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41.
Gabriele Mezzetti Mariastella Moruzzi Maria G. Monti Giorgio Piccinini Bruno Barbiroli 《Molecular and cellular biochemistry》1985,66(2):175-183
Summary A cyclic nucleotide-independent protein kinase which phoshorylates preferentially acidic proteins such as casein or phosvitin was isolated from cytosol of chick duodenal mucosa. The enzyme was purified more than 633 fold to apparent homogeneity by ammonium sulfate fractionation, column chromatography on DEAE-cellulose, phosphocellulose, hydroxylapatite and by sucrose density gradient centrifugation. The native enzyme has a molecular weight of 131000 as measured by gel filtration. The enzyme is a complex protein containing three polypeptides of molecular weight of 39 000, 36 000 and 27 000. It behaves as a complex throughout its purification and gel filtration but its components are readily separated by electrophoresis in denaturing buffer. The 27 000 molecular weight band was selectively autophosphorylated when the enzyme was incubated in the presence of [-32P]ATP.When casein was used as substrate, physiological concentrations of naturally occurring polyamines such as spermine and spermidine markedly stimulated enzyme activity. However with phosvitin as substrate polyamines were strong inhibitors of the enzyme activity. This contrasting effect on intestinal kinase activity was also apparent using cytoplasmic proteins as endogenous phosphate acceptors. A characterization of this differential effect is presented and some possible physiological implications are discussed. 相似文献
42.
Synthesis and phosphorylation of cytoskeleton components in foetal,regenerating and adult normal rat hepatocytes during culture 总被引:1,自引:0,他引:1
Georges Baffet Annie Ruelland Bruno Clement Elisabeth Le Rumeur Siegmund Fischer 《Molecular and cellular biochemistry》1985,68(2):97-105
Summary Detergent insoluble material (DIM) was prepared by gentle treatment with detergent from foetal, regenerating and adult normal rat hepatocytes cultured for various times. It retained to some degree the morphology of the cells. After incubation of intact cells with 35S-methionine, most of the labelled DIM proteins were found to be components of the cytoskeleton. They included several cytokeratins, vimentin and actin. The synthesis rate varied with the age of animals and culture conditions. The high synthetic rate of vimentin in foetal and regenerating hepatocytes could be associated with cell proliferation. No correlation was found between cytokeratin synthesis and hepatocyte growth. Most of the cytoskeleton proteins could be phosphorylated in intact cells and in DIM from cultured hepatocytes. However the degree of phosphorylation of these proteins was not related to their synthetic rate. The decreased phosphorylation level in cultured adult rat hepatocytes could be related to the rapid loss of specific functions. 相似文献
43.
Gabriele Mezzetti Mariastella Moruzzi Giorgio Piccinini Maria G. Monti Bruno Barbiroli 《Molecular and cellular biochemistry》1986,70(2):141-149
Summary Chick duodenal mucosa contains an endogenous factor which is capable to inhibit selectively a homologous polyamine-sensitive protein kinase. The inhibitor was partially purified and characterized, and it was found to contain typical mucopolysaccharidic components.Glycosidases digestion studies, selective degradation analysis and spectrophotometric titrations with metachromatic dyes indicated that the inhibitor preparation contained two major moieties identified as heparin-like and heparan sulfate-like structures. In chick intestine the inhibitor was specific for polyamine-sensitive protein kinase since selectively interacted with it and was inert towards other cAMP-independent and cAMP-dependent protein kinases. The inhibitory effect of the endogenous factor was counteracted by naturally occurring polyamines such as spermine. The order of potency of various polyamines was: spermine > thermine spermidine diamines. The release of inhibition by addition of physiological concentrations of spermine was also apparent when using cytosolic proteins as endogenous phosphate acceptors. These results suggest that a possible role of polyamine in the regulation of polyamine-sensitive protein kinase in the intestine is to protect the enzyme from the inhibitory action of endogenous heparinoids. 相似文献
44.
Bruno Bjärnborg 《Hydrobiologia》1983,101(1-2):19-26
The seasonal variation in water chemistry was followed during 1980 and 1981 in four mountain brooks in the southern part of Swedish Lapland. In the area investigated the soil is calcareous and the brook water is very well buffered during the major part of the year, with alkalinity varying between 0.4 and 1.0 milliequivalents per liter and with pH values about 7.5. These years the snow had a pH of approximately 5.2, which was considerably higher than has been reported from adjacent areas in the lower, coniferous region. During snowmelt the water discharge increased drastically, and although the net transport of bicarbonate increased, alkalinity showed low values due to dilution with meltwater. pH decreased, but not further than to 6.3–6.5, far from values reported in 1979 (pH less than 5), apparently due to the comparatively clean snow. A slight deficit in alkalinity, as compared to the nonmarine calcium and magnesium content, points to an acidification impact on the area. During maximum runoff some chemical variables, like aluminium, iron, nitrogen and phosphorus, behaved reversely to what might be expected during dilution and reached maxima in concentrations. It is concluded that the extreme runoff characteristics of high mountain areas make brook water more sensitive to acid precipitation than might be expected when regarding only the calcareous properties of soil and bedrock. 相似文献
45.
Structural Genes for a Newly Recognized Acetolactate Synthase in Escherichia coli K-12 总被引:23,自引:15,他引:8
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Maurilio De Felice John Guardiola Bruno Esposito Maurizio Iaccarino 《Journal of bacteriology》1974,120(3):1068-1077
Evidence is reported that shows the presence in Escherichia coli K-12 of a newly found acetolactate synthase. This enzyme is the product of two genes, ilvH and ilvI, both located very close to leu. Amber mutations have been found in both genes and therefore their products are polypeptides. Mutations in the ilvH gene cause the appearance of an acetolactate synthase activity which is relatively resistant to valine inhibition and can be separated by adsorption on hydroxylapatite from another activity present in the extract and more sensitive to valine inhibition than the former. A mutant altered in the ilvI gene was isolated among the revertants sensitive to valine inhibition of an ilvH mutant. Such a mutant lacks the resistant acetolactate synthase. A temperature-sensitive revertant of the ilvI mutant contained a temperature-sensitive acetolactate synthase. Thus ilvI is the structural gene for a specific acetolactate synthase. The activity of the ilvH gene product has been measured by adding an extract containing it to a purified ilvI acetolactate synthase, which, upon incubation, became more sensitive to valine inhibition. Conversely, a valine-sensitive acetolactate synthase (the product of the ilvH and the ilvI genes) became more resistant to valine inhibition upon incubation with an extract of a strain containing a missense ilvH gene product. 相似文献
46.
Bruno Agostini 《Histochemistry and cell biology》1974,40(3):215-220
Summary Electron microscopy of negatively stained plasma shows that intravenous administration of heparin to healthy humans results in a fast degradation of very low density lipoproteins (VLDL), which are very rich in triglycerides (50% w/w). VLDL are also rapidly digested by post-heparin lipolytic activity when serum from non-heparinized subjects is incubated at 27°C in vitro with post-heparin serum of healthy controls.Abbreviations VLDL
very low density lipoproteins
- LDL
low density lipoproteins
- HDL
high density lipoproteins
- LP-X
lipoprotein-X, abnormal lipoprotein characterizing obstructive jaundice 相似文献
47.
The kinetics of denaturation of DNA have been studied by relaxation techniques. Examination of the terminal relaxation times for a variety of DNA's under a variety of conditions has shown that DNA denaturation is principally a hydrodynamically limited process. Measurements within the helix–coil transition have demonstrated that the experimentally measured terminal relaxation times are a function of the following: (1) position in the helix–coil transition; (2) ionic strength of the solvent; (3) solvent viscosity; (4) DNA concentration; (5) molecular weight; (6) number and position of single-strand breaks. The dependence of the terminal relaxation time on the above mentioned factors can be attributed to hydrodynamic effects. Thus a hydrodynamic model for DNA unwinding is required. The model which best fits the data involves the assumption of a rotational frictional coefficient independent of molecular weight. This assumption is suggested by the fact that the relaxation time is proportional to the first power of the molecular weight. 相似文献
48.
Zusammenfassung Die indirekten und direkten Flugmuskelfasern der Wanderheuschrecke enthalten parallel und voneinander getrennt verlaufende Myofibrillen. In den interfibrillären Räumen liegen zahlreiche, säulenförmig aneinandergereihte Mitochondrion, ein dichtes endoplasmatisches Retikulum und Tracheolen. In Höhe der A-I-Band-Grenzen befindet sich ein charakteristisches retikuläres System, dessen Queranordnung als Diade zu bezeichnen ist. Die chromatinreichen Kerne liegen peripher dicht unter dem Sarkolemm.Sowohl bei den indirekten als auch bei den direkten Flugmuskelgruppen kommt es im Verlaufe des Dauerfluges zu Transformationen der Mitochondrien, die sich in einer erheblichen Schwellung und in einer Umwandlung der Cristae zu Tubuli äußern. Diese Umwandlung wird als intramitochondriale Kompensation aufgefaßt, um das Verhältnis von Oberfläche der inneren membranösen Phase zum Volumen der inneren wäßrigen Phase — d. h. die spezifische Oberfläche — möglichst konstant zu halten. Eine Vesikulation der Tubuli könnte der morphologische Ausdruck für eine Dekompensation sein, da dann der Zusammenhang der äußeren wäßrigen Phase verlorengeht.Die Mitochondrien verschiedener Muskeln reagieren nicht synchron auf die Dauerbelastung, sondern ausgesprochen belastungs- und funktionsspezifisch: Zunächst transformieren sich die Mitochondrien der indirekten, dann erst die der direkten Flugmuskelgruppen, und der anfängliche Schwirrflug geht während fortschreitenden Dauerfluges in einen Flatterflug über. Die unterschiedlichen Erschöpfungszeitpunkte der indirekten Flugmuskeln werden durch die jeweils spezifische Funktion und die ontogenetische Entwicklung erklärt. 相似文献
49.
Bruno M. Cormier 《CMAJ》1966,94(6):276-291
A historical and psychological study of genocide is presented in which special emphasis is given to modern manifestations of this phenomenon. The policy of eradication of the Jews by the Hitler regime is considered as an example of genocide in the twentieth century. The psychopathology of genocide and the complex relationships among aggressors, victims and witnesses are elaborated. Inferences are drawn from past and present patterns of genocide that may provide some leads to the future. It is now possible to envisage an all-consuming genocide unless nations can learn to live together, not necessarily in unity but in diversity. 相似文献
50.
Pharmacological Characterization of Somatostatin Receptors in the Rat Cerebellum During Development 总被引:5,自引:2,他引:3
Bruno J. Gonzalez Philippe Leroux Corinne Bodenant Pierre Braquet Hubert Vaudry 《Journal of neurochemistry》1990,55(3):729-737
Abstract: Somatostatin (SRIF) receptors (SRIF-Rs) are transiently expressed in a germinative lamina of the rat cerebellum, the external granule cell layer. The appearance of SRIF-Rs coincides with the expression of SRIF-like immunoreactivity in the cerebellum. However, the cellular location of SRIF-Rs does not overlap with the distribution of SRIF-like immunoreactivity, with the latter being restricted to ascending fibers arising from the brainstem, to perikarya within the white matter, and to some Purkinje cells. The characterization of SRIF-Rs in the immature (13–day-old) rat cerebellum was conducted by means of binding experiments in membraneenriched preparations and autoradiography, using two radioligands, [125I-Tyr0,D-Trp8]SRIF-14 ([125I-Tyr0,d -Trp8]S14) and I25I-SMS 204–090. The pharmacological profile of cerebellar SRIF-Rs was compared with that of adult cortical SRIF-Rs. Saturation studies performed in 13–day-old rat cerebellum showed that the A'D values for [125I-Tyr0,D-Trp8]S14 and 125I-SMS 204–090 binding were 0.35 ± 0.04 and 0.39 ± 0.01 nM, respectively. The corresponding Bmax values were 52.7 ± 4.8 and 49.9 ± 5.3 fmol/mg of protein, a result indicating that radioligands with high specific radioactivity (2,000 Ci/mmol) bind to a single class of high-affinity sites (SSI). Competition studies showed that different D-Trp-sub-stituted analogs displaced [125I-Tyr0,d -Trp8]S14 binding with Hill coefficients >1, a finding indicating the existence of different subtypes of binding sites. When [Tyr0,d -Trp8]S14 was used as a competitor, two sites were resolved by Scatchard analysis in both 13–day-old cerebellum and adult cerebral cortex. The higher-affinity sites correspond to the SSI subtype identified in saturation experiments, whereas the lower-affinity sites most likely correspond to the SS2 subtype. Ionic supplementation studies showed that divalent cations were required to obtain maximal specific binding on the SSI sites. In particular, Mn2+ was the most efficient cation for promoting binding of [125I-Tyr0,d -Trp8]S14. Addition of GTP to the incubation buffer induced a marked reduction of specific binding. The results obtained by membrane binding assays were similar to those obtained by quantitative autoradiography, a result indicating that the microenvironment of SRIF-Rs was preserved in both types of tissue preparations. Receptors expressed in the developing rat cerebellum exhibited the same KD and similar pharmacological profile as those observed in the adult rat cortex. These results show that SRIF-binding sites transiently expressed in the external granule cell layer of the cerebellum of young rats are indistinguishable from adult rat brain SRIF-Rs. The extremely high density of SRIF-Rs found in the external granule cell layer in 13–day-old rats suggests that SRIF may play a pivotal role in the proliferation and/or differentiation of these germinative cells. 相似文献