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1.
2.
Copper is an essential yet toxic metal ion. To satisfy cellular requirements, while, at the same time, minimizing toxicity,
complex systems of copper trafficking have evolved in all cell types. The best conserved and most widely distributed of these
involve Atx1-like chaperones and P1B-type ATPase transporters. Here, we discuss current understanding of how these chaperones bind Cu(I) and transfer it to the
Atx1-like N-terminal domains of their cognate transporter. 相似文献
3.
Rita Padányi Yuning Xiong Géza Antalffy Krisztina Lór Katalin Pászty Emanuel E. Strehler ágnes Enyedi 《The Journal of biological chemistry》2010,285(41):31704-31712
The membrane localization of the plasma membrane Ca2+-ATPase isoform 2 (PMCA2) in polarized cells is determined by alternative splicing; the PMCA2w/b splice variant shows apical localization, whereas the PMCA2z/b and PMCA2x/b variants are mostly basolateral. We previously reported that PMCA2b interacts with the PDZ protein Na+/H+ exchanger regulatory factor 2 (NHERF2), but the role of this interaction for the specific membrane localization of PMCA2 is not known. Here we show that co-expression of NHERF2 greatly enhanced the apical localization of GFP-tagged PMCA2w/b in polarized Madin-Darby canine kidney cells. GFP-PMCA2z/b was also redirected to the apical membrane by NHERF2, whereas GFP-PMCA2x/b remained exclusively basolateral. In the presence of NHERF2, GFP-PMCA2w/b co-localized with the actin-binding protein ezrin even after disruption of the actin cytoskeleton by cytochalasin D or latrunculin B. Surface biotinylation and fluorescence recovery after photobleaching experiments demonstrated that NHERF2-mediated anchorage to the actin cytoskeleton reduced internalization and lateral mobility of the pump. Our results show that the specific interaction with NHERF2 enhances the apical concentration of PMCA2w/b by anchoring the pump to the apical membrane cytoskeleton. The data also suggest that the x/b splice form of PMCA2 contains a dominant lateral targeting signal, whereas the targeting and localization of the z/b form are more flexible and not fully determined by intrinsic sequence features. 相似文献
4.
Makoto Tajima Nobuko Sekiguchi Masao Fujimaki 《Bioscience, biotechnology, and biochemistry》2013,77(2):319-320
High phosphate accumulating bacteria were isolated by autoradiography. One isoate, Arthrobacter globiformis PAB-6 accumulated phosphate intracellularly at 20% of dry cell mass in a simple synthetic medium. This amount was 3~7 times higher than type cultures examined. Almost no phosphate was released into the medium after cessation of growth. Fifty percent of total intracellular phosphate was fractionated as nucleic acids, while 20% each was recovered from cold PCA soluble fractions and polyphosphate fractions. The large content of nucleic acids in this bacterium appeared due to increased RNA content, specifically 4 S RNA fraction. 相似文献
5.
Ed Echeverria 《Physiologia plantarum》1991,81(1):51-54
The sucrose content of acid lime [ Citrus aurantifolia (Christm.) Swing.] juice tissue was measured at time 0 and at various times following incubation at 15.5, 26.6 and 37.7°C. The decline in sucrose content in fruit stored at 15.5°C paralleled the expected values for a sucrose solution at pH 2.1. At higher temperatures, the in vivo sucrose content decreased at significantly lower rates than the expected values. In fruit stored at 26.6 and 37.7°C, the vacuolar pH increased 0.11 and 0.23 units, respectively. When sucrose hydrolysis was recalculated at the increased vacuolar pH of juice cells stored at 26.6 and 37.7°C, the calculated values were similar to the measured values obtained in vivo. It is concluded that within the limits of the experimental conditions, the rates of sucrose acid hydrolysis are regulated by changes in the vacuolar H+ concentration. 相似文献
6.
The intensity and duration of the period of osmotic disturbance during introduction of brook charr into sea water were decreased by introducing the fish according to a gradient of salinity over a period of 6 days. Survival in summer increased from 25 to 90% with the use of a salinity gradient. However, kinetics and levels of activation of the gill Na+ , K+ -ATPase were not affected by the mode used for introducing brook charr into sea water. Neither was its level of activity modified by the use of a salted diet when the fish were in fresh water. The addition of 8 and 12% of salt to the diet prevented the plasma electrolyte surge of concentrations during the first days in sea water. In very cold water, survival rate was also drastically improved by giving an 8% salted diet during the 6 weeks preceding the introduction into sea water. These results show that both salty diets and exposure to brackish water during 6 days help brook charr face osmotic stress and improve their survival rate when introduced into full-strength sea water. The combined use of these preconditioning strategies might facilitate rearing this species in sea cages or silos. 相似文献
7.
A ribonuclease which was previously shown to be located in isolated vacuoles from suspension-cultured cells of tomato (Lycopersicon esculentum L.; Abel and Glund 1986, Physiol. Plant. 66, 79–86) has been purified to near homogeneity. Purification was up to 55000-fold with a yield of about 20%. The vacuolar origin of the protein was evidenced by comparing its electrophoretic mobility, isoelectric point, pH-optimum for activity and other properties with that of the RNA-degrading activity present in isolated vacuoles. The molecular weight of the native single polypeptide chain was estimated at 17500 and 20300 by gel filtration and sedimentation analysis, respectively. The enzyme hydrolyzed only single-stranded RNA with a mode of action that was endonucleolytic. The vacuolar ribonuclease had no requirement for divalent metal ions, and did not exhibit phosphomonoesterase (EC 3.1.3.1; EC 3.1.3.2) and phosphodiesterase (EC 3.1.15.1; EC 3.1.16.1) activity. The specificity of the enzyme has been studied by using homopolyribonucleotides as substrates. The end-products obtained were the respective nucleoside 2:3-cyclic monophosphates and, to minor extents, the corresponding nucleoside 3(2)-monophosphates. According to these observations, the vacuolar ribonuclease from tomato can be classified as ribonuclease I (EC 3.1.27.1).Abbreviations DEAE
diethylaminoethyl
- RNase
ribonuclease
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
8.
Angela F. Dulhunty Michael R. C. Banyard C. Jill Medveczky 《The Journal of membrane biology》1987,99(2):79-92
Summary Four monoclonal antibodies against the calcium ATPase in sarcoplasmic reticulum (SR) of rabbit fast-twitch skeletal muscle were characterized using SDS-PAGE, Western blots and immunofluorescence. The ultrastructural distribution of the antigens was determined using post-embedding immunolabeling. The antibodies recognized the calcium ATPase in the SR but not in transverse (T-) tubule or plasma membranes. The antibody, D12, had the same binding affinity for the calcium ATPase from fast-twitch (rabbit sternomastoid) and slow-twitch (rabbit soleus) fibers and the affinity fell by 30% after fixation for electron microscopy in both types of muscle fiber. Ultrastructural studies revealed that the density of D12 antibody binding to the terminal cisternae membrane of extensor digitorum longus (edl) and sternomastoid fibers was on average seven times greater than in the slow-twitch soleus and semimembranosus fibers. Since the affinity of the ATPase for the antibody was the same in SR from fast- and slow-twitch muscles, the concentration of calcium ATPase in the terminal cisternae membrane of fast-twitch fibers was seven times greater than in slow-twitch fibers. This conclusion was supported by the fact that the concentration of calcium ATPase in light SR membranes was six times greater in SR from fast-twitch fibers than in SR from slow-twitch fibers. The results provide strong evidence that the different calcium accumulation rates in mammalian fast- and slow-twitch muscles are due to different concentrations of calcium ATPase molecules in the SR membrane. 相似文献
9.
This study was undertaken in order to demonstrate the extent to which the activity of the plasmalemma H+-ATPase compensates for the charge and acidity flow caused by the sugar-proton symport in cells of chlorella vulgaris Beij.. Detailed analysis of H+ and K+ fluxes from and into the medium together with measurements of respiration, cytoplasmic pH, and cellular ATP-levels indicate three consecutive phases after the onset of H+ symport. Phase 1 occurred immediately after addition of sugar, with an uptake of H+ by the hexoseproton symport and charge compensation by K+ loss from the cells and, to a smaller degree, by loss of another ion, probably a divalent cation. This phase coincided with strong membrane depolarization. Phase 2 started approximately 5 s after addition of sugar, when the acceleration of the H+-ATPase caused a slow-down of the K+ efflux, a decrease in the cellular ATP level and an increase in respiration. The increased respiration was most probably responsible for a pronounced net acidification of the medium. This phase was inhibited in deuterium oxide. In phase 3, finally, a slow rate of net H+ uptake and K+ loss was established for several further minutes, together with a slight depolarization of the membrane. There was hardly any pH change in the cytoplasm, because the cytoplasmic buffering capacity was high enough to stabilize the pH for several minutes despite the net H+ fluxes. The quantitative participation of the several phases of H+ and K+ flow depended on the pH of the medium, the ambient Ca2+ concentration, and the metabolic fate of the transported sugar. The results indicate that the activity of the H+-ATPase never fully compensated for H+ uptake by the sugar-symport system, because at least 10% of symport-caused charge inflow was compensated for by K+ efflux. The restoration of pH in the cytoplasm and in the medium was probably achieved by metabolic reactions connected to increased glycolysis and respiration.Abbreviations DMO
dimethyloxazolidinedione
- EDTA
ethylcnediaminetetraacetic acid
- p.c.
packed cell volume 相似文献
10.
F Mercier H Reggio G Devilliers D Bataille P Mangeat 《Biology of the cell / under the auspices of the European Cell Biology Organization》1989,65(1):7-20
A monoclonal antibody (mab 146.14) marker of the movement of acid-secreting membranes in rat gastric parital cells has been produced and characterized. Mab 146.14 recognized a 95-kD major component of a purified membrane fraction of rat gastric mucosa, the protein composition of which was similar to that of well characterized porcine H+ -K+ ATPase-enriched membranes, and that presented the characteristic shift of density depending on whether it was purified from resting or stimulated tissues. Further biochemical analysis characterized the antigen as a membranous protein that might be in its native form, part of a higher multimolecular complex. Immunocytochemical localization of the antigen demonstrated that only membranes related to acid secretion in parietal cells expressed the 95-kD antigen. In resting conditions, the 95-kD antigen was diffusely distributed in the cell cytoplasm associated with inactive tubulovesicles. In stimulated cells, by contrast, all the antigen was recovered associated with secretory active microvilli formed by the apical insertion of the previously resting internal tubulovesicles. In conclusion, the 95-kD antigen, presumably a part of the rat gastric proton pump, is a marker of acid-secreting membranes in rat parietal cells. The translocation of antigen and membranes, observed by both light and electron microscopy supports the fusion model of membrane insertion from a cytoplasmic storage pool to the apical surface upon stimulation of acid secretion. 相似文献