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The SUMO-conjugating enzyme Ubc9 is an essential enzyme in the SUMO (small ubiquitin-related modifier) protein modification system. Although sumoylation, covalent modification of cellular proteins by SUMO, is considered to regulate various cellular processes, and many substrates for sumoylation have been identified recently, the regulation of Ubc9 expression has not been examined in detail. We analyzed the expression of Ubc9 during rat brain development at the mRNA and protein levels. Northern and Western blot analyses revealed that expression of Ubc9 and SUMO-1 was developmentally regulated, while that of the ubiquitin-conjugating enzyme UbcH7 did not change so dramatically. In situ hybridization analysis revealed that the expression of Ubc9 was high in neuronal stem cells and moderate in differentiated neurons at embryonic stages. In the adult brain, moderate expression was observed in subsets of neurons, such as the dentate granular neurons and pyramidal neurons in the hippocampal formation and the large pyramidal neurons in the cerebral cortex. These results suggest that the Ubc9-SUMO system might participate in the proliferation and differentiation of neuronal cells in the developing brain and in neuronal plasticity in the adult brain.
Proteasomes or multicatalytic proteinases have recently been discovered in mammalian cells. These enzymes are major non-lysosomal proteinases of eukaryotic cells that can account up to 1% of the soluble cellular protein. Our previous studies indicated that multicatalytic proteinases are present in parasitic nematodes of sheep. In this study we have demonstrated that the method described by Rivett et al. (1994) and used for mammalian cells, can give reasonably good preparations of proteasomes from Trichostrongylus colubriformis infective larvae. Further, the inactivation of proteasomes activity by antisera suggests that enzymatic activity of proteasomes is recognised by the immune system of sheep.
Metastasis accounts for most of deaths caused by cancer. The increasing body of evidence suggests that changes in N-glycosylation of tumor cell proteins such as increased branching, increased sialylation, polysialylation, decreased fucosylation, enhanced formation of Lewis X and sialyl Lewis X antigens are among important factors determining metastatic potential of tumor cell. Most of the adhesion proteins, e.g., integrins, members of immunoglobulin superfamily, and cadherins are heavily N-glycosylated. The other proteins involved in adhesion, like galectins and type-C selectins, recognize N-glycans as a part of their specific ligands. In this review we focus on recent reports concerning the contribution of N-glycosylation of tumor cell adhesion molecules and some selected membrane proteins in the tumor invasion and metastasis.
 Phage ΦC31 integrase is a potential vector for the insertion of therapeutic genes into specific sites in the human genome. To understand the mechanism involved in ΦC31 integrase-mediated recombination, it is important to understand the interaction between the integrase and cellular proteins. Using a yeast two-hybrid system with pLexA-ΦC31 integrase as bait, we screened a pB42AD human fetal brain cDNA library for potential interacting cellular proteins. From the 106 independent clones that were screened, 11 potential interacting clones were isolated, of which one encoded C-terminal fragment of Sp100. The interaction between Sp100 and ΦC31 integrase was further confirmed by yeast mating and co-immunoprecipitation assays. The hybridization between a ΦC31 integrase peptide array and an HEK293 cell extract revealed that residues 81RILN84 in the N-terminus of ΦC31 integrase are responsible for the interaction with Sp100. Knocking down endogenous Sp100 with Sp100-specific siRNA increased ΦC31 integrase-mediated recombination but did not impact reporter gene expression. Therefore, endogenous Sp100 may interact with ΦC31 integrase and inhibit the efficiency of ΦC31 integrase-mediated recombination.
One dimensional patterns of proteins from homogenates and four cellular fractions i. e., nuclear, mitochondrial, microsomal, and cytosolic from normal and leukemic lymphocytes were compared. Results obtained revealed that the neoplastic transformation of normal lymphocytes into CLL and ALL ones is associated with the expression of some novel proteins. Electrophoretically-specific nuclear protein of B-CLL lymphocytes with mol. wt of 38/39 kDa was used as immunogen to produce rabbit antiserum. It was observed that obtained antiserum crossreacted with 38/39 kDa antigen of nuclear fractions from CLL and ALL lymphocytes (15 of 16 studied), but not with any of normal ones. It was shown, by Western blot technique, that the expression of 38/39 kDa antigen is correlated with progression of B-CLL disease.
The range of solar radiation reaching the air-water inter-phase, medium wave UV radiation, i.e. UVB 290–320 nm and UVA 320–400 nm, is of the highest biological importance due to its harmful effects. Radiation within this range causes DNA damage (lethal effect) or limits the growth of organisms by inhibiting enzyme synthesis, reducing active transport, or by inducing mutations. The studies were carried out in 2007 based on samples water collected from surface microlayer (SM) (up to 150 μm) and subsurface water (SSW) (25 cm) of pelagic zone of eutrophic lake. The representative collection of bacterial strains was isolated from collected samples. The following are measured: the DNA and cellular protein synthesis activity, respiration activity of the bacteria and activity of hydrolytic enzymes in control cultures, subjected to UVB radiation (applied dose 100 mW cm⁻²) and with and without humic substances (HS) (final concentration 100 mg L⁻¹) playing role of compounds potentially protective from UV radiation. UVB irradiation had the strongest inhibiting impact on production of DNA in bacterial cells (12–23% of that in non-irradiated samples). UVB radiation also inhibits the synthesis of cellular protein (27–43% of that in non-irradiated samples) and bacterial respiration activity (44–48%). UVB radiation had by far the lowest impact on the activity of hydrolytic enzymes. HS may function as a protective agent against UV radiation only in DNA synthesis. No significant differences in response to UVB were found between planktonic and neustonic bacteria.
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