectories and of ensuing calculations of CD spectra based on Schellman's matrix method. It is observed that the peptide conformation is quite variable in water and loses its mobility with the addition of TFE. (1)H-NOE data were found to be consistent with the most abundant calculated conformation.     

Alpha-cyclodextrin functionalized CdS nanocrystals for fabrication of 2/3 D assemblies

Different types of cyclodextrins (CDs) have been tested as mediators for the water phase transfer of organic-capped CdS nanocrystals (NCs), and alphaCD has been demonstrated to be the most effective system. The formation of a complex based on alphaCDs and colloidal NCs has been considered to be responsible for the phase transfer process and extensively investigated by optical, structural, and calorimetric measurements, as a function of the experimental parameters (pH and NC and CD concentration). A mechanism for the complexation phenomena has been suggested. The fabrication of 2/3 D supramolecular architectures has been proposed according to two different strategies. First, a layer-by-layer procedure has been used to obtain multilayered structures where polyelectrolyte layers have been intercalated with negatively charged alphaCD-CdS NC complexes by exploiting electrostatic interaction between polyelectrolyte and cyclodextrin OH groups. Second, a monolayer of CdS NCs has been deposited onto a self-assembled monolayer of sulfated CDs, thus combining the use of an electrostatic-force-based approach and host-guest chemistry. The important role played by host-guest interactions has then been revealed.     

α-chymotrypsin superactivity in cetyltrialkylammonium bromide-rich media

α-Chymotrypsin (α-CT) activity was tested with N-glutaryl-l-phenylalanine p-nitroanilide in buffered media with added cationic surfactants. The effect of the commercial cetyltrimethylammonium bromide (CTABr) was compared with that of three other surfactants with ethyl (CTEABr), propyl (CTPABr), and butyl (CTBABr) head groups. These were synthesized and purified in this laboratory. Surfactant head groups provided distinct environments that largely modulated the catalytic performance. Larger alkyl head group hydrophobicity led to a marked enhancement of α-CT activity. CTBABr-rich media induced the highest superactivity. Kinetic measurements were performed in Tris-HCl buffer at a surfactant concentration either below or above CMC, and α-CT superactivity occurred in both media. Positive interactions between the enzyme and surfactants happened independently of thesupramolecular organization of the medium. The reaction followed the Michaelis-Menten kinetics. The substrate to micelle aggregates binding constant was used to calculate the substrate concentration available for catalysis. The k _cat to k _m ratio was in CTBABr-rich media always higher than in pure buffer and depended on the surfactant concentration. α-CT superactivity depended on the pH value of buffer solution. Enzyme inactivation followed a single-step mechanism in pure buffer and a series mechanism in the presence of a surfactant. The rate of activity decay obeyed a first-order kinetics.