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1.
Aser Rothstein 《The Journal of general physiology》1974,64(5):608-621
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
2.
Wolf Dietrich Jeschke 《Planta》1972,106(1):73-90
Summary Barley roots grown on a nutrient solution containing 1 mM Na+ but no K+ are capable of a considerable Na+ transport via the symplasm of the root and the xylem vessels. K+ added to the medium surrounding the root cortex severely inhibits this transport after a lag period at a high rate constant (Fig. 3).It is likely that the fluxes of Na+ are changed drastically during this transition from low to high K+ status. Although originally limited to steady state fluxes, the extended method of efflux analysis for excised roots (Pitman, 1971) has been applied to the non-steady fluxes which occur upon the addition of K+ to the roots. It is shown that besides other changes the efflux of 22Na+ through the cortex of barley roots is stimulated instantaneously (Fig. 5) by the addition of K+ and presumably by an influx of K+ ions. From this a transient, K+-stimulated Na+ efflux at the plasmalemma of the cortical cells can be estimated. It amounts to 10.9 moles/g fw · h compared to the control efflux of 3.3 moles/g fw · h without K+.The stimulated efflux is attributed to a Na+ efflux pump at the plasmalemma and is thus related to the K-Na-selectivity of barley plants. The inhibition of the Na+ transport by K+ is probably a consequence of this increased efflux of Na+ from the symplasm through the root cortex. 相似文献
3.
Sodium Transport in Capillaries Isolated from Rat Brain 总被引:20,自引:12,他引:8
A. Lorris Betz 《Journal of neurochemistry》1983,41(4):1150-1157
Abstract: Brain capillary endothelial cells form a bloodbrain barrier (BBB) that appears to play a role in fluid and ion homeostasis in brain. One important transport system that may be involved in this regulatory function is the Na+,K+-ATPase that was previously demonstrated to be present in isolated brain capillaries. The goal of the present study was to identify additional Na+ transport systems in brain capillaries that might contribute to BBB function. Microvessels were isolated from rat brains and 22Na + uptake by and efflux from the cells were studied. Total 22Na + uptake was increased and the rate of 22Na + efflux was decreased by ouabain, confirming the presence of Na+,K+-ATPase in capillary cells. After inhibition of Na+,K+-ATPase activity, another saturable Na + transport mechanism became apparent. Capillary uptake of 22Na + was stimulated by an elevated concentration of Na +or H+ inside the cells and inhibited by extracellular Na+, H+, Li+, and NH4+. Amiloride inhibited 22Na + uptake with a Ki between 10?5 and 10?6M but there was no effect of 1 mM furosemide on 22Na+ uptake by the isolated microvessels. These results indicate the presence in brain capillaries of a transport system capable of mediating Na +/ Na + and Na +/H + exchange. As a similar transport system does not appear to be present on the luminal membrane of the brain capillary endothelial cell, it is proposed that Na +/H + exchange occurs primarily across the antiluminal membrane. 相似文献
4.
D.Grant Gall Daniel G. Butler Fern Tepperman J.Richard Hamilton 《生物化学与生物物理学报:生物膜》1974,339(3):291-302
A technique to measure Na+ efflux from isolated intestinal epithelial cells has permitted us to examine the mechanisms responsible for Na+ transport in absorptive cells without contamination by other cell types. We examined the effect of actively transported sugars on Na+ efflux from isolated rat jejunal epithelial cells to evaluate the mechanism by which actively transported non-electrolytes stimulate Na+ absorption. Glucose, galactose and 3-O-methylglucose, sugars known to be actively transported by the small intestine, stimulate total Na+ efflux from isolated epithelial cells. This stimulation results from an increase of active Na+ transport, since it is inhibited by ouabain. Glucose stimulation is significantly greater than that produced by galactose or 3-O-methylglucose, 2-Deoxyglucose, a sugar that is not actively transported, has no effect on total Na+ efflux from isolated cells. Phloridzin, which has no effect on Na+ efflux in a sugar-free medium, completely abolishes the effect of galactose. These findings (a) support the hypothesis that the increase in intestinal absorption of Na+ in the presence of actively transported non-electrolytes occurs by a transcellular route; and (b) are consistent with the ion-gradient model. The results are not compatible with the direct energy-coupling model. 相似文献
5.
R.M. Johnstone 《生物化学与生物物理学报:生物膜》1975,413(2):252-264
Gramicidin induces a marked Na+-dependent efflux of amino acids from Ehrlich cells. In absence of Na+, gramicidin does not alter the efflux. In presence of gramicidin, glycine efflux is inhibited by methionine and less so by leucine. Glycine efflux caused by HgCl2 is neither Na+ dependent nor inhibitable by amino acids. Neither efflux of inositol which is transported by an Na+-dependent route, nor efflux of several other solutes which are transported by Na+-independent routes, is affected by gramicidin. The antibiotic appears to permit a reversal in the direction of the operation of the Na+-dependent amino acid transport system. The increased efflux is partly, but not entirely, due to an increase in the cellular Na+ concentration and a reduction of the electrochemical potential difference for Na+. 相似文献
6.
Use of Na-Nuclear Magnetic Resonance To Follow Sodium Uptake and Efflux in NaCl-Adapted and Nonadapted Millet (Panicum miliaceum) Suspensions
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Cellular Na+ transport was followed in vivo by 23Na nuclear magnetic resonance (NMR) using anionic dysprosium-based shift reagents to resolve internal and external 23Na+ resonances. Proso millet (Panicum miliaceum) cell suspensions adapted for rapid growth on 130 mm NaCl had biphasic 23Na efflux kinetics when shifted to low Na+ medium, while nonadapted cells had little measurable Na+ efflux after preloading with 23NaCl. Uptake of 23Na was also observed using 23Na NMR. The resonance frequency of the external Na+-dysprosium (III) triphosphate, relative to that of the 23Na in the cells, was sensitive to pH, permitting the pH of the external medium to be followed during the course of in vivo experiments. 相似文献
7.
Janos K. Lanyi 《Journal of cellular biochemistry》1977,6(2):169-177
Cell envelope vesicles prepared from H. halobium contain bacteriorhodopsin and upon illumination protons are ejected. Coupled to the proton motive force is the efflux of Na+. Measurements of 22Na flux, exterior pH change, and membrane potential, ΔΨ (with the dye 3,3′-dipentyloxadicarbocyanine) indicate that the means of Na+ transport is sodium/proton exchange. The kinetics of the pH changes and other evidence suggests that the antiport is electrogenic (H+/Na+ > 1). The resulting large chemical gradient for Na+ (outside > inside), as well as the membrane potential, will drive the transport of 18 amino acids. The 19th, glutamate, is unique in that its accumulation is indifferent to ΔΨ: this amino acid is transported only when a chemical gradient for Na+ is present. Thus, when more and more NaCl is included in the vesicles glutamate transport proceeds with longer and longer lags. After illumination the gradient of H+ collapses within 1 min, while the large Na+ gradient and glutamate transporting activity persists for 10–15 min, indicating that proton motive force is not necessary for transport. A chemical gradient of Na+, arranged by suspending vesicles loaded with KCl in NaCl, drives glutamate transport in the dark without other sources of energy, with Vmax and Km comparable to light-induced transport. These and other lines of evidence suggest that the transport of glutamate is facilitated by symport with Na+, in an electrically neutral fashion, so that only the chemical component of the Na+ gradient is a driving force. The transport of all amino acids but glutamate is bidirectional. Actively driven efflux can be obtained with reversed Na+ gradients (inside > outside), and passive efflux is considerably enhanced by intravesicle Na+. These results suggest that the transport carriers are functionally symmetrical. On the other hand, noncompetitive inhibition of transport by cysteine (a specific inhibitor of several of the carriers) is only obtained from the vesicle exterior and only for influx: these results suggest that in some respects the carriers are asymmetrical. A protein fraction which binds glutamate has been found in cholate-solubilized H. halobium membranes, with an apparent molecular weight of 50,000. When this fraction (but not the others eluted from an Agarose column) is reconstituted with soybean lipids to yield lipoprotein vesicles, facilitated transport activity is regained. Neither binding nor reconstituted transport depend on the presence of Na+. The kinetics of the transport and of the competitive inhibition by glutamate analogs suggest that the protein fraction responsible is derived from the intact transport system. 相似文献
8.
Joseph T. Tupper 《生物化学与生物物理学报:生物膜》1975,394(4):586-596
In a previous study, evidence was presented for an external Na+-dependent, ouabain-insensitive component of Na+ efflux and an external K+-dependent component of K+ efflux in the Ehrlich ascites tumor cell. Evidence is now presented that these components are inhibited by the diuretic furosemide and that under conditions of normal extracellular Na+ and K+ they represent Na+-for-Na+ and K-+for-K+ exchange mechanisms. Using 86Rb to monitor K+ movements, furosemide is shown to inhibit an ouabain-insensitive component of Rb+ influx and a component of Rb+ efflux, both representing approx. 30% of the total fux. Inhibition of Rb+ efflux is greatly reduced by removal of extracellular K+. Furosemide does not alter steady-state levels of intracellular K+ and it does not prevent cells depleted of K+ by incubation in the cold from regaining K+ upon warming. Using 22Na to monitor Na+ movements, furosemide is shown to inhibit an ouabain-insensitive component of unidirectional Na+ efflux which represents approx. 22% of total Na+ efflux. Furosemide does not alter steady-state levels of intracellular Na+ and does not prevent removal of intracellular Na+ upon warming from cells loaded with Na+ by preincubation in the cold. The ability of furosemide to affect unidirectional Na+ and K+ fluxes but not net fluxes is consistent with the conclusion that these components of cation movement across the cell membrane represent one-for-one exchange mechanisms. Data are also presented which demonstrate that the uptake of α-aminoisobutyrate is not affected by furosemide. This indicates that these components of cation flux are not directly involved in the Na+-dependent amino acid transport system A. 相似文献
9.
T. I. Ivanova A. O. Sherstobitov G. P. Gusev 《Journal of Evolutionary Biochemistry and Physiology》2001,37(5):528-534
To study H+ transport, the lamprey red blood cells were acidified to pH 6.0 by a pretreatment with an ionophore, nigericin. Incubation of the acidified cells in NaCl-medium at pH 8.0 was accompanied by a rapid H+ efflux from the erythrocytes. There was a tenfold decrease of the H+ efflux rate on addition to NaCl-medium of dimethylamiloride or on replacing Na+ in the medium (KCl-medium, pH 8.0). A high rate of Na+ influx into the acidified erythrocytes occurred only in the presence of H+ gradient (pH medium 8.0), but not in its absence (pH medium 6.0). The Na+-dependent H+ efflux from the cells and H+-dependent Na+ influx into the cells were quantitatively similar (about 700 mmol/l cells/h). A rapid elevation of the intracellular Na+ concentration as measured by flame photometry was also observed during incubation of the acidified cells in NaCl-medium (pH 8.0). The H+-dependent Na+ influx and an increase of the Na+ content in the acidified cells were significantly inhibited by amiloride. The data obtained for the first time prove with certainty the presence of the Na+/H+ exchanger in erythrocytes of the river lamprey. 相似文献
10.
11.
Gary Rudnick 《Journal of bioenergetics and biomembranes》1998,30(2):173-185
Neurotransmitter transporters are essential components in the recycling of neurotransmitters released during neuronal activity. These transporters are the targets for important drugs affecting mood and behavior. They fall into at least four gene families, two encoding proteins in the plasma membrane and two in the synaptic vesicle membrane, although the known vesicular transporters have not all been cloned. Each of these transporters works by coupling the downhill movement of small ions such as Na+, Cl–, K+, and H+ to the uphill transport of neurotransmitter. Plasma membrane transporters move the transmitter into the cytoplasm by cotransport with Na+. Many transporters also couple Cl– cotransport to transmitter influx and these all belong to the NaCl-coupled family, although within the family the coupling stoichiometry can vary. Transporters for glutamate couple influx of this excitatory amino acid to Na+ and H+ influx and K+ efflux. Transporters in synaptic vesicles couple H+ efflux to neurotransmitter transport from the cytoplasm to the vesicle lumen. 相似文献
12.
Although both amiloride- and phloretin-sensitive Na+/Li+ exchange activities have been reported in mammalian red blood cells, it is still unclear whether or not the two are mediated
by the same pathway. Also, little is known about the relative contribution of these transport mechanisms to the entry of therapeutic
concentrations of Li+ (0.2–2 mm) into cells other than erythrocytes. Here, we describe characteristics of these transport systems in rat isolated hepatocytes
in suspension. Uptake of Li+ by hepatocytes, preloaded with Na+ and incubated in the presence of ouabain and bumetanide, comprised three components. (a) An amiloride-sensitive component, with apparent K
m
1.2 mm Li+, V
max
40 μmol · (kg dry wt · min)−1, showed increased activity at low intracellular pH. The relationship of this component to the concentration of intracellular
H+ was curvilinear suggesting a modifier role of [H+]
i
. This system persisted in Na+-depleted cells, although with apparent K
m
3.8 mm. (b) A phloretin-sensitive component, with K
m
1.2 mm, V
max
21 μmol · (kg · min)−1, was unaffected by pH but was inactive in Na+-depleted cells. Phloretin inhibited Li+ uptake and Na+ efflux in parallel. (c) A residual uptake increased linearly with the external Li+ concentration and represented an increasing proportion of the total uptake. The results strongly suggest that the amiloride-sensitive
and the phloretin-sensitive Li+ uptake in rat liver are mediated by two separate pathways which can be distinguished by their sensitivity to inhibitors and
intracellular [H+].
Received: 8 April 1999/Revised: 19 July 1999 相似文献
13.
Freeman W. Cope 《The Journal of general physiology》1967,50(5):1353-1375
The nuclear magnetic resonance (NMR) spectrum of Na+ is suitable for qualitative and quantitative analysis of Na+ in tissues. The width of the NMR spectrum is dependent upon the environment surrounding the individual Na+ ion. NMR spectra of fresh muscle compared with spectra of the same samples after ashing show that approximately 70% of total muscle Na+ gives no detectable NMR spectrum. This is probably due to complexation of Na+ with macromolecules, which causes the NMR spectrum to be broadened beyond detection. A similar effect has been observed when Na+ interacts with ion exchange resin. NMR also indicates that about 60% of Na+ of kidney and brain is complexed. Destruction of cell structure of muscle by homogenization little alters the per cent complexing of Na+. NMR studies show that Na+ is complexed by actomyosin, which may be the molecular site of complexation of some Na+ in muscle. The same studies indicate that the solubility of Na+ in the interstitial water of actomyosin gel is markedly reduced compared with its solubility in liquid water, which suggests that the water in the gel is organized into an icelike state by the nearby actomyosin molecules. If a major fraction of intracellular Na+ exists in a complexed state, then major revisions in most theoretical treatments of equilibria, diffusion, and transport of cellular Na+ become appropriate. 相似文献
14.
Summary The rate of Na+/Na+ exchange as measured with 24Na+ in Na+-rich cells of Chlorella pyrenoidosa is governed by a single rate constant and saturates with increasing external Na+ concentration. The K
mvalue for this process is 0.8 mM Na+ and the maximum rate of exchange in illuminated cells is about 5 pmoles cm-2 sec-1. These values contrast with a K
mof 0.18 mM K+ and maximum rate of about 17 pmoles K+·cm-2·sec-1 for net K+ influx. Although the Na+/Na+ exchange was only slightly sensitive to light it was inhibited by the uncouplers CCCP and DNP and by the energy transfer inhibitor DCCD. This inhibition of the rate of Na+/Na+ exchange was not accompanied by a loss of internal Na+. Both the effect of external K+ on 24Na+ influx into Na+-rich cells and the inhibition of net K+ uptake by the presence of external Na+ indicates that Na+/Na+ and K+/Na+ exchanges share the same carrier and that the external site of this carrier has a three to four times higher affinity for K+ over Na+. 相似文献
15.
Shmuel Muallem Timothy Beeker Stephen J. Pandol 《The Journal of membrane biology》1988,102(2):153-162
Summary The relative contributions of the Na+/Ca2+ exchange and the plasma membrane Ca2+ pump to active Ca2+ efflux from stimulated rat pancreatic acini were studied. Na+ gradients across the plasma membrane were manipulated by loading the cells with Na+ or suspending the cells in Na+-free media. The rates of Ca2+ efflux were estimated from measurements of [Ca2+]
i
using the Ca2+-sensitive fluorescent dye Fura 2 and45Ca efflux. During the first 3 min of cell stimulation, the pattern of Ca2+ efflux is described by a single exponential function under control, Na+-loaded, and Na+-depleted conditions. Manipulation of Na+ gradients had no effect on the hormone-induced increase in [Ca2+]
i
. The results indicate that Ca2+ efflux from stimulated pancreatic acinar cells is mediated by the plasma membrane Ca2+ pump. The effects of several cations, which were used to substitute for Na+, on cellular activity were also studied. Choline+ and tetramethylammonium+ (TMA+) released Ca2+ from intracellular stores of pancreatic acinar, gastric parietal and peptic cells. These cations also stimulated enzyme and acid secretion from the cells. All effects of these cations were blocked by atropine. Measurements of cholecystokinin-octapeptide (CCK-OP)-stimulated amylase release from pancreatic acini, suspended in Na+, TMA+, choline+, or N-methyl-d-glucamine+ (NMG+) media containing atropine, were used to evaluate the effect of the cations on cellular function. NMG+, choline+, and TMA+ inhibited amylase release by 55, 40 and 14%, respectively. NMG+ also increased the Ca2+ permeability of the plasma membrane. Thus, to study Na+ dependency of cellular function, TMA+ is the preferred cation to substitute for Na+. The stimulatory effect of TMA+ can be blocked by atropine. 相似文献
16.
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
17.
Effects of ouabain and low temperature on the sodium efflux pump in excised corn roots 总被引:3,自引:3,他引:0
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Ouabain (0.05 millimolar) and low temperature (4 C) both caused the tissue Na+ content of excised 5-day-old corn roots to increase, indicating that there is an inhibition of the Na+ efflux pump. Na+ efflux was measured utilizing three different methods. Each method gave similar results in terms of rate and ouabain sensitivity. With one of these methods, the compartmental efflux method, it was demonstrated that rates for Na+ efflux increase as the external Na+ concentration is increased; e.g. the efflux rates are 0.529, 1.78, and 3.64 microequivalents per gram fresh weight per hour for external NaCl concentrations of 1, 10, and 30 millimolar, respectively. The data indicate that the Na+ efflux pump is located in the plasmalemma of root cells. 相似文献
18.
Using dialysed squid axons we have been able to control internal and external ionic compositions under conditions in which most of the Na+ efflux goes through the Na+ pump. We found that (i) internal K+ had a strong inhibitory effect on Na+ efflux; this effect was antagonized by ATP, with low affinity, and by internal Na+, (ii) a reduction in ATP levels from 3 mM to 50 μM greatly increased the apparent affinity for external K+, but reduced its effectiveness compared with other monovalent cations, as an activator of Na+ efflux, and (iii) the relative effectiveness of different K+ congeners as external activator of the Na+ efflux, though affected by the ATP concentration, was not affected by the Na+/+ ratio inside the cells. These results are consistent with the idea that the same conformation of the (Na+ + K+)-ATPase can be reached by interaction with external K+ after phosphorylation and with internal K+ before rephosphorylation. They also stress a nonphosphorylating regulatory role of ATP. 相似文献
19.
Hexachlorophene-induced hemolysis, as studied by phase contrast microscopy, appeared to be a result of osmotic swelling. Both swelling and subsequent hemolysis were markedly delayed by addition of the non-penetrating solute sucrose to the incubation mixture. Binding studies indicated that hexachlorophene is associated primarily with the erythrocyte membrane, the remainder being found in the cytoplasm. Hexachlorophane induced a dose-dependent, first-order efflux of Na+ and K+ from red cells. The rates of hemolysis and K+ efflux induced by hexachlorophene were much greater than would be expected if this compound were acting simply as a metabolic inhibitor and/or an inhibitor of (Na+-K+-Mg2+)-ATPase. It is suggested that hexachlorophene induces the efflux of Na+ and K+ from red cells by directly altering the permeability of the cellular membrane. Further, hexachlorophene-induced hemolysis is probably a secondary event resulting from osmotic swelling subsequent to increased membrane permeability. 相似文献
20.
Passive efflux of42K or86Rb from differentiated mouse neuroblastoma cells in culture was stimulated up to 8-fold by 10?4 M veratridine. The increased efflux could be blockedby low concentrations of tetrodotoxin (Ki = 4×10?9 g/ml), and did not occur with other cell types lacking an excitable membrane. The temperature sensitivity of the activated component was much higher than that of the normal passive outflow. It is suggested that the veratridine-dependent, tetrodotoxin-sensitive efflux represents passage of ions through the excitable Na+ channel. Replacement of extracellular Na+ by Tris+ abolished the activation by veratridine. Titration of the Na+ requirement resulted in a hyperbolic relationship between external Na+ concentration and efflux rate, with an apparent Km of 66.7 mM for Na+. This phenomenon may reflect an interaction between extracellular ions and a regulatory site on the Na+ channel. 相似文献