Synthesis of glucosylceramide by purified calf spleen beta-glucosidase

Radioactive glucocerebroside was formed when purified calf spleen β-glucosidase was incubated in the presence of 4-methylumbelliferyl-β-$\text{D}$-glucoside and (¹⁴C) labeled ceramide. The product was identified by cochromatography on thin layer plates and carrier dilution and crystallization to constant specific activity. The radioactive glucocerebroside was also a substrate for hydrolysis by this same β-glucosidase preparation employed for its synthesis resulting in the liberation of labeled ceramide. Neither methyl-β-$\text{D}$-glucoside nor free D-glucose were effective in producing radioactive glucocerebroside when incubated with labeled ceramide in this system.     

Evidence for myosin heterogeneity inDrosophila melanogaster

Summary Electrophoresis of myosin extracts from larvae and adult tissues ofDrosophila melanogaster under non-dissociating conditions indicate that two of the bands seen are myosins. They stain for Ca²⁺ ATPase activity and when cut and re-run under dissociating conditions are found to contain a myosin heavy chain that co-migrates with rabbit skeletal muscle myosin heavy chain. One of the forms of myosin seen is found primarily in extracts from the leg. The other is common to the adult fibrillar flight muscles and the larval body wall muscles.The electrophoretic evidence for two myosin types is strengthened by the histochemical demonstration of two myofibrillar ATPases on the basis of their lability to acid or alkali preincubation. The myofibrillar ATPase in the leg and the Tergal Depressor of the Trochanter (TDT) are shown to be relatively acid labile and alkali stable. The larval body wall muscles and the adult fibrillar flight muscles have an ATPase which is acid stable and alkali labile. This distribution of the two myofibrillar ATPase coincides with that predicted by electrophoresis of extracts from whole tissue and also locates the two myosins to specific muscle types.     

Control mechanisms of tanning in Corcyra cephalonica

Localisation of a prophenoloxidase in the cytoplasm of plasmatocytes was histochemically demonstrable in Corcyra cephalonica, by incubating the blood with different phenolic substrates. The activation of this latent enzyme was found to be controlled by a protein factor present in the cuticle. The haemolymph prophenoloxidase could be activated in vitro by prior incubation of the blood sample at 0°C. The treatment with NaCl, EDTA, or detergents did not cause any activation. The exposure of blood samples to gamma-rays, repeated freezing and thawing, or addition of the cuticular activator, brought about rapid disruption of the haemocytes causing a release of phenoloxidase and acceleration of melanization of the blood. The administration of α-ecdysone to the last instar larvae induced premature pupation and accentuated the activation of phenoloxidase without altering the level of enzyme activity. The possible regulatory mechanisms of tanning during the development of Corcyra have been discussed.     

Phosphoenolpyruvate Carboxylase Purified from Leaves of C3, C4, and C3-C4 intermediate species of Alternanthera: Properties at Limiting and Saturating Bicarbonate

Photosynthetica - Phosphoenolpyruvate carboxylase (PEPC) was purified from leaves of four species of Alternanthera differing in their photosynthetic carbon metabolism: Alternanthera sessilis (C3),...     

Characterization of antibodies to chicken riboflavin carrier protein: Antigenicity of the tryptic fragments

The antigenecity of tryptic fragments of reduced and carboxymethylated chicken riboflavin carrier protein were studied. The tryptic sites of the native riboflavin carrier protein bound to riboflavin were inaccessible. The molecular weight and the elution profile on high performance liquid chromatography (TSK 545 DEAE) were unaltered at an enzyme to substrate ratio of 1:31. However, carboxymethylated riboflavin carrier protein could be cleaved into 3 or 4 fragments at an enzyme to substrate ratio of 1:250 or 1:125. Chromatographic separation of the tryptic fragments on high pressure liquid chromatography (TSK 545 DEAE) revealed the presence of two fragments with different elution profiles but similar molecular weight 26 ±2 kDa. Only one fragment (associated with peak 2) had the ability to displace chicken riboflavin carrier protein in an homologous chicken riboflavin carrier protein radioimmunoassay. Thus, carboxymethylated ribotlavin carrier protein which does not compete with chicken riboflavin carrier protein in the radioimmunoassay, on mild trypsinization generates a fragment which interacts with chicken riboflavin carrier protein in radioimmunoassay.     

Clinical, neurophysiological, biochemical and morphological features of eyes in Persian cats with mannosidosis.

The clinical, neurophysiological, morphological and biochemical manifestation of eyes from Persian kittens affected with alpha-mannosidosis were studied. Clinically the disease is characterized by progressive corneal and lenticular opacification. In addition there is asymmetry in shape and latency of signal conductions which were demonstrated by visual evoked potential studies. Morphological and histochemical studies revealed vacuolization of various ocular cell types which stained positively with Concanavalia ensiformis agglutinin (Con A) and wheat germ agglutinin (WGA). Biochemical studies illustrated low activity of acid alpha-mannosidase in cultured keratocytes and abnormal storage of partially degraded oligosaccharides in these cells, in vitreous humor and lens. This comprehensive study of ocular alpha-mannosidosis demonstrates enzyme deficiency which leads to abnormal storage of oligosaccharides in affected cells and is manifested by morphological alterations and functional impairment.