RESUMO
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.
Assuntos
Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa , Proteínas de Bactérias/química , Carboidratos/análise , Glicoproteínas/química , Plasmodium falciparum/química , Proteínas de Protozoários/química , Animais , Proteínas de Bactérias/metabolismo , Cromatografia/métodos , Glicoproteínas/metabolismo , Glicosilação , Plasmodium falciparum/metabolismo , Testes de Precipitina , Proteínas de Protozoários/metabolismoAssuntos
Colo do Útero/enzimologia , Sialiltransferases/isolamento & purificação , Animais , Colo do Útero/metabolismo , Epitélio/enzimologia , Epitélio/metabolismo , Feminino , Glicoproteínas/química , Glicoproteínas/metabolismo , Macaca radiata , Ciclo Menstrual/fisiologia , Ácidos Siálicos/química , Sialiltransferases/químicaAssuntos
Galactose/metabolismo , Glicoproteínas/metabolismo , Plasmodium falciparum/metabolismo , Proteínas de Protozoários/metabolismo , Animais , Antígenos de Protozoários/química , Galactose/análise , Glicoproteínas/química , Glicoproteínas/imunologia , Humanos , Técnicas In Vitro , Malária Falciparum/parasitologia , Peso Molecular , Plasmodium falciparum/imunologia , Proteínas de Protozoários/química , Proteínas de Protozoários/imunologiaRESUMO
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.
Assuntos
Parede Celular/química , Micrococcus/química , Polissacarídeos Bacterianos/química , Sequência de Carboidratos , Cromatografia de Afinidade , Cromatografia em Camada Fina , Meios de Cultura , Dimetil Sulfóxido/química , Dimetilformamida/química , Concentração de Íons de Hidrogênio , Micrococcus/ultraestrutura , Dados de Sequência Molecular , Polissacarídeos Bacterianos/análise , Polissacarídeos Bacterianos/isolamento & purificaçãoRESUMO
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.
Assuntos
Parede Celular/química , Micrococcus/química , Polissacarídeos Bacterianos/química , Sequência de Carboidratos , Cromatografia em Gel , Cromatografia em Papel , Cromatografia em Camada Fina , Cinética , Micrococcus/ultraestrutura , Dados de Sequência Molecular , Oxirredução , Peptidoglicano/metabolismo , Fosforilação , Polissacarídeos Bacterianos/isolamento & purificaçãoAssuntos
Colo do Útero/química , Glicoproteínas/isolamento & purificação , Oligossacarídeos/isolamento & purificação , Animais , Configuração de Carboidratos , Sequência de Carboidratos , Feminino , Glicoproteínas/química , Macaca radiata , Dados de Sequência Molecular , Estrutura Molecular , Oligossacarídeos/química , PronaseAssuntos
Antifúngicos/uso terapêutico , Dermatomicoses/tratamento farmacológico , Imidazóis/uso terapêutico , Adulto , Ensaios Clínicos como Assunto , Método Duplo-Cego , Esquema de Medicação , Feminino , Humanos , Imidazóis/administração & dosagem , Masculino , Pomadas , Distribuição Aleatória , Tinha/tratamento farmacológicoRESUMO
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.
Assuntos
Antifúngicos/administração & dosagem , Dermatomicoses/tratamento farmacológico , Imidazóis/administração & dosagem , Adolescente , Adulto , Idoso , Antifúngicos/uso terapêutico , Criança , Ensaios Clínicos como Assunto , Método Duplo-Cego , Esquema de Medicação , Feminino , Humanos , Imidazóis/uso terapêutico , Masculino , Microsporum , Pessoa de Meia-Idade , Pomadas , Distribuição Aleatória , Tinha/tratamento farmacológicoRESUMO
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.
Assuntos
Ascite/patologia , Neoplasias Mamárias Experimentais/patologia , Tretinoína/farmacologia , Animais , Ascite/metabolismo , Proteínas de Transporte/metabolismo , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Citosol/metabolismo , Feminino , Neoplasias Mamárias Experimentais/metabolismo , Camundongos , Receptores do Ácido RetinoicoRESUMO
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.
Assuntos
Antígenos Glicosídicos Associados a Tumores , Neoplasias da Mama/análise , Glicoproteínas/análise , Proteínas de Neoplasias/análise , Antígenos de Neoplasias/análise , Carcinoma Intraductal não Infiltrante/análise , Linhagem Celular , Membrana Celular/análise , Dissacarídeos/análise , Feminino , Glicosaminoglicanos/análise , Antígenos HLA/análise , Testes de Inibição da Hemaglutinação , Humanos , Lectinas , Metástase Neoplásica , Aglutinina de AmendoimRESUMO
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.
