RESUMEN
Metabolic labelling of Plasmodium falciparum parasites with [3H]GlcN, [3H]Man, [3H]Gal and [3H]ethanolamine, and subsequent purification by SDS-PAGE of the labelled material provided effective labelling of the MSP-1, 195 kDa, and MSP-2, 42-53 kDa, glycoproteins. Reductive beta-elimination of the MSP-2 released from the gel consisted of glycopeptides containing labelled sugars. Processing of the eliminated components and identification of the sugar residues demonstrated the presence of N-acetylglucosaminitol and N-acetylgalactosaminitol amongst other labelled sugars. Reductive beta-elimination with sodium hydroxide-sodium borotritide-borohydride showed the presence of glucosaminitol and alanine in the hydrolysis products. The MSP-2 was retained on solid phase wheat-germ agglutinin and was released from the lectin by treatment with GlcNAc. Upon treatment with O-glycanase the MSP-2 glycoprotein released labelled amino sugar, and derived oligosaccharides on treatment with exoglycosidases released labelled components corresponding to the metabolically incorporated sugars. Labelled Gal was incorporated into the MSP-2 glycoprotein using [3H]UDP-Gal and galactosyltransferase. The galactosylated glycoprotein released labelled Gal upon treatment with beta-galactosidase. The results of the present study suggest that the carbohydrate chains of the MSP-2 glycoprotein are attached to the protein backbone via GlcNAc- and GalNAc-serine/threonine in O-glycosyl linkage and the glycoprotein has terminal GlcNAc and Gal residues. The carbohydrate moieties of MSP-2, glycoprotein consist mainly of short chains linked to the protein core.
Asunto(s)
Antígenos Bacterianos , Proteínas de la Membrana Bacteriana Externa , Proteínas Bacterianas/química , Carbohidratos/análisis , Glicoproteínas/química , Plasmodium falciparum/química , Proteínas Protozoarias/química , Animales , Proteínas Bacterianas/metabolismo , Cromatografía/métodos , Glicoproteínas/metabolismo , Glicosilación , Plasmodium falciparum/metabolismo , Pruebas de Precipitina , Proteínas Protozoarias/metabolismoAsunto(s)
Cuello del Útero/enzimología , Sialiltransferasas/aislamiento & purificación , Animales , Cuello del Útero/metabolismo , Epitelio/enzimología , Epitelio/metabolismo , Femenino , Glicoproteínas/química , Glicoproteínas/metabolismo , Macaca radiata , Ciclo Menstrual/fisiología , Ácidos Siálicos/química , Sialiltransferasas/químicaAsunto(s)
Galactosa/metabolismo , Glicoproteínas/metabolismo , Plasmodium falciparum/metabolismo , Proteínas Protozoarias/metabolismo , Animales , Antígenos de Protozoos/química , Galactosa/análisis , Glicoproteínas/química , Glicoproteínas/inmunología , Humanos , Técnicas In Vitro , Malaria Falciparum/parasitología , Peso Molecular , Plasmodium falciparum/inmunología , Proteínas Protozoarias/química , Proteínas Protozoarias/inmunologíaRESUMEN
A fragment of Micrococcus lysodeikticus cell wall was obtained by extraction of walls with water, dimethylformamide or dimethyl sulfoxide. The water-soluble polymer was obtained from the cell walls prepared either with or without trypsin treatment of the cell. This fragment was studied by the Smith periodate oxidation, methylation, mild acid treatment and enzymic procedures. The polymer consists of polysaccharide chains composed of (1-->4)-O-(2-acetamido-2-deoxy-beta-D-mannopyranosyluronic acid)-(1-->6)-O-alpha-D-glucopyranosyl residues. The polysaccharide chain is linked to C-6 of a 2-acetamido-3-O-(D-1-carboxyethyl)-2-deoxy-beta-D-gluco-pyranosyl residue of a peptidoglycan chain composed of repeating (1-->4)-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-(1-->4)-[2-acet ami do-3-O-(D-1-carboxyethyl)-2-deoxy-beta-D-glucopyranosyl] residues. The water-soluble cell-wall fragment was also observed in the-culture medium of Micrococcus lysodeikticus and was also extractable from the cells in minor quantity.
Asunto(s)
Pared Celular/química , Micrococcus/química , Polisacáridos Bacterianos/química , Secuencia de Carbohidratos , Cromatografía de Afinidad , Cromatografía en Capa Delgada , Medios de Cultivo , Dimetilsulfóxido/química , Dimetilformamida/química , Concentración de Iones de Hidrógeno , Micrococcus/ultraestructura , Datos de Secuencia Molecular , Polisacáridos Bacterianos/análisis , Polisacáridos Bacterianos/aislamiento & purificaciónRESUMEN
A polysaccharide-peptidoglycan complex containing different phosphorylated sugars from Micrococcus lysodeikticus cell wall has been isolated and purified. The peptidoglycan contained muramic acid 6-phosphate and N-acetylglucosamine 6-phosphate as phosphorylated sugars in addition to other sugar residues. Mild acid hydrolysis of the peptidoglycan and subsequent reduction of the released polysaccharide showed therein the presence of glucose and N-acetyl-glucosamine in the linkage of the external polysaccharide residues to the peptidoglycan through phosphodiester linkage. These data suggest the presence of polysaccharide chains linked to a peptidoglycan core through two phosphorylated sugars via two different terminal carbohydrate residues of the external polysaccharide chains in a same polymer.
Asunto(s)
Pared Celular/química , Micrococcus/química , Polisacáridos Bacterianos/química , Secuencia de Carbohidratos , Cromatografía en Gel , Cromatografía en Papel , Cromatografía en Capa Delgada , Cinética , Micrococcus/ultraestructura , Datos de Secuencia Molecular , Oxidación-Reducción , Peptidoglicano/metabolismo , Fosforilación , Polisacáridos Bacterianos/aislamiento & purificaciónAsunto(s)
Cuello del Útero/química , Glicoproteínas/aislamiento & purificación , Oligosacáridos/aislamiento & purificación , Animales , Conformación de Carbohidratos , Secuencia de Carbohidratos , Femenino , Glicoproteínas/química , Macaca radiata , Datos de Secuencia Molecular , Estructura Molecular , Oligosacáridos/química , PronasaAsunto(s)
Antifúngicos/uso terapéutico , Dermatomicosis/tratamiento farmacológico , Imidazoles/uso terapéutico , Adulto , Ensayos Clínicos como Asunto , Método Doble Ciego , Esquema de Medicación , Femenino , Humanos , Imidazoles/administración & dosificación , Masculino , Pomadas , Distribución Aleatoria , Tiña/tratamiento farmacológicoRESUMEN
A single daily application of oxiconazole cream was shown, in a double-blind, randomized, multicentric study, to be effective in treating various dermatomycoses as well as erythrasma. Tolerance was excellent and side effects were negligible.
Asunto(s)
Antifúngicos/administración & dosificación , Dermatomicosis/tratamiento farmacológico , Imidazoles/administración & dosificación , Adolescente , Adulto , Anciano , Antifúngicos/uso terapéutico , Niño , Ensayos Clínicos como Asunto , Método Doble Ciego , Esquema de Medicación , Femenino , Humanos , Imidazoles/uso terapéutico , Masculino , Microsporum , Persona de Mediana Edad , Pomadas , Distribución Aleatoria , Tiña/tratamiento farmacológicoRESUMEN
Retinoic acid caused a decrease in adhesiveness but no growth change in the allotransplantable TA3-Ha cell and no change in adhesiveness or growth in the strain specific TA3-St cell. The retinoic acid binding protein was detected in the TA3-Ha, but not the TA3-St, cell.
Asunto(s)
Ascitis/patología , Neoplasias Mamarias Experimentales/patología , Tretinoina/farmacología , Animales , Ascitis/metabolismo , Proteínas Portadoras/metabolismo , Adhesión Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Línea Celular , Citosol/metabolismo , Femenino , Neoplasias Mamarias Experimentales/metabolismo , Ratones , Receptores de Ácido RetinoicoRESUMEN
Neuraminidase (Vibrio cholerae) treatment of human metastatic mammary carcinoma MDA-MB-231 cells grown in culture released 0.60-0.63 mg of N-acetylneuraminic acid from 10(9) cells. Incubation of intact cells with a modified trypsin and fractionation by gel filtration gave mainly O-glycopeptides. The presence of O-glycosyl-linked chains having one or two carbohydrate residues was confirmed by treatment of the glycopeptide fractions with galactose oxidase, followed by reduction with alkaline sodium borotritide and fractionation. The major glycopeptide fraction, which consisted of 53% carbohydrate and 47% protein, and a minor glycopeptide fraction each inhibited hemagglutination by peanut lectin. These results suggest the presence of O-beta-D-galactopyranosyl-(1 leads to 3)-O-(2-acetamido-2-deoxy-alpha-D-galactopyranosyl)-(1 leads to 3)-L-serine (threonine) residues. The absorptive capacities for anti-HLA-A2 and anti-HLA-B8 antisera were slightly greater for intact than for lyophilized cells, which suggested that masking of these major histocompatibility antigens did not occur in intact cells.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores , Neoplasias de la Mama/análisis , Glicoproteínas/análisis , Proteínas de Neoplasias/análisis , Antígenos de Neoplasias/análisis , Carcinoma Intraductal no Infiltrante/análisis , Línea Celular , Membrana Celular/análisis , Disacáridos/análisis , Femenino , Glicosaminoglicanos/análisis , Antígenos HLA/análisis , Pruebas de Inhibición de Hemaglutinación , Humanos , Lectinas , Metástasis de la Neoplasia , Aglutinina de ManiRESUMEN
1H-N.m.r.spectra for solutions in dimethyl sulphoxide-d6 of disaccharides related to hyaluronate and chondroitin sulphate are compared with those of their methylated derivatives. All resonances, including those of HO and HN groups, have been assigned. The temperature and concentration dependences suggest that HO-4 of the hexosamine residue in hyalobiouronate (but not that in chondrosinate) is hydrogen-bonded to O-5 of the uronic acid residue. The resonance of HO-2 of the uronate residue of chondrosinate also shows anomalies that may arise from intra-residue hydrogen-bonding. These findings confirm the existence of some features previously suggested to be present in glycosaminoglycuronan polymers. The resonance of HO-4 of the uronate residue in the disaccharides and in sodium (methyl alpha-D-glucopyranosid)uronate behaves as though there was a hydrogen bond between the carboxylate group and HO-4.