RESUMEN
Complex-type oligosaccharides were detected in the sarcoplasm of muscle fibres from cat and human biceps using lectins and anticarbohydrate antibodies. The lectin Datura stramonium agglutinin strongly stained type II A fibres as identified by myosin ATPase activity after alkaline and acid preincubation. In contrast, all muscle fibres showed a moderate coarse granular staining after incubation with Tetracarpidum conophorum agglutinin and Telfairia occidentalis agglutinin which recognize tri-antennary complex glycans poorly bound by D. stramonium agglutinin. Strong sarcoplasmic staining in all muscle fibres was obtained after incubation with an antibody against branched N-acetyllactosamine structure while an antibody against binary 2 --> 3 sialyllactosamine glycans failed to detect the muscle fibres. Treatment of the muscle sections with sialidase prior to incubation with D. stramonium agglutinin did not influence the lectin staining pattern. Staining of blots from electrophoretically separated muscle proteins obtained by homogenization, solubilization and centrifugation of small muscle pieces showed D. stramonium agglutinin binding to a number of bands ranging from 200 kDa to 30 kDa. No D. stramonium agglutinin positive bands were observed in blots from separated mitochondrial proteins while blots from sarcoplasmic reticulum separated by electrophoresis stained many bands in the range from 200 kDa to 30 kDa. It may be concluded that all muscle fibres in human and cat biceps hold intracellular non-sialylated complex-type oligosaccharides and further, that a specific tri-antennary complex-type glycoform is strongly expressed in type II A fibres as recognized by D. stramonium agglutinin. These results indicate a different glycosylation of certain myofibrillar-associated proteins in muscle fibre types.
Asunto(s)
Datura stramonium , Lectinas/metabolismo , Fibras Musculares Esqueléticas/química , Plantas Medicinales , Plantas Tóxicas , Animales , Secuencia de Carbohidratos , Gatos , Colorantes , Femenino , Humanos , Persona de Mediana Edad , Datos de Secuencia Molecular , Peso Molecular , Miosinas/análisis , Lectinas de PlantasRESUMEN
The amino-acid sequences of the subunits of the lectin BMA from seeds of Bowringia mildbraedii have been determined. The data indicate that the lectin consists of a precursor polypeptide of approx. 29 kDa that is cleaved almost completely into two fragments of approx. 13.3 kDa (alpha subunit) and approx. 11.9 kDa (beta subunit), respectively. The beta subunit represents the N-terminal half of precursor polypeptides and is disulphide-linked in a beta beta dimer in the native (alpha beta)2 protein. BMA shows extensive amino-acid sequence homologies with known legume lectins. The site of post-translational proteolysis of the putative precursor occurs at a position similar to that identified in lectins obtained from other Sophoreae plants such as Sophora japonica and in Diocleae lectins such as Concanavalin A, but different from that of two chain lectins obtained from other tribes of the Papilionaceae.
Asunto(s)
Aglutininas/química , Fabaceae/química , Plantas Medicinales , Precursores de Proteínas/química , Secuencia de Aminoácidos , Aminoácidos/análisis , Sitios de Unión , Concanavalina A/química , Disulfuros , Datos de Secuencia Molecular , Lectinas de Plantas , Homología de Secuencia de AminoácidoRESUMEN
Affinity chromatography on Bowringia mildbraedii agglutinin (BMA) Sepharose of glycopeptides confirmed a previous report using oligosaccharides (Animashaun, T. and Hughes, R. C. (1989) J. Biol. Chem. 264,4657-4663) that high affinity binding requires the sequence Man alpha 1---2 Man alpha 1----6 Man alpha 1----6 Man beta 1----4. However, moderate binding was still exhibited by structures lacking this sequence provided the oligosaccharide core sequence Man alpha 1----3[Man alpha 1----6]Man beta 1----4GlcNAc was present. This moderate binding was not affected by substitution with N-acetylglucosamine at C2 and C4, respectively, of the Man alpha 1----3 and Man beta 1----4 residues and BMA Sepharose should prove to be a useful tool for the isolation of bisected or non-bisected hybrid-type glycans.
Asunto(s)
Aglutininas/metabolismo , Fabaceae/metabolismo , Plantas Medicinales , Polisacáridos/metabolismo , Aglutininas/química , Secuencia de Carbohidratos , Cromatografía de Afinidad , Glicopéptidos/química , Glicopéptidos/metabolismo , Datos de Secuencia Molecular , Oligosacáridos/química , Oligosacáridos/metabolismo , Polisacáridos/químicaAsunto(s)
Fibroblastos/efectos de los fármacos , Preparaciones de Plantas , Proteínas de Plantas , Ricina/farmacología , Toxinas Biológicas/farmacología , Animales , Línea Celular , Membrana Celular/metabolismo , Cricetinae , Resistencia a Medicamentos , Fibroblastos/metabolismo , Glicoproteínas/metabolismo , Riñón/efectos de los fármacos , Lectinas/farmacología , Hígado/efectos de los fármacos , Ratones , Muérdago/análisis , Lectinas de Plantas , Plantas Medicinales , Biosíntesis de Proteínas , Ratas , Proteínas Inactivadoras de Ribosomas Tipo 2 , Toxinas Biológicas/metabolismoAsunto(s)
Bacillus subtilis/análisis , Pared Celular/análisis , Mutación , Bacillus subtilis/citología , Bacillus subtilis/crecimiento & desarrollo , Cromatografía DEAE-Celulosa , Cromatografía por Intercambio Iónico , Cromatografía en Papel , Medios de Cultivo , Glutamatos/análisis , Hexosaminas/análisis , Mucoproteínas/análisis , Pentosafosfatos/análisis , Fósforo/análisis , Ácidos Pimélicos/análisis , Polisacáridos Bacterianos/análisis , Cloruro de SodioRESUMEN
A double-blind cross-over trial over 24 weeks (10 weeks on the active remedy, 4 weeks off treatment, and 10 weeks on placebo) of the effect of L-dopa on idiopathic Parkinsonism (paralysis agitans) has shown no difference in the response obtained in patients who had undergone previous stereotaxic ventrolateral thalamotomy and in those who had not. Of the 34 patients (18 men and 16 women) in the trial 18 had been operated on (nine unilateral, nine bilateral operations) and 16 had not. All patients entering the trial were taking anticholinergic drugs in stable dosage and these were continued throughout. The only factor which seemed to limit the response to treatment was pre-existing hypertension. Of 31 patients who completed the 10-week treatment period, 12 showed marked improvement, 15 moderate improvement, and 4 and mild or negligible change. It seems that previous ventrolateral thalamotomy affords some protection against the development of L-dopa-induced involuntary limb movements on the side contralateral to the operation. As found by others, maximum benefit was seen in bradykinesia and rigidity and related features but a significant reduction in tremor was also noted during treatment. Side effects (nausea, hypotension, and involuntary movements) were common but rarely limited the therapeutic response.