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1.
The diffusion coefficients D (cm2/s), of four monovalent cations K+, Na+, Rb+ and Cs+ and of Ca2+ have been measured in phosphatidylcholine/water lamellar phase as a function of phase hydration and temperature and in the presence of divalent cations. Diffusion rates vary strongly with phase hydration, between 10?7 and 10?6 cm2/s for monovalent and 10?8 and 10?7 for Ca2+. The activation energies obtained are relatively small (5–10 kcal/mol). As the phase water content increases, a series of diffusion sequences is obtained, corresponding to the sequences predicted by Eisenman's theory of alkali ion equilibrium selectivity.This diffusionnal selectivity, which depends exclusively upon non-equilibrium parameters (mobility) within the hydrophilic path is discussed in respect to current theories of pore selectivity.  相似文献   

2.
We present results of molecular dynamics simulations for diffusion of Na+ ion in water-filled carbon nanotubes (CNTs) at 25°C using the extended simple point charge water potential. Simulation results indicate the general trend that the diffusion coefficients of Na+ ion and water molecule in CNTs decrease with an increase in water density and are larger than those in the bulk solution. The average potential energies of ion–water and water–water, the radial distribution functions, the hydration numbers and the residence times of the hydrated water molecules are discussed. The classical solventberg picture describes Na+ ion in water adequately for systems with the small values of diffusion coefficients.  相似文献   

3.
Lettré cells maintain a plasma membrane potential near — 60mV, yet are scarcely depolarized by 80 mM Rb+ and are relatively impermeable to 86Rb+. They are depolarized by ouabain without a concomitant change in intracellular cation content. Addition of K+ to cells suspended in a K+ free medium, or of Na+ to cells in a Na+ free medium, hyperpolarizes the cells. They contain electroneutral transport mechanisms for Na+, K+ and H+ which can function as Na+:K+ and Na+:H+ exchanges. It is concluded that plasma membrane potential of Lettré cells, in steady-state for Na+ and K+, is produced by an electrogenic Na+ pump sustained by electroneutral exchanges, and restricted by anion leakage.  相似文献   

4.
This work presents a detailed kinetic study that shows the coupling between the E2→E1 transition and Rb+ deocclusion stimulated by Na+ in pig-kidney purified Na,K-ATPase. Using rapid mixing techniques, we measured in parallel experiments the decrease in concentration of occluded Rb+ and the increase in eosin fluorescence (the formation of E1) as a function of time. The E2→E1 transition and Rb+ deocclusion are described by the sum of two exponential functions with equal amplitudes, whose rate coefficients decreased with increasing [Rb+]. The rate coefficient values of the E2→E1 transition were very similar to those of Rb+-deocclusion, indicating that both processes are simultaneous. Our results suggest that, when ATP is absent, the mechanism of Na+-stimulated Rb+ deocclusion would require the release of at least one Rb+ ion through the extracellular access prior to the E2→E1 transition. Using vanadate to stabilize E2, we measured occluded Rb+ in equilibrium conditions. Results show that, while Mg2 + decreases the affinity for Rb+, addition of vanadate offsets this effect, increasing the affinity for Rb+. In transient experiments, we investigated the exchange of Rb+ between the E2-vanadate complex and the medium. Results show that, in the absence of ATP, vanadate prevents the E2→E1 transition caused by Na+ without significantly affecting the rate of Rb+ deocclusion. On the other hand, we found the first evidence of a very low rate of Rb+ occlusion in the enzyme–vanadate complex, suggesting that this complex would require a change to an open conformation in order to bind and occlude Rb+.  相似文献   

5.
6.
Abstract

We report results of molecular dynamics simulations of the limiting conductance of Na2+, Cl2?, Na°, and Cl° in supercritical water using the SPC/E model for water in conjuction with our previous study (Lee et al., Chem. Phys. Lett. 293, 289 (1998)). The behavior of the limiting conductances of Na2+ and Cl2? in the whole range of water density shows almost the same trend as those of Na+ and Cl?, but the deviation from the assumed linear dependence of limiting conductances of Na2+ and Cl2? on the water density is smaller than that of Na+ and Cl?. The ratio of the limiting conductance of the divalentions to that of the corresponding monovalentions over the whole range of water density is almost constant. In the cases of Na2+ and Cl2?, the dominating factor of the number of hydration water molecules around ions in the higher-density region and the dominating factor of the interaction strength between the ions and the hydration water molecules in the lower-density region are also found as was the cases for Na+ and Cl?. These factors, however, are not so strong as for the corresponding monovalent ions because the change in the energetics, structure, and dynamics are very small mainly due to the strong Coulomb interaction of the divalent ions with the hydration water molecules. The diffusion coefficient of Na° and Cl° monotonically increases with decreasing water density over the whole range of water density. The increase of the diffusion coefficient with decreasing water density is attributed only to the dramatic decrease of the hydration number of water in the first solvation shell around the uncharged species. Among the two important competing factors in the limiting conductance of Na+ and Cl?, the effect of the number of hydration water molecules around the uncharged species is the only existing factor over the whole range of water density since the interaction strength between the uncharged species and the hydration water molecules very small through the LJ interaction. This result has confirmed the dominating factor of the number of hydration water molecules around ions in the higher-density region in the explanation of the limiting conductance of Na+ and Cl? in supercritical water at 673 K.  相似文献   

7.
86Rb+ uptake by yeast was not only stimulated by Rb+ or K+ but also by Na+. The uptake of 22Na+ was enhanced by both Rb+ and K+, but not by Na+, which was inhibitory at all concentrations applied. Inhibition of 22Na+ uptake by inactive Na+ occurred in two phases: one phase refers to inhibition at low Na+ concentrations and the other to inhibition at high Na+ concentrations. Our results can be qualitatively described by a two-site transport mechanism, having two cation binding sites, which must be occupied with monovalent cations before transport can occur.  相似文献   

8.
Summary To study the physiological role of the bidirectionally operating, furosemide-sensitive Na+/K+ transport system of human erythrocytes, the effect of furosemide on red cell cation and hemoglobin content was determined in cells incubated for 24 hr with ouabain in 145mm NaCl media containing 0 to 10mm K+ or Rb+. In pure Na+ media, furosemide accelerated cell Na+ gain and retarded cellular K+ loss. External K+ (5mm) had an effect similar to furosemide and markedly reduced the action of the drug on cellular cation content. External Rb+ accelerated the Na+ gain like K+, but did not affect the K+ retention induced by furosemide. The data are interpreted to indicate that the furosemide-sensitive Na+/K+ transport system of human erythrocytes mediates an equimolar extrusion of Na+ and K+ in Na+ media (Na+/K+ cotransport), a 1:1 K+/K+ (K+/Rb+) and Na+/Na+ exchange progressively appearing upon increasing external K+ (Rb+) concentrations to 5mm. The effect of furosemide (or external K+/Rb+) on cation contents was associated with a prevention of the cell shrinkage seen in pure Na+ media, or with a cell swelling, indicating that the furosemide-sensitive Na+/K+ transport system is involved in the control of cell volume of human erythrocytes. The action of furosemide on cellular volume and cation content tended to disappear at 5mm external K+ or Rb+. Thein vivo red cell K+ content was negatively correlated to the rate of furosemide-sensitive K+ (Rb+) uptake, and a positive correlation was seen between mean cellular hemoglobin content and furosemide-sensitive transport activity. The transport system possibly functions as a K+ and waterextruding mechanism under physiological conditiosin vivo. The red cell Na+ content showed no correlation to the activity of the furosemide-sensitive transport system.  相似文献   

9.
Relationship of Cation Influxes and Effluxes in Yeast   总被引:2,自引:0,他引:2       下载免费PDF全文
The Na+ efflux from Na+-rich yeast cells into a cation-free medium is largely balanced by the excretion of organic anions. In the presence of Rb+, K+, or high levels of H+ (pH 3–4), the Na+ efflux is increased and the organic anion excretion is suppressed so that stoichiometric cation exchanges occur. H+ participates in the exchanges, moving into or out of the cells depending on the external pH and on the concentration of external Rb+(K+). The total cation efflux is dependent on the external Rb+ concentration in a "saturation" relationship, but the individual cations in the efflux stream are not. The discrimination factor in the efflux pathway between H+ and Na+ is very large (of the order of 10,000), and between Na+ and K+ considerable (of the order of 50). For the latter pair, the recycling of K+ from the cell wall space is an important factor in the discrimination. In addition, the Na+ efflux as a function of Na+ content follows a sigmoidal curve so that the discrimination factor is increased at high levels of cellular Na+. Although the influx and efflux pathways behave as a tightly coupled system, the mechanism of coupling is not entirely clear. A single system with different cation specificities and kinetic behaviors on the inside and outside faces of the membrane could account for the data.  相似文献   

10.
Summary The ion permeability of rabbit jejunal brush border membrane vesicles was studied by measuring unidirectional fluxes with radioactive tracers and bi-ionic diffusion potentials with the potential-sensitive fluorescent dye, diS–C3-(5). Tracer measurements provide estimates of the absolute magnitudes of permeability coefficients, while fluorescence measurements provide estimates of relative and absolute ion permeabilities. The magnitudes of the permeability coefficients for Na+, K+, Rb+, and Br were approximately 5 nanoliters/(mg protein × sec) or 10–5 cm/sec as determined by radioactive tracer measurements. The apparent selectivity sequence, relative to Na+, as determined by bi-ionic potential measurements was: F, isetheionate, gluconate, choline (<0.1)+(1.0)–(1.5)=NO 3 (1.5)–(2.3)+(2.4)+(2.5)+(2.6)+(3.9) 4 +(12)–(40). The origin of this selectivity sequence and its relationship to the ion permeability of the brush border membrane in the intact epithelium are discussed.  相似文献   

11.
Isolated muscle cells from adult rat heart have been used to study the relationship between myocardial glucose transport and the activity of the Na+/K+ pump. 86Rb+-uptake by cardiac cells was found to be linear up to 2 min with a steady-state reached by 40–60 min, and was used to monitor the activity of the Na+/K+ pump. Ouabain (10?3 mol/I) inhibited the steady-state uptake of 86Rb+ by more than 90%. Both, the ouabain-sensitive and ouabain-insensitive 86Rb+-uptake by cardiac cells were found to be unaffected by insulin treatment under conditions where a significant stimulation of 3-O-methylglucose transport occurred. 86Rb+-uptake was markedly reduced by the presence of calcium and/or magnesium, but remained unresponsive towards insulin treatment. Inhibition of the Na+/K+ pump activity by ouabain and a concomitant shift in the intracellular Na+:K+ ratio did not affect basal or insulin stimulated rates of 3-O-methylglucose transport in cardiac myocytes. The data argue against a functional relationship between the myocardial Na+/K+ pump and the glucose transport system.  相似文献   

12.
A Coulter-orifice pulse-height analyzer system was used to measure volume spectra of mammalian cells in suspension at different times after the addition of an equal volume of water. In appropriate hypotonic medium, cultured mammalian cells rapidly increase in volume and then shrink, more slowly, approaching their initial volumes within 20 to 30 minutes at 37.5°C. The shrinking phase was found to be reversibly inhibited by ouabain and inhibited in both K+-free and Na+-free solutions; neither choline+ nor Li+ could substitute for extracellular Na+ in supporting the shrinking phenomenon but Rb+ and Cs+ were fairly good substitutes for K+. Under conditions similar to those with which the shrinking phenomenon was observed with cultured cells, it was not found with either human or mouse red blood cells. Two methods were used to determine intracellular Na+ and K+ content in osmotically shocked cells and in unshocked controls. An isotope equilibration method was employed with L5178-Y mouse lymphoblasts and a chemical determination by flame photometry was used with Ehrlich ascites tumor cells. The K+ content was significantly reduced and the Na+ content was unchanged or somewhat increased in cells which had returned to their original volumes in hypotonic medium. The K+ content was even more reduced but the Na+ content was greatly increased in cells which were osmotically shocked in the presence of ouabain.  相似文献   

13.
Summary The loop diuretic bumetanide binds specifically to the Na/K/2Cl cotransporter of many cell types including duck erythrocytes. Membranes isolated from these erythrocytes retain the ability to bind bumetanide when cells are exposed to cotransport activity stimuli prior to membrane isolation. An extensive study of the effects of ions on specific [3H]bumetanide binding to such membranes is presented here and compared to the activity of these ions in supporting transport function in intact cells. Both Na+ and K+ enhanced bumetanide binding in a saturable manner consistent with a single-site interaction. The K m for each ion was dependent on the concentration of the other cation suggesting heterotropic cooperative interactions between the Na+ and K+ binding sites. Na+ and K+ were partially replaceable, with the selectivity of the Na+ site being Na+ > Li+ > NH 4 + ; N-methyl-d-glucamine+, choline+ and tetramethylammonium+ also supported a small amount of specific binding when substituted for Na+. The selectivity of the K+ site was K+ Rb+ > NH 4 + > Cs+; N-methyl-d-glucamine+, choline+ and tetramethylammonium+ were inactive at this site. The results of transport experiments revealed a slightly different pattern. Li+ could partially substitute for Na+ in supporting coteansport, but other monovalent cations were completely inactive. The order of potency at the K+ site was NH 4 + > K+ Rb+ > Cs+ other monovalent cations. The effect of Cl- on bumetanide binding was biphasic, being stimulatory at low [Cl-] but inhibitory at high [Cl-]. As this implies the existence of two Cl- binding sites (termed Cl H and Cl L for the high- and low- affinity sites, respectively) each phase was examined individually. Cl- binding to Cl H could be described by a rectangular hyperbola with a K m of 2.5 mm, while kinetic analysis of the inhibition of bumetanide binding at high [Cl-] revealed that it was of a noncompetitive type (K i = 112.9 mm). The selectivity of anion binding to the two sites was distinct. Cl H was highly selective with Cl- > SCN- > Br-; F-, NO 3 - , ClO 4 - , MeSO 4 - , gluconate- and SO 4 2- were inactive. The efficacy of anion inhibition of binding to Cl L was ClO 4 - > I- > SCN- > NO3 > Cl-; F-, MeSO 4 - , gluconate-, and SO 4 2- were inactive. Thus, Cl H is much more selective than Cl L and largely accounts for the specificity of the system with respect to anion transport. SO 4 - , NO 3 - , I-, SCN- and ClO 4 - did not support cotransport when bound to Cl L and the latter three anions were inhibitory. Mg2+ was found to stimulate binding at a narrowly defined peak around 1.5 mm, but was inhibitory at higher concentrations. Other divalent cations caused a similar inhibition of bumetanide binding but did not exert a stimulatory effect at 1.5 mm. Divalent cations have little effect on cotransport in intact cells at concentrations up to 20 mm, suggesting that their effects on diuretic binding reflect interactions at internally disposed sites. Bumetanide binding was optimal at a pH of 7.8–8.1 and declined sharply as the pH was lowered towards 6. The titration curve correlated well with the effect of pH on cotransport in intact cells; the inhibitory effect of low pH suggests that protonation of the cotransporter may inhibit its function.We thank Drs. Brad Pewitt, John Westley and Mrinalini Rao for discussion, Sara Leung and Artelia Watson for their excellent technical assistance, and Dr. R.J. Turner for his gift of [3H] bumetanide. This work was supported in part by Cystic Fibrosis Center grant #CF RO11 7-04.  相似文献   

14.
Rb+ transport in low-K+ cells of Neurospora crassa is biphasic, transport at millimolar Rb+ being added to a transport process which saturates in the micromolar range. Both processes exhibit Michaelis-Menten kinetics, but in the micromolar phase the kinetic parameters depend on the K+ content of the cell (the lower the K+ content the lower the Km and the higher the Vmax). Normal-K+ cells, suspended in a buffer with millimolar K+, do not present Rb+ transport in the micromolar range. Millimolar transport in these cells presents kinetics which depend on the K+ in buffer (the higher the K+ the higher the Km), although the K+ content of the cells is constant. Na+ inhibits competitively Rb+ transport in low-K+ and normal-K+ cells, but, even when the differences between the Rb+Km values are more than three orders of magnitude, the apparent dissociation constant for Na+ is the same, and millimolar, in both cases.  相似文献   

15.
The size of the cytoplasmic pools of Rb+ and Na+ in roots of barley (Hordeum vulgare L.) was estimated by two independent methods:
  • 1 . from the time curves of net Rb+ and Na+ uptake in low-salt roots;
  • 2 . from the kinetics of ion exchange in Rb+-and Na+-saturated roots.
The results indicate that in low-salt roots there is a lag-phase in vacuolar accumulation of Rb+ but probably not of Na+. At the same time part of the cytoplasmic Na+ pool seems to be excluded from rapid exchange. Both for Rb+ and Na+, flux reduction in saturated tissue appears to be more pronounced at the tonoplast than at the plasmalemma.  相似文献   

16.
Na+,K+-ATPase activity was monitored in MDCK kidney epithelial cell monolayers and in cell extracts as a function of cell density, cAMP elevation, and exposure to hexamethylene bisacetamide (HMBA) and dimethylsulfoxide (Me2SO). Ouabain-sensitive Na+,K+-ATPase and 86Rb+ uptake activities, and the number of [3H]-ouabain binding sites were maximal in subconfluent cultures and decreased accompanying the development of a confluent monolayer. A sodium pump density of 8 × 107 pumps/cell was estimated for subconfluent cultures, declining to 9 × 105 pumps/cell at confluence. Previous studies have shown that dibutyryl cyclic AMP (Bt2cAMP), 1-methyl-3-isobutylxanthine (IBMX), or the differentiation inducers HMBA and Me2SO, which also caused cAMP elevation, all stimulated dome formation, a visible manifestation of active transepithelial Na+ and water transport (Lever, 1979). In the present study, all of these inducers were found to elevate intracellular Na+ content, implicating this variable in control of induction of dome formation. Operationally, inducers could be divided into two classes. HMBA and Me2SO partially inhibited ouabain-sensitive 86Rb+ influx. Ouabain, at a concentration that caused partial sodium pump inhibition and increased intracellular Na+ content, was also effective as an inducer. The second class, exemplified by IBMX and Bt2cAMP caused a furosemide-sensitive increase in intracellular Na+ content. This class of inducers stimulated ouabain-sensitive 86Rb+ uptake, presumably by substrate effects due to increased Na+ levels. The Na+ or ATP activation of Na+,K+-ATPase activity assayed in cell-free extracts, the affinity of the transport system for Rb+ in intact cells and intracellular ATP levels were unchanged by inducer treatment. Elevation of intracellular Na+ concentration, either by cAMP-stimulated, furosemide-sensitive mechanisms or by partial inhibition of the sodium pump may stimulate the induction of dome formation in MDCK cells.  相似文献   

17.
The growth rate ofSaccharomyces cerevisiae was dependent on K+ content in culture medium in a certain range of K+ concentrations. Above the upper limit of the range, growth did not respond to K+ increase, and below the lower limit, yeast died. Rb+ and Na+ enhanced growth in the range of K+ dependence and decreased the K+ concentration below which cells died. Both Rb+ and Na+ became toxic above a certain Rb+/K+ and Na+/K+ cellular ratio.  相似文献   

18.
Summary It is shown that the ouabain-resistant (OR) furosemide-sensitive K+(Rb+) transport system performs a net efflux of K+ in growing mouse 3T3 cells. This conclusion is based on the finding that under the same assay conditions the furosemidesensitive K+(Rb+) efflux was found to be two- to threefold higher than the ouabain-resistant furosemide-sensitive K+(Rb+) influx. The oubain-resistant furosemide-sensitive influxes of both22Na and86Rb appear to be Cl dependent, and the data are consistent with coupled unidirectional furosemide-sensitive influxes of Na+, K+ and Cl with a ratio of 1 1 2. However, the net efflux of K+ performed by this transport system cannot be coupled to a ouabain-resistant net efflux of Na+ since the unidirectional ouabain-resistant efflux of Na+ was found to be negligible under physiological conditions. This latter conclusion was based on the fact that practically all the Na+ efflux appears to be ouabainsensitive and sufficient to balance the Na+ influx under such steady-state conditions. Therefore, it is suggested that the ouabain-resistant furosemide-sensitive transport system in growing cells performs a facilitated diffusion of K+ and Na+, driven by their respective concentration gradients: a net K+ efflux and a net Na+ influx.  相似文献   

19.
为了探讨油菜素内酯对植物耐盐性的调控,以甘蓝型油菜"南盐油1号"为试验材料,研究了外源24-表油菜素内酯(24-EBL)对100、200 mmol/L Na Cl胁迫下油菜幼苗干重(DW)、相对含水量(RWC)、渗透调节能力(OAA)、叶片气体交换参数、气孔限制值(Ls)等的调节效应,还测定了不同器官的Na+、K+、Cl-含量,并计算各器官的K+/Na+和SK,Na。结果表明:(1)在不同浓度的盐胁迫下,油菜幼苗DW显著下降,胁迫下外源喷施10-12、10-10、10-8、10-6mol/L 24-EBL作用下,油菜植株干重均不同程度的上升,且植株干重都在10-10mol/L 24-EBL(EBL2)处理下达到最大值,分别比100、200 mmol/L Na Cl胁迫下增加29%和20%。与对照相比,非盐胁迫下外源喷施10-12、10-10、10-8、10-6mol/L 24-EBL,油菜幼苗植株干重与对照相比均无显著变化。(2)不同Na Cl浓度胁迫下,油菜叶片的RWC显著下降,外施EBL2可显著提高油菜叶片的RWC和OAA。(3)不同浓度Na Cl胁迫下,油菜幼苗叶片净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)和蒸腾速率(Tr)均不同程度下降,而Ls显著上升,而外喷EBL2可不同程度的提高Pn、Gs、Ci、Tr,降低Ls。(4)与对照相比,Na Cl胁迫下油菜幼苗叶片、叶柄和根的Na+和Cl-含量均显著上升,Na Cl浓度愈高,Na+和Cl-含量上升愈显著。而K+含量均下降,外源EBL2可显著降低幼苗各器官的Na+和Cl-含量,对幼苗叶片K+含量没有影响,但提高了叶柄和根中的K+含量。上述表明,合适浓度的24-EBL外喷可明显提高油菜的耐盐水平,且不同浓度Na Cl胁迫下,最适24-EBL浓度均为10-10mol/L。主要是因为外源喷施24-EBL能显著改善离子稳态和渗透调节能力,从而改善盐胁迫下油菜幼苗的光合作用、水分状况,提高其耐盐性。而24-EBL对盐处理下油菜植株气孔限制的显著改善是其促进其光合、水分利用的重要原因,也是其对100 mmol/L Na Cl处理的油菜生长调控效果优于200 mmol/L Na Cl处理的重要原因之一。结果还显示,在叶片中,24-EBL外施可通过排Na+和Cl-来维持植株离子稳态,而对K+影响不大;在根、茎中可通过排Na+、排Cl-、吸K+维持稳态。  相似文献   

20.
Abstract

The alkali-ion binding properties of two natural depsipeptide ion carriers, enniatin B (EnB) and valinomycin (VM), are examined and compared by the empirical force field method. While VM has been shown to bind preferentially K+, Rb+, and Cs+ over Na+ in most solvents, EnB is considerably less specific.

We find that EnB forms two kinds of complexes, internal and external. In internal complexes, the ion binds to all six carbonyl oxygens, while in external ones, only three oxygens, preferentially those of the D-hydroxy-isovaleryl residues, are bound. The size of the internal cavity is best suited for Na+, while K+ and Rb+ squeeze in asymmetrically by distorting the molecule, and Cs+ not at all. External binding is much less specific. Since internal complexes possess much higher strain energies than external ones, the latter may be at least as stable as the former, even in fairly non-polar solvents.

VM is calculated to bind only internally, and with much less strain energy than EnB. The size of its internal cavity is well suited for binding the ions K+, Rb+, and Cs+, but is too big for Na+. The difference between the binding energies of Na+ and K+ is much smaller than that between the corresponding hydration enthalpies, thus explaining the binding preference for the latter ion.  相似文献   

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