Glucocorticoid-regulated and constitutive trafficking of proteolytically processed cell surface-associated glycoproteins in wild type and variant rat hepatoma cells

Glucocorticoids regulate the trafficking of mouse mammary tumor virus (MMTV) glycoproteins to the cell surface in the rat hepatoma cell line M1.54, but not in the immunoselected sorting variant CR4. To compare the localization of MMTV glycoproteins to another proteolytically processed glycoprotein, both wild type M1.54 cells and variant CR4 cells were transfected with a human insulin receptor (hIR) expression vector, pRSVhIR. The production of cell surface hIR was monitored in dexamethasone-treated and -untreated wild type M1.54 and variant CR4 cells by indirect immunofluorescence, direct plasma membrane immunoprecipitation, and by [125I] insulin binding. In both wild type and variant rat hepatoma cells, hIR were localized at the cell surface in the presence or in the absence of 1 microM dexamethasone. In contrast, the glucocorticoid-regulated trafficking of cell surface MMTV glycoproteins occurred only in wild type M1.54 cells. We conclude that the hIR, which undergoes posttranslational processing reactions similar to MMTV glycoproteins, does not require glucocorticoids to be transported to the plasma membrane and is representative of a subset of cell surface glycoproteins whose trafficking is constitutive in rat hepatoma cells. Thus, MMTV glycoproteins and hIR provide specific cell surface markers to characterize the glucocorticoid-regulated and constitutive sorting pathways.     

The ω-endotoxin protein family displays a hydrophobic motif that might be implicated in toxicity

A computer-based analysis of hydropathy and surface probability of representative members of each class of the Cry family of proteins was performed. A highly conserved hydrophobic motif within the previously described block, D2, is present not only in lepidopteran toxin genes but also in toxins active against diptera and coleoptera. An interesting feature of this hydrophobic motif is the presence of an aspartic residue (highly hydrophilic) in its middle part. Comparison with the amino acid sequence from diphtheria toxin showed that it also contains a hydrophobic motif similar to the one present in the Bacillus thuringiensis toxins. It also contains an aspartic residue in the middle part and some speculations are presented on the function of this specific region with regard to the toxic mechanism of action.     

Alterations of lipid composition in Friend leukemia cell tumors in mice treated with tumor necrosis factor-α

The monocarboxylate (pyruvate) transporter from pea (Pisum sativum) mitochondria was identified by means of a specific monoclonal antibody. The antibody blocked pyruvate-dependent oxaloacetate metabolism without interfering with the metabolism of malate, -ketoglutarate, or glycine. The antibody also blocked the pyruvate/pyruvate exchange reaction of the partially purified transporter reconstituted into phospholipid membranes. Using the specific monoclonal antibody, the transporter was identified on Western blots as a minor 19 kDa protein.     

Growth of Chaetomium cellulolyticum in solid-state fermentation of sugar cane bagasse treated with water and alkali at several liquid/solid ratios

Sugar cane bagasse was water- or alkali-treated at three liquid/solid (L/S) ratios and its digestibility was measured as microbial protein production of Chaetomium cellulolyticum grown on solid-state fermentation columns. The treatments significantly enhanced fungus growth compared to non-treated bagasse, which was used as a control, although the composition of bagasse did not change greatly. Alkali-treated bagasse reached an average protein content of about 7.6% and the lower the L/S ratio, the higher the protein content. L/S ratio did not have an effect in water-treated bagasse. Protein content of water-treated bagasse was also high, approximately 80% of that one of alkali-treated bagasse. Both treatments look promising to enhance sugar cane bagasse potential as an animal feed.Currently at Universidad Francisco de Miranda. Coro, Venezuela.Currently at Ciclo Básico, Facultad de Ingeniería, Universidad del Zulia. Maracaibo, Venezuela.     

Relative toxicity of native Chilean Bacillus thuringiensis strains against Scrobipalpuloides absoluta (Lepidoptera: Gelechiidae)

The larva of Scrobipalpuloides absoluta , a South American moth, is the most devastating insect pest of tomato production in Chile. The potential for using bacterial insecticides was studied analysing the relative toxicity of native Bacillus thuringiensis (BT) isolates belonging to the Chilean collection. The polymerase chain reaction (PCR) technique was used in order to facilitate the prescreening. Mixtures of homologous specific primers to regions within genes encoding CryI , CryIII and CryIV crystal proteins were employed to generate a PCR product profile of each BT isolate. Four isolates were selected and further characterized by means of SDS-PAGE, Western blot and bioassays on fourth-instar S. absoluta larvae. Relative toxicities were evaluated by LD₅₀ determinations. The entomocidal activity of isolate 121e, an autoagglutinating strain, was threefold higher than toxin synthesized by B. thuringiensis var. kurstaki . This native strain was also active against Culex pipiens larvae, although much less than towards S. absoluta .