Sphingolipid ceramide N-deacylase (SCDase, EC 3.5.1.69) is a hydrolytic enzyme isolated from Pseudomonas sp. TK 4. In addition to its primary deacylation function, this enzyme is able to reacylate lyso-sphingolipids under specific conditions. We immobilised this enzyme on magnetic macroporous cellulose and used it to semisynthesise C17:0 glucosylceramide and C17:0 sulphatide, which are required internal standards for quantification of the corresponding glycosphingolipids (GSL) by tandem mass spectrometry. A high rate of conversion was achieved for both lipids (80% for C17:0 sulphatide and 90% for C17:0 glucosylceramide). In contrast to synthesis with a soluble form of the enzyme, use of immobilised SCDase significantly reduced the contamination of the sphingolipid products with other isoforms, so further purification was not necessary. Our method can be effectively used for the simple preparation of specifically labelled sphingolipids of high isoform purity for application in mass spectrometry.
- MeSH
- amidohydrolasy chemie MeSH
- bakteriální proteiny chemie MeSH
- enzymy imobilizované chemie MeSH
- glukosylceramidy chemická syntéza chemie MeSH
- glykosfingolipidy analýza MeSH
- hmotnostní spektrometrie metody normy MeSH
- hydrolýza MeSH
- Pseudomonas enzymologie MeSH
- referenční standardy MeSH
- stereoizomerie MeSH
- sulfoglykosfingolipidy chemická syntéza chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- amidohydrolasy MeSH
- bakteriální proteiny MeSH
- enzymy imobilizované MeSH
- glukosylceramidy MeSH
- glykosfingolipidy MeSH
- sphingolipid ceramide N-deacylase MeSH Prohlížeč
- sulfoglykosfingolipidy MeSH
In order to obtain an economical, efficient and selective system for glycoprotein modification we prepared reactors with immobilized neuraminidase or (and) galactose oxidase. High storage and operational stability of the enzyme reactors was obtained by their immobilization through the carbohydrate parts of the enzyme molecules to hydrazide-modified supports. Magnetic and non-magnetic forms of bead cellulose and poly(HEMA-co-EDMA) microspheres were used for immobilization. These reactors can be used almost universally for the activation of ligands and for labelling of substances having a carbohydrate moiety.
- MeSH
- enzymy imobilizované chemie metabolismus MeSH
- glykoproteiny chemie metabolismus MeSH
- magnetismus MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- enzymy imobilizované MeSH
- glykoproteiny MeSH
Magnetic hydrogel microspheres 1.5 microm in size were prepared by dispersion copolymerization of 2-hydroxyethyl methacrylate and ethylene dimethacrylate in the presence of magnetite, which formed the core of the particles. RNase A was coupled to the particles by the cyanuric chloride method. Gel electrophoresis of plasmid DNA pUC 19 (contaminated by bacterial RNA) confirmed RNA degradation with the immobilized enzyme. The effect of temperature and pH on the relative activity of immobilized RNase A was estimated after incubation of the samples at different temperatures (30-80 degrees C) and pH (4.0-8.0). Maximum relative activity was observed at 70 degrees C and pH 6.5. The matrices based on magnetic poly(HEMA) had a low tendency to adsorb RNA.
- MeSH
- bakteriální RNA metabolismus MeSH
- biotechnologie metody MeSH
- enzymy imobilizované chemie metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- magnetismus MeSH
- methakryláty chemie MeSH
- mikrosféry * MeSH
- pankreatická ribonukleasa chemie metabolismus MeSH
- polyhydroxyethylmethakrylát chemie MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bakteriální RNA MeSH
- enzymy imobilizované MeSH
- ethylene dimethacrylate MeSH Prohlížeč
- methakryláty MeSH
- pankreatická ribonukleasa MeSH
- polyhydroxyethylmethakrylát MeSH
Immunosorbents for the plant hormones cytokinins prepared by random antibody immobilization (to Affi-Gel 10) and by oriented approach via oxidized carbohydrate moieties on the Fc region (to Affi-Gel Hz or hydrazide derivative of Perloza MT 200) have been compared. Both approaches yielded immunosorbents with high dynamic capacity (ca. 5-10 nmol ml gel-1). Oriented antibody immobilization did not exhibit crucial effects in the case of low-molecular-mass cytokinins. Antibodies immobilized via a spacer to Affi-Gel 10 have probably enough conformational freedom to enable good accessibility to cytokinins. The sorbents were used in analysis of endogenous cytokinins in maize seeds. In phosphatase treated samples trans-zeatin and its riboside were predominant.
- MeSH
- celulosa MeSH
- chromatografie afinitní MeSH
- cytokininy chemie MeSH
- imunochemie MeSH
- imunoglobulin G chemie MeSH
- molekulární konformace MeSH
- oxidace-redukce MeSH
- protilátky chemie MeSH
- semena rostlinná chemie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- celulosa MeSH
- cytokininy MeSH
- imunoglobulin G MeSH
- protilátky MeSH
Immobilization of affinity ligands, proteins, enzymes and other functional groups by azo coupling is based on the high reactivity of the support-carrying diazonium groups towards both low- and high-molecular weight compounds containing certain groupings such as phenols, imidazole and some other heterocycles, thiols and amines. The precursor of diazonium group is a diazotizable amine on the matrix. Remarkable progress in its preparation was achieved by application of (4-amino-phenyl)-(2-sulphatoxyethyl) sulphone-type reagents for functionalization of the matrix. A similar type of amine precursor is prepared by the reaction of monosubstituted sulphanilamide with epoxide-containing matrix. The presence of a SO2 group in the para position to diazonium group of these supports (after diazotization) enhances reactivity in azo coupling. 'Reversed' azo coupling is the reaction of a matrix containing functional groups capable of reacting with diazonium groups of a ligand. Preparation of a suitable matrix and examples of diazotizable ligands are given.
- MeSH
- chromatografie afinitní * MeSH
- diazoniové sloučeniny chemie MeSH
- ligandy MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- diazoniové sloučeniny MeSH
- ligandy MeSH
In 146 individuals with inflammatory periodontal disease the authors verified the therapeutical effect of Sanchelin (the mixture of sanguinarine and chelerythrine in 0.05% concentration) in gel (4% hydrogel carboxymethylcellulose) applied into pockets and in aqueous solution administered in the intrapapillar route. The effect of Sanchelin in solution was compared with the effect of 2% hydrocortison in the same way of administration. The clinical condition of periodontium and the quality of oral hygiene was evaluated by means of indexes and papillae were examined histologically. The authors demonstrated that Sanchelin in gel is an effective drug in treatment of inflammatory periodontal disease. They observed lesser effectivity when intrapapillar administration of Sanchelin in solution was used.
- MeSH
- antiflogistika terapeutické užití MeSH
- gingivitida farmakoterapie MeSH
- lidé MeSH
- parodontitida farmakoterapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiflogistika MeSH
- MeSH
- dějiny 20. století MeSH
- lidé MeSH
- poskytování zdravotní péče * dějiny MeSH
- služby zdravotní péče o dítě MeSH
- studium lékařství dějiny MeSH
- Check Tag
- dějiny 20. století MeSH
- lidé MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- historické články MeSH
- Geografické názvy
- Kuba MeSH
- MeSH
- alkaloidy farmakologie terapeutické užití MeSH
- antiflogistika MeSH
- Bacteria účinky léků MeSH
- benzofenantridiny MeSH
- cholinesterasové inhibitory MeSH
- edém farmakoterapie MeSH
- fenantridiny farmakologie terapeutické užití MeSH
- inbrední kmeny potkanů MeSH
- krysa rodu Rattus MeSH
- mikrobiální testy citlivosti MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- srovnávací studie MeSH
- Názvy látek
- alkaloidy MeSH
- antiflogistika MeSH
- benzofenantridiny MeSH
- cholinesterasové inhibitory MeSH
- fagaronine MeSH Prohlížeč
- fenantridiny MeSH
- MeSH
- alkaloidy terapeutické užití MeSH
- fixní kombinace léků MeSH
- gely MeSH
- hodnocení léčiv MeSH
- isochinoliny terapeutické užití MeSH
- léčivé rostliny * MeSH
- lidé MeSH
- nemoci parodontu farmakoterapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- alkaloidy MeSH
- fixní kombinace léků MeSH
- gely MeSH
- isochinoliny MeSH
