Search results for "Polyacrylamide gel electrophoresis"
showing 10 items of 89 documents
Effects of Citrus Exocortis Viroid Infection on the Peroxidase/IAA-Oxidase System of Gynura aurantiaca and Lycoperszcon esculentum
1987
Summary The peroxidase/indoleacetic acid (IAA) oxidase system of plants of Gynura aurantiaca and Lycopersicon esculentum healthy and infected by citrus exocortis viroid (CEV) was studied. In these hosts, the infection induced an increase of both enzymic activities, when they were measured between 40 and 50 d post inoculation. The analysis of isozyme composition by polyacrylamide gel electrophoresis and specific staining of peroxidase revealed an intensification of most of the zymogram bands as well as the premature appearance of one of the isozymes in the two plants investigated. All the tomato isoperoxidases exhibited IAA-oxidase activity when measured with the oxygen electrode. These resu…
The nucleus negatively controls the synthesis of mitochondrial proteins in the sea urchin egg.
1983
Enucleation of Paracentrotus lividus eggs, followed by parthenogenetic activation induces a sharp increase in the synthesis of mitochondrial proteins as shown by electrofluorography after in vivo labeling with radioactive amino acids. These results further substantiate the hypothesis that the cell nucleus negatively controls mitochondrial replication in the sea urchin egg.
Characterization of EprA, a major extracellular protein of Oenococcus oeni with protease activity
2008
International audience; Extracellular proteins from Oenococcus oeni. a wine-making bacterium, were isolated during growth on media differing by their nitrogen content. Analysis by two-dimensional electrophoresis revealed a low number of protein signals. Among the main spots, one signal corresponded to a single protein, which contained a lysine repeat domain characteristic of cell-wall hydrolases. We demonstrated that this major protein, named EprA, was able to hydrolyse several proteins. The heterologous production of this protein in Escherichia coli confirmed the protease activity of EprA. With a MW of 21.3 kDa and a pl of 5.3, EprA presents optimal activity at pH 7.0 and 45 degrees C. Thi…
Influence of hydrophobic matching on association of model transmembrane fragments containing a minimised glycophorin A dimerisation motif
2005
AbstractThe principles that govern the folding and packing of membrane proteins are still not completely understood. In the present work, we have revisited the glycophorin A (GpA) dimerisation motif that mediates transmembrane (TM) helix association, one of the best-suited models of membrane protein oligomerisation. By using artificial polyleucine TM segments we have demonstrated in this study that a pattern of only five amino acids (GVxxGVxxT) promotes specific dimerisation. Further, we have used this minimised GpA motif to assess the influence of hydrophobic matching on the TM helix packing process in detergent micelles and found that this factor modulates helix–helix association and/or d…
D-Malic enzyme of Pseudomonas fluorescens.
1982
By the enrichment culture technique 14 gram-negative bacteria and two yeast strains were isolated that used D(+)-malic acid as sole carbon source. The bacteria were identified as Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas aeruginosa and Klebsiella aerogenes. In cell-free extracts of P. fluorescens and P. putida the presence of malate dehydrogenase, D-malic enzyme (NAD-dependent) and L-malic enzyme (NADP-dependent) was demonstrated. D-Malic enzyme from P. fluorescens was purified. Stabilization of the enzyme by 50 mM ammonium sulphate an 1 mM EDTA was essential. Preparation of D-malic enzyme that gave one band with disc gel electrophoresis showed a specific activity of 4-5 U/mg…
In vitro studies of adsorption of milk proteins onto tooth enamel
2006
Abstract The aim of this investigation was to study in vitro adsorption of milk proteins onto tooth enamel. In vitro “milk protein pellicles” were formed on enamel specimens incubated in fluid milk products: skimmed milk (pH 6.7), acidified skimmed milk (pH 4.2), yoghurt and neutralized yoghurt (pH 6.7). The enamel specimens were used as such or pre-incubated in saliva. The “milk pellicles” were desorbed from enamel surfaces, and proteins separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were identified by their pattern of migration in SDS-PAGE or by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) fingerp…
Isolation and characterization of a fish F-type lectin from gilt head bream (Sparus aurata) serum.
2007
A novel fucose-binding lectin, designated SauFBP32, was purified by affinity chromatography on fucose-agarose, from the serum of the gilt head bream Sparus aurata. Electrophoretic mobility of the subunit revealed apparent molecular weights of 35 and 30 kDa under reducing and non-reducing conditions, respectively. Size exclusion analysis suggests that the native lectin is a monomer under the selected experimental conditions. Agglutinating activity towards rabbit erythrocytes was not significantly modified by addition of calcium or EDTA; activity was optimal at 37 degrees C, retained partial activity by treatment at 70 degrees C, and was fully inactivated at 90 degrees C. On western blot anal…
Biochemical genetic differentiation between Pomatoschistus marmoratus and P. tortonesei
1999
Several diagnostic genetic markers were identified in Pomatoschistus marmoratus and P. tortonesei using polyacrylamide gel electrophoresis (PAGE) of allozymes. Twenty-one loci were resolved, including the electrophoretic pattern of muscle proteins. The MDH*, PGM1,2*, EST-1,2*, FUM* and PGI-2* loci exhibited different alleles which were fixed for the two species being analysed. Genetic distance, as calculated by Nei's index, showed a value of 0.413. Environmental hypersalinity, could have influenced the geographical distribution of P. tortonesei.
Intracellular compartmentation and regulation of two shikimate dehydrogenase isoenzymes in Pisum sativum
1974
Summary Pea seeds as well as sprouts and roots contain two isoenzymes of shikimate dehydrogenase. Both isoenzymes can be separated by Polyacrylamide gel electrophoresis as well as through ammonium sulfate fractionation. The molecular weight of both isoenzymes are the same although the net electric charge is different. The Km value for isoenzyme 1 is 3,5 × 10 −4 and the Km value for isoenzyme 2 is 1,67 × 10 −4 M. 3,4-dihydroxybenzoic acid, 3,5-dihydroxybenzoic acid, gallic acid, anthranilic acid and p-methoxycinnamic acid inhibited both isoenzymes competitively. Anthranlic acid showed the largest affinity to both isoenzymes. M-methoxycinnamic acid and m-nitrocinnamic acid inhibited both isoe…
Analysis of the polypeptide composition of the cell walls of Neurospora crassa. Similarities with the proteinaceous material secreted by the slime va…
1991
The polypeptide composition of cell walls from the wild-type strain of Neurospora crassa is compared with that of the proteinaceous extracellular coat (PEM) secreted by the slime strain of this fungus. Analyses included determination of the polypeptide pattern by polyacrylamide gel electrophoresis and blotting followed by staining with Concanavalin A and antibodies raised against the overall antigenic components present in either (whole cell walls or PEM) structure. A complex protein assortment was found associated to the walls of the wild type strain. The similarities observed between the polypeptide patterns of the cell walls and PEM, in addition to the immunological cross-reactivity exhi…