Search results for "substrate"
showing 10 items of 1018 documents
Affinity chromatography with triazine dyes immobilized onto activated non-porous monodisperse silicas
1988
Abstract Non-porous monodisperse silicas with a particle diameter of 2.1 μm were modified with different silanes for immobilization of various triazine dyes including Procion Red HE3B, Procion Red MX5B, and Cibacron Blue F3GA. Lactate dehydrogenase and malate dehydrogenase from different species and aldehyde reductase from rat brain were purified by affinity elution using the substrate of the enzyme and NADH. With Cibacron F3GA the selectivity for NADH-dependent enzymes was higher than with the two Procion dyes. The utility of these immobilized triazine dye systems on non-porous silica supports for the rapid separation of Cohn Fraction III plasma proteins, including plasminogen, is also des…
Ion-exchange voltammetry at a surfactant-doped electrode: model of mass transfer kinetics to an anionic surface-charged electrode and its application…
2000
cited By 10; International audience; A theoretical model of mass transfer kinetics was developed for investigating the accumulation of cationic compounds at an anionic surfactant-doped screen-printed electrode. It was extended to an enzyme-generated cationic product for the indirect determination of alkaline phosphatase (AP). The model takes into account the analyte depletion, the kinetics of the ion-exchange reaction-diffusion and the kinetics of the enzymatic reaction. The relationship between the ion-exchange voltammetric anodic peak current and the enzyme incubation/accumulation time for a given concentration of AP was established and the validity of the theoretical model was verified e…
Screening of acetylcholinesterase inhibitors by CE after enzymatic reaction at capillary inlet.
2009
In this study the development of a procedure based on capillary electrophoresis after enzymatic reaction at capillary inlet methodology for the screening and in vitro evaluation of the biological activity of acetylcholinesterase (AChE) inhibitors is presented. The progress of the enzymatic reaction of the hydrolysis of acetylthiocholine at pH 8 in the presence of AChE and the inhibitor studied is determined by measuring at 230 nm the peak area of the reaction product thiocholine (TCh). In the method employed the capillary was first filled with 30 mM borate-phosphate buffer (pH 8.0) and subsequently, plugs of: (i) water, (ii) AChE solution, (iii) substrate solution with or without inhibitor,…
Secondary structure conformation of hydroperoxide lyase from green bell pepper, cloned in Yarrowia lipolytica, and its activity in selected media
2008
International audience; Circular dichroism (CD) spectroscopy of secondary structure conformation of the purified green bell pepper hydroperoxide lyase (HPL), cloned in the yeast Yarrowia lipolytica, was investigated. The CD spectra of HPL in iso-octane, obtained at 60 °C, in the presence of the reducing agent dithiothreitol showed dramatic increase in α-helix content. The enzyme conformation remained unchanged over a range of pH values of 5.0–7.0. Using 13-hydroperoxide of linoleic acid (13-HPOD) as substrate, the biocatalysis of HPL in organic solvent media, including chloroform, dichloromethane, hexane, iso-octane, octane and toluene, was investigated. The results indicated an increase in…
Core Histones Are Glutaminyl Substrates for Tissue Transglutaminase
1996
Chicken erythrocyte core histones are glutaminyl substrates in the transglutaminase (TGase) reaction with monodansylcadaverine (DNC) as donor amine. The modification is very fast when compared with that of many native substrates of TGase. Out of the 18 glutamines of the four histones, nine (namely glutamine 95 of H2B; glutamines 5, 19, and 125 of H3; glutamines 27 and 93 of H4; and glutamines 24, 104, and 112 of H2A) are the amine acceptors in free histones. The use of Gln112 of H2A requires a temperature-dependent partial unfolding of the histone, showing that structural determinants are decisive for the glutamine specificity. The structures of H2A and H2B do not appreciably change upon mo…
Magnesium alloys (WE43 and ZE41) characterisation for laser applications
2004
International audience; One of the most important parameters in laser treatment is the quantity of beam energy absorbed by the substrate. Despite its important role played in laser processes, this factor is rarely available for the laser sources wavelengths and at high temperatures reached during such treatments. A series of experiments were carried out in order to characterise, from this point of view, two types of magnesium alloys, WE43 and ZE41, often used in laser applications (cladding, alloying, welding, etc.). The results represent an important step in order to understand Mg-alloys behaviour under laser beam action.
Substrate determinants for cleavage in cis and in trans by the hepatitis C virus NS3 proteinase
1995
Processing of the hepatitis C virus polyprotein is accomplished by a series of cotranslational and posttranslational cleavages mediated by host cell signalases and two virally encoded proteinases. Of these the NS3 proteinase is essential for processing at the NS3/4A, NS4A/4B, NS4B/5A, and NS5A/5B junctions. Processing between NS3 and NS4A occurs in cis, implying an intramolecular reaction mechanism, whereas cleavage at the other sites can also be mediated in trans. Sequence analysis of the amino termini of mature cleavage products and comparisons of amino acid residues around the scissile bonds of various hepatitis C virus isolates identified amino acid residues which might contribute to su…
Strategies to In Vitro Assessment of Major Human CYP Enzyme Activities by Using Liquid Chromatography Tandem Mass Spectrometry
2008
At the early stage of drug discovery, thousands of new chemical entities (NCEs) may be screened before a single candidate can be identified for development. Determining the role of CYP enzymes in the metabolism of a compound and evaluating the effect of NCEs on human CYP activities are key issues in pharmaceutical development as they may explain inter-subject variability, drug-drug interactions, non-linear pharmacokinetics and toxic effects. Reliable methods for determining enzyme activities are needed to characterize an individual CYP enzyme and to obtain a tool for the evaluation of its role in drug metabolism in humans. Different liquid chromatography tandem mass spectrometry methodologi…
EFFECT OF SURFACTANT AND SUBSTRATE TEMPERATURE ON THE GROWTH OF Ag FILMS ON A SAPPHIRE SURFACE
1997
The possibility of modification of the thin film growth on an insulating (sapphire) substrate by using a Ga monolayer as a "surfactant" was studied. We found that the films grown in this way are electrically conducting and can emit photoelectrons at much lower thickness than those deposited on a pure substrate. The surfactant stabilizes the positions of Ag atoms on the substrate surface and inhibits the coalescence of small nuclei into bigger islands, even when the film is annealed to 450 K. This fact may be very important for thin metal film technology.
Synthesis of the aggregation pheromone of the Colorado potato beetle from its degradation product
2015
Incubation of the Colorado potato beetle aggregation pheromone, (S)-1,3-dihydroxy-3,7-dimethyl-6-octen-2-one, with antennal or leg extracts from this beetle gave 6-methyl-5-hepten-2-one as the major product. This ketone was used as a substrate in a stereoselective synthesis of the pheromone. It was attached to the butanediacetal of glycolic acid with good stereoselectivity and the desired isomer was further enriched by purification of the product of this reaction on silica gel.