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1.
《Comparative biochemistry and physiology. A, Comparative physiology》1991,98(3):639-647
- 1.1. Gains in the water content of the Fundulus heteroclitus follicle account for over 90% of the total wet weight increase, and consequentially most of the size increase, associated with meiotic maturation.
- 2.2. Increases in intracellular Na+ and K+ actually exceed the accompanying increases in oocyte water, resulting in net gains in the concentration of these solutes. Changes in oocyte osmolality during maturation are mostly closely paralleled by variations in the concentration of K+.
- 3.3. Concentrations of various free amino acids, including taurine, remain constant or decline during maturation.
- 4.4. Taken together, these results suggest that an influx of K+, followed by osmotically-obliged water, is a primary cause of water uptake during oocyte maturation in F. heteroclitus.
2.
Romanova L. G. Novoselov S. V. Egorov M. V. Kostina M. B. Shakhparonov M. I. 《Russian Journal of Bioorganic Chemistry》2002,28(4):316-321
A comparative localization of Na+,K+-ATPase and ouabain-sensitive H+,K+-ATPase in rat skin was performed using in situ RNA hybridization and immunohistochemistry. Na+,K+-ATPase was predominantly detected in the basal layer of the epithelium, whereas the ouabain-sensitive H+,K+-ATPase, in the granular and prickle cell layers. The genes of these ATPases are thus expressed in epithelial cells at different stages of their development. The hypothesis was advanced that the ouabain-sensitive H+,K+-ATPase is involved in maintaining the skin pH value. The probes specific to the mRNAs of the full-size -subunit of the ouabain-sensitive H+,K+-ATPase and its truncated form were used to establish a similar distribution of both mRNA variants in skin. 相似文献
3.
Summary Delta endotoxin, a 68 kilodalton protein isolated fromBacillus thuringiensis spp.Kurstaki, is a potent entomocidal agent that alters a K+ current across midgut tissue of many phytophagous insects. This toxin completely inhibited the vanadate-sensitive86Rb+ uptake and mimicked the vanadate-induced decrease in cytosolic pH in a cell line (CHE) originating fromManduca sexta embryonic tissue. The toxin also inhibited a K+-sensitive-ATPase in the plasma membranes isolated from these cells. Using the K+-sensitive-ATPase substratp-nitrophenyl phosphate, delta endotoxin was found to have aK
i
of 0.4 m. These data suggest that the toxin inhibits a K+-ATPase responsible for86Pb+ uptake in the CHE cells. The relationship between the toxin inhibition of K+-ATPase and toxin-altered K+ current is discussed. 相似文献
4.
Charles Levinson 《The Journal of membrane biology》1987,100(1):183-191
Summary The volume regulatory response of the Ehrlich ascites tumor was studied in KCl-depleted, Na+-enriched cells. Subsequent incubation in K+-containing NaCl medium results in the reaccumulation of K+, Cl–, water and the extrusion of Na+. The establishment of the physiological steady state is due primarily to the activity of 2 transport systems. One is the Na/K pump (K
M for K
0
+
=3.5mm;J
max=30.1 mEq/kg dry min), which in these experiments was coupled 1K+/1 Na+. The second is the Cl–-dependent (Na++K+) cotransport system (K
M
for K
0
+
=6.8mm;J
max=20.8 mEq/kg dry min) which mediates, in addition to net ion uptake in the ratio of 1K+1Na+2Cl–, the exchange of K
i
+
for K
0
+
. The net passive driving force on the cotransport system is initially inwardly directed but does not decrease to zero at the steady state. This raises the possibility of the involvement of an additional source of energy. Although cell volume increases concomitant with net ion uptake, this change does not appear to be a major factor regulating the activity of the cotransport system. 相似文献
5.
Jochen Duhm 《The Journal of membrane biology》1987,98(1):15-32
Summary The effect of extracellular and intracellular Na+ (Na
o
+
, Na
i
+
) on ouabain-resistant, furosemide-sensitive (FS) Rb+ transport was studied in human erythrocytes under varying experimental conditions. The results obtained are consistent with the view that a (1 Na++1 K++2 Cl–) cotransport system operates in two different modes: modei) promoting bidirectional 11 (Na+–K+) cotransport, and modeii) a Na
o
+
-independent 11 K
o
+
/K
i
+
exchange requiring Na
i
+
which, however, is not extruded. The activities of the two modes of operation vary strictly in parallel to each other among erythrocytes of different donors and in cell fractions of individual donors separated according to density. Rb+ uptake through Rb
o
+
/K
i
+
exchange contributes about 25% to total Rb+ uptake in 145mm NaCl media containing 5mm RbCl at normal Na
i
+
(pH 7.4). Na+–K+ cotransport into the cells occurs largely additive to K+/K+ exchange. Inward Na+–Rb+ cotransport exhibits a substrate inhibition at high Rb
o
+
. With increasing pH, the maximum rate of cotransport is accelerated at the expense of K+/K+ exchange (apparent pK close to pH 7.4). The apparentK
m
Rb
o
+
of Na+–K+ cotransport is low (2mm) and almost independent of pH, and high for K+/K+ exchange (10 to 15mm), the affinity increasing with pH. The two modes are discussed in terms of a partial reaction scheme of (1 Na++1 K++2 Cl–) cotransport with ordered binding and debinding, exhibiting a glide symmetry (first on outside = first off inside) as proposed by McManus for duck erythrocytes (McManus, T.J., 1987,Fed. Proc., in press). N-ethylmaleimide (NEM) chemically induces a Cl–-dependent K+ transport pathway that is independent of both Na
o
+
and Na
i
+
. This pathway differs in many properties from the basal, Na
o
+
-independent K+/K+ exchange active in untreated human erythrocytes at normal cell volume. Cell swelling accelerates a Na
o
+
-independent FS K+ transport pathway which most probably is not identical to basal K+/K+ exchange. K
o
+
o
+
o
+
o
2+
reduce furosemide-resistant Rb+ inward leakage relative to choline
o
+
. 相似文献
6.
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
7.
Extrusion of H+ by intact barley (Hordeum vulgare L.) roots was automatically titrated. Simultaneously, uptake of K+ into the roots, transport of K+ through the roots, and (as a residual term) accumulation of K+ within the root tissue were determined. When no monovalent cation was present in the medium the steady rate of H+ release was close to zero. Addition of K+ stimulated H+ extrusion within less than 1 min. The stimulation of H+ release was apparently limited only by the movement of K+ through the apoplast of the roots. The steady rate of H+ extrusion depended on the availability of external K+ and saturated at a K+ concentration of about 100 mol· dm-3. Half-maximum rates of net K+ uptake and H+ extrusion were reached at a K+ concentration of about 10 mol·dm-3. With (slowly absorbable) sulfate as the only anion present, the stoichoimetry between H+ release and net K+ uptake was one. In conclusion, the uptake of K+ across the plasmalemma of the cells of the root cortex is electrically coupled to H+ extrusion. 相似文献
8.
Stephen D. McCormick 《Cell and tissue research》1990,260(3):529-533
Summary Anthroylouabain, a fluorescent derivative of ouabain, was used to localize Na+,K+-ATPase in transport epithelia of two species of teleosts. Exposure of the opercular membrane of seawater-adapted tilapia (Oreochromis mossambicus) and the jaw skin of the long-jawed mudsucker (Gillichthys mirabilis) to a 2 M anthroylouabain solution resulted in the appearance of cells stained bright blue. These were deemed to be chloride cells by their large size, distinct morphology and co-localization of DASPEI fluorescence, a mitochondrial stain. Addition of ouabain (1 mM final concentration) greatly decreased anthroylouabain fluorescent staining of chloride cells of seawater-adapted fish. Exposure of opercular membranes from freshwater tilapia to 2 M anthroylouabain did not result in significant staining. Anthroylouabain is therefore a useful vital stain for localizing Na+,K+-ATPase in chloride cells of seawater-adapted teleosts, and may be useful for fluorescent labelling of ouabain-sensitive Na+,K+-ATPase in other tissues and species. 相似文献
9.
Glen S. Patten Wayne R. Leifert Sharon L. Burnard Richard J. Head Edward J. McMurchie 《Molecular and cellular biochemistry》1996,154(2):133-141
Proton-dependent, ethylisopropylamiloride (EIPA)-sensitive Na+ uptake (Na+/H+ antiporter) studies were performed to examine if saliva, and ionophores which alter cellular electrolyte balance, could influence the activity of the cheek cell Na+/H+ antiporter. Using the standard conditions of 1 mmol/1 Na+, and a 65:1 (inside:outside) proton gradient in the assay, the uniport ionophores valinomycin (K+) and gramicidin (Na+) increased EIPA-sensitive Na+ uptake by 177% (p < 0.01) and 227% (p < 0.01), respectively. The dual antiporter ionophore nigericin (K+-H+) increased EIPA-sensitive Na+ uptake by 654% (p < 0.01), with maximal Na+ uptake achieved by 1 min and at an ionophore concentration of 50 mol/l, with an EC
50 value 6.4 mol/l. Preincubation of cheek cells with saliva or the low molecular weight (MW) components of saliva (saliva activating factors, SAF) for 2 h at 37°C, also significantly stimulated EIPA-sensitive Na+ uptake. This stimulation could be mimicked by pre-incubation with 25 mmol/l KCl or K+-phosphate buffer. Pre-incubating cheek cells with SAF and the inclusion of 20 mol/1 nigericin in the assay, produced maximum EIPA-sensitive Na+ uptake. After pre-incubation with water, 25 mmol/1 K+-phosphate or SAF, with nigericin in all assays, the initial rate of proton-gradient dependent, EIPA-sensitive Na+ uptake was saturable with respect to external Na+ with Km values of 0.9, 1.7, and 1.8 mmol/l, and V
max values of 13.4, 25.8, and 31.1 nmol/mg protein/30 sec, respectively. With 20 mol/1 nigericin in the assay, Na+ uptake was inhibited by either increasing the [K+]o in the assay, with an ID
50 of 3 mmol/l. These results indicate that nigericin can facilitate K+
i exchange for H+
o and the attending re-acidification of the cheek cell amplifies IINa+ uptake via the Na+/H+ antiporter. The degree of stimulation of proton-dependent, EIPA-sensitive Na+ uptake is therefore dependent, in part, on the intracellular K+
i. 相似文献
10.
Sergei N. Orlov Therese J. Resink Jens Bernhardt Fritz R. Buhler 《The Journal of membrane biology》1992,129(2):199-210
Summary To identify ion transport systems involved in the maintenance of vascular smooth muscle cell volume the effects of incubation medium osmolality and ion transport inhibitors on the volume and 86Rb and 22Na transport in cultured smooth muscle cells from rat aorta (VSMC) have been studied. A decrease of medium osmolality from 605 to 180 mosm increased intracellular water volume from 0.6 to 1.3 l per 106 cells. Under isosmotic conditions, cell volume was decreased by ouabain (by 10%, P< 0.005) but was not influenced by bumetanide, furosemide, EIPA and quinidine. These latter compounds were also ineffective in cell volume regulation under hypotonic buffer conditions. Under hyperosmotic conditions, cell volume was decreased by bumetanide (by 7%, P<0.05) and by ethylisopropyl amiloride (by 13%, P< 0.005). Ouabain-sensitive 86Rb influx was decreased by 30–40% under hypoosmotic conditions. An increase in medium osmolality from 275 to 410 mosm resulted in an eightfold increase in bumetanide-inhibited 86Rb influx and 86Rb efflux. The (ouabain and bumetanide)-insensitive component of 86Rb influx was not dependent on the osmolality of the incubation medium. However (ouabain and bumetanide)-insensitive 86Rb efflux was increased by 1.5–2 fold in VSMC incubated in hypotonic medium. Ethylisopropyl amiloride-inhibited 22Na influx was increased by sixfold following osmotic-shrinkage of VSMC. The data show that both Na+/H+ exchange and Na+/K+/2Cl– cotransport may play a major role in the regulatory volume increase in VSMC. Basal and shrinkage-induced activities of Na+/K+/2Cl– cotransport in VSMC were similarly sensitive to inhibition by either staurosporin, forskolin, R24571 or 2-nitro4-carboxyphenyl N,N-diphenylcarbomate (NCDC). In contrast basal and shrinkage-induced Na+/K+/2Cl– cotransport were differentially inhibited by NaF (by 30 and 65%, respectively), suggesting an involvement of guanine nucleotide binding proteins in the volume-sensitive activity of this carrier. Neither staurosporin, forskolin, R24571 nor NCDC influenced shrinkage-induced Na+/H+ exchange activity. NaF increased Na+/H+ exchanger activity under both isosmotic and hyperosmotic conditions. These data demonstrate that different intracellular signalling mechanisms are involved in the volume-dependent activation of the Na+/K+/2Cl– cotransporter and the Na+/H+ exchanger.The authors gratefully acknowledge the financial support of the Swiss National Foundation, grant No. 3.817.087. Bernadette Weber is thanked for preparing the figures. 相似文献
11.
Xylem parenchyma cells are situated around the (apoplastic) xylem vessels and are involved in the control of the composition of the xylem sap by exporting and resorbing solutes. We investigated properties of the K+ inward rectifier in the plasma membrane of these cells by performing patch clamp experiments on protoplasts in the whole-cell configuration. Inward currents were sensitive to the K+ channel blocker TEA+ at a high concentration (20 mm). Barium, another classical K+ channel blocker, inhibited K+ currents with a K
i
of about 1.3 mm. In contrast to guard cells, the cytosolic Ca2+ level proved to be ineffective in regulating the K+ conductance at hyperpolarization. External Ca2+ blocked currents weakly in a voltage-dependent manner. From instantaneous current-voltage curves, we identified a binding site in the channel pore with an electrical distance of about 0.2 to 0.5. Lanthanum ions reduced the inward current in a voltage-dependent manner and simultaneously displaced the voltage at which half of the channels are in the open state to more positive values. This finding was interpreted as resulting from a sum of two molecular effects, an interaction with the mouth of the channel that causes a reduction of current, and a binding to the voltage sensor, leading to a shielding of surface charges and, subsequently, a modulation of channel gating.A comparison between the K+ inward rectifier in xylem parenchyma cells, guard cells and KAT1 from Arabidopsis leads to the conclusion that these rectifiers form subtypes within one class of ion channels. The ineffectiveness of Ca2+ to control K+ influx in xylem parenchyma cells is interpreted in physiological terms. 相似文献
12.
Catecholamine- and volume-dependent ion fluxes in carp (Cyprinus carpio) red blood cells 总被引:1,自引:1,他引:0
S. N. Orlov G. A. Skryabin 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1993,163(5):413-420
In carp erythrocytes, noradrenaline (10-6 mol·l-1) induces a 30- to 40-fold activation of Na+/H+ exchange (the ethylisopropylamiloride-inhibited component of the 22Na influx) and a fourfold stimulation of the Na+, K+ pump (ouabain-inhibited component of 86Rb influx). In both cases the effect of noradrenaline is blocked by propranolol but not phentolamine and is imitated by forskolin. An activator of protein kinase C (-phorbol 12-myristate, 13-acetate) increases Na+/H+ exchange by 10 times and decreases the Na+, K+ pump activity by 20–30 percent. In the presence of ethylisopropylamiloride the increment of the Na+, K+ pump activity induced by noradrenaline is reduced by 35–45 percent, indicating the existence of a Na+/H+ exchange-independent mechanism of the Na+, K+ pump regulation by -adrenergic catecholamines. Hypertonic shrinkage of carp erythrocytes results in a 40- to 80-fold activation of Na+/H+ exchange, whereas hypotonic swelling induces an increase in the rate of 86Rb+ efflux which is inhibited by furosemide by about 30–40 percent. The rate of pH0 recovery in response to acidification or alkalinization in rat erythrocytes is approximately 15 times as fast as in carp erythrocytes. Unlike in rat erythrocytes, valinomycin does not cause an alkalinization of incubation medium in carp erythrocytes indicating the absence of conductive pathway in the operation of anion transporter protein. A scheme is suggested which describes the interrelation of Na+/H+ exchange, Na+, K+ pump and a non-identified system providing for K+ efflux in cell swelling, regulation of cell volume and cytoplasmic pH in fish erythrocytes under conditions of deep hypoxia and high activity.Abbreviations cAMP
cyclic adenosine monophosphate
- CCCP
carbonylcyamide m-chlorophenylhydrazone
- DMSO
dimethylsulphoxide
- EIPA
ethylisopropylamiloride
- NA
noradrenaline
- PMA
-phorbol 12-myristate, 13-acetate
- RVD
regulatory volume decrease
- RVI
regulatory volume increase 相似文献
13.
Fusicoccin (FC) has long been known to promote K+ uptake in higher plant cells, including stomatal guard cells, yet the precise mechanism behind this enhancement remains uncertain. Membrane hyperpolarization, thought to arise from primary H+ pumping stimulated in FC, could help drive K+ uptake, but the extent to which FC stimulates influx and uptake frequently exceeds any reasonable estimates from Constant Field Theory based on changes in the free-running membrane potential (V
m) alone; furthermore, unidirectional flux analyses have shown that in the toxin K+ (86Rb+) exchange plummets to 10% of the control (G.M. Clint and E.A.C. MacRobbie 1984, J. Exp. Bot.35 180–192). Thus, the activities of specific pathways for K+ movement across the membrane could be modified in FC. We have explored a role for K+ channels in mediating these fluxes in guard cells ofVicia faba L. The correspondence between FC-induced changes in chemical (86Rb+) flux and in electrical current under voltage clamp was followed, using the K+ channel blocker tetraethylammonium chloride (TEA) to probe tracer and charge movement through K+-selective channels. Parallel flux and electrical measurements were carried out when cells showed little evidence of primary pump activity, thus simplifying analyses. Under these conditions, outward-directed K+ channel current contributed appreciably to charge balance maintainingV
m, and adding 10 mM TEA to block the current depolarized (positive-going)V
m; TEA also reduced86Rb+ efflux by 68–80%. Following treatments with 10 M FC, both K+ channel current and86Rb+ efflux decayed, irreversbly and without apparent lag, to 10%–15% of the controls and with equivalent half-times (approx. 4 min). Fusicoccin also enhanced86Rb+ influx by 13.9-fold, but the influx proved largely insensitive to TEA. Overall, FC promotednet cation uptake in 0.1 mM K+ (Rb+), despite membrane potentials which were 30–60 mVpositive of the K+ equilibrium potential. These results tentatively link (chemical) cation efflux to charge movement through the K+ channels. They offer evidence of an energy-coupled mechanism for K+ uptake in guard cells. Finally, the data reaffirm early suspicions that FC alters profoundly the K+ transport capacity of the cells, independent of any changes in membrane potential.Abbreviations and symbols
E
K
equilibrium potential for K+
- FC
fusicoccin
- Hepes
4-(2-hydroxyethyl)-1-piperazineeth-anesulfonic acid
-
G
m
membrane (slope) conductance atV
m
- I-V
current-voltage (relationship)
-
apparent rate constant for exchange
- K
i
+
, K
0
+
intracellular, extracellular K+ (concentration)
- TEA
tetraethylammonium chloride
-
V
m
free-running membrane potential (difference) 相似文献
14.
Water and ion balance in hydrating oocytes of the grey mullet, Mugil cephalus (L.), during hormone-induced final maturation 总被引:1,自引:0,他引:1
Sequential changes in water content and inorganic ion concentrations occurring in intra-ovarian oocytes of the grey mullet, Mugil cephalus L., during hormone-induced final maturation are described. Post-vitellogenic oocytes have a water content of 59.4%. During final maturation, oocyte water increases rapidly to a maximum of 84.8% at oviposition. Hydration is accompanied by a relatively small increase in dry matter due to the concurrent uptake of inorganic ions and an apparent uptake of organic matter. Net uptake of Ca2+ during hydration is negligible. Net uptake of Mg2 + and Na+ occurs in less than equimolar proportions. Net uptake of K+ and Cl? occurs in greater than equimolar proportions prior to ovulation, returning to equimolarity at oviposition. Hence, hydration results in a dilution of intracellular Ca2+, Mg2+ and Na+ to minimum concentrations at oviposition. In contrast, K+ and Cl? concentrations are slightly elevated prior to ovulation, returning to initial levels at oviposition. A 1:1 stoichiometric relationship in net uptake of K + andCl? is evident. Intracellular osmotic pressure in post-vitellogenic oocytes is 203.4 ± 18.0 mOsmolkg?1. Osmotic pressure rises to a peak of 421.3+ 17.3 mOsmol kg?1 prior to ovulation, then declines to 313.3 ± 15.0 mOsmol kg?1 at oviposition. This rise in osmotic pressure prior to ovulation cannot be accounted for solely on the basis of inorganic ion uptake and must be attributed to changes in intracellular concentrations of organic solutes as well. An apparent increase in non-ash dry matter during hydration suggests that a net uptake of organic solute is involved. A relatively small organic molecule with high osmotic activity (e.g., amino acids) is implicated. 相似文献
15.
The preincubation of Chlorella cells with glucose caused a tenfold increase of the maximal uptake rate of ammonium without change in the K
m (2 M). A similar stimulation of ammonium uptake was found when the cells were transferred to nitrogen-free growth medium. The time-course of uptake stimulation by glucose revealed a lag period of 10–20 min. The turnover of the ammonium transport system is characterized by a half-life time of 5–10 h, but in the presence of light 30% of uptake activity stayed even after 50 h. 6-Deoxyglucose was not able to increase the ammonium uptake rate. These data together were interpreted as evidence for induction of an ammonium transport system by a metabolite of glucose. Mechanistic studies of the ammonium transport system provided evidence for the electrogenic uptake of the ammonium ion. The charge compensation for NH
4
+
entry was achieved by immediate K+ efflux from the cells, and this was followed after 1 min by H+ extrusion. Ammonium accumulated in the cells; the rate of uptake was sensitive to p-trifluoromethoxy-carbonylcyanide-phenylhydrazon and insensitive to methionine-sulfoxime. Uptake studies with methylamine revealed that methylamine transport is obviously catalyzed by the ammonium transport system and, therefore, also increased in glucose-treated Chlorella cells.Abbreviation p.c.
packed cells 相似文献
16.
Plant roots accumulate K+ over a range of external concentrations. Root cells have evolved at least two parallel plasma-membrane K+ transporters which operate at millimolar and micromolar external [K+]: high-affinity K+ uptake is energised by symport with H+, while low-affinity uptake is assumed to occur via ion channels. To determine the role of ion channels in low-affinity K+ uptake, a characterisation of the principal K+-selective ion channels in the plasma membrane of Arabidopsis thaliana (L.) Heynh. cv. Columbia roots was undertaken. Two classes of K+-selective channels were frequently observed: one inward (IRC) and one outward (ORC) rectifying with unitary conductances of 5 pS, 20 pS (IRCs) and 15 pS (ORC), measured in symmetrical 10 mM KCl. The dominant IRC (5 pS) and ORC (15 pS) were highly cation-selective (PCl PK < 0.025) but less selective amongst monovalent cations (PNaPK0.17–0.3). Both the IRC and the ORC were blocked by Ba2+, Cs+ and tetra-ethyl-ammonium, whereas 4-aminopyridine and quinidine selectively inhibited the ORC. The ORC open probability was steeply voltage-dependent and ORC activation potentials were close to the potassium equilibrium potential (EK+), enabling ORCs to conduct mainly outward, but occasionally inward, K+ current. By contrast, gating of the 5-pS IRC was weakly voltageependent and IRC gating was invariably restricted to membrane potentials more negative than EK+, ensuring K+ transport was always inwardly directed. Studies on channel activity were conducted for a large number of root cells grown at two levels of external [K+], one where K+ uptake is likely to be principally through channels (6 mM K+) and one where it must be energised (100 M K+). Shifting growth conditions from high to low K+ did not affect single-channel properties such as conductance and selectivity, nor the manifestation of the ORC and 20-pS IRC, but led to enhanced activity of the 5-pS IRC. The enhanced activity of the 5-pS IRC was mirrored by a parallel increase in unidirectional 86Rb+ influx after low-K+ growth, clearly indicating a dominant role of this particular channel in K+ uptake at supra millimolar external [K+].Abbreviations EK+
potassium equilibrium potential
- Em
membrane potential
- HK
high [K+]
- IRC
inward rectifying channel
- LK
low [K+]
- ORC
outward rectifying channel
- TEA
tetra-ethyl-ammonium
Financial support was provided by the Biotechnology and Biological Sciences Research Council (Grant PG87/529) and by the European Union (Framework III, Biotechnology Programme). 相似文献
17.
Om Prakash Mishra Maria Delivoria-Papadopoulos Geraldine Cahillane L. Craig Wagerle 《Neurochemical research》1989,14(9):845-851
The effect of lipid peroxidation on the affinity of specific active sites of Na+, K+-ATPase for ATP (substrate), K+ and Na+ (activators), and strophanthidin (a specific inhibitor) was investigated. Brain cell membranes were peroxidized in vitro in the presence of 100M ascorbate and 25M FeCl2 at 37°C for time intervals from 0–20 min. The level of thiobarbituric acid reactive substances and the activity of Na+, K+-ATPase were determined. The enzyme activity decreased by 80% in the first min. from 42.0±3.8 to 8.8±0.9 mol Pi/mg protein/hr and remained unchanged thereafter. Lipid peroxidation products increased to a steady state level from 0.2±0.1 to 16.5 ±1.5 nmol malonaldehyde/mg protein by 3 min. In peroxidized membranes, the affinity for ATP and strophanthidin was increased (two and seven fold, respectively), whereas affinity for K+ and Na+ was decreased (to one tenth and one seventh of control values, respectively). Changes in the affinity of active sites will affect the phosphorylation and dephosphorylation mechanisms of Na+, K+-ATPase reaction. The increased affinity for ATP favors the phosphorylation of the enzyme at low ATP concentrations whereas, the decreased affinity for K+ will not favor the dephosphorylation of the enzyme-P complex resulting in unavailability of energy for transmembrane transport processes. The results demonstrate that lipid peroxidation alters Na+, K+-ATPase function by modification at specific active sites in a selective manner, rather than through a non-specific destructive process. 相似文献
18.
In suspension-cultured cells of Glycine max and Catharanthus roseus, marked callose synthesis can be induced by digitonin and chitosan. Leakage of a limited pool of electrolytes precedes callose formation, K+ representing the major cation lost. Poly-L-ornithine, as well as the ionophores A 23187 and ionomycin, also induces some callose synthesis but to a lesser extent. Digitonin increases the net uptake of Ca2+ from the external buffer with a time course parallel to callose synthesis but lagging behind the leakage of K+. Nifedipine partly blocks callose synthesis as well as the digitonin-induced increase in net Ca2+ uptake. Taken together, the data support the hypothesis that addition of the various substances might indirectly lead to membrane perturbation causing the common event of an increase in net Ca2+ uptake which results in callose deposition by a direct activition of the Ca2+-dependent and plasma-membane-located 1,3--glucan synthase. 相似文献
19.
Na+, K+-ATPase Isozyme Diversity; Comparative Biochemistry and Physiological Implications of Novel Functional Interactions 总被引:12,自引:0,他引:12
Mobasheri A Avila J Cózar-Castellano I Brownleader MD Trevan M Francis MJ Lamb JF Martín-Vasallo P 《Bioscience reports》2000,20(2):51-91
Na+, K+-ATPase is ubiquitously expressed in the plasma membrane ofall animal cells where it serves as the principal regulator of intracellularion homeostasis. Na+, K+-ATPase is responsible for generating andmaintaining transmembrane ionic gradients that are of vital importance forcellular function and subservient activities such as volume regulation, pHmaintenance, and generation of action potentials and secondary activetransport. The diversity of Na+, K+-ATPase subunit isoforms andtheir complex spatial and temporal patterns of cellular expression suggestthat Na+, K+-ATPase isozymes perform specialized physiologicalfunctions. Recent studies have shown that the subunit isoformspossess considerably different kinetic properties and modes of regulationand the subunit isoforms modulate the activity, expression and plasmamembrane targeting of Na+, K+-ATPase isozymes. This review focuseson recent developments in Na+, K+-ATPase research, and in particular reportsof expression of isoforms in various tissues and experiments aimed atelucidating the intrinsic structural features of isoforms important forNa+, K+-ATPase function. 相似文献
20.
The kinetics of K+ efflux across the membranes of i) wild-type Escherichia coli poisoned by the thiol reagent N-ethylmaleimide, ii) K+ retention mutants and iii) glutathione-deficient mutants, have revealed a common K+ leaky phenotype; it is characterized by a very high rate of K+ efflux. The results suggest that the products of kefB and kefC genes could encode two K+ channels, both gated by glutathione. The possible function of these K+ channels seems to be a K+ exit controlled by the redox state of the cell; indeed, it can be inferred from the effects of several oxidants and reductants that turning on and off of the K+ efflux mediated by the channels can be correlated with the redox state of glutathione. 相似文献