SEARCH · PubMed Health

Results for “sodium pump”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 19 recordsLinked to original sources

PubMed1984

Effects of thyroid hormone on sodium pump sites, sodium content, and contractile responses to cardiac glycosides in cultured chick ventricular cells.

Sensitivity of cardiac muscle to digitalis glycosides depends on the thyroid state. The mechanism of this interaction was investigated at the cellular level using spontaneously beating monolayers of cultured chick embryo ventricular cells. Cells were grown for 48 h in serum-free medium containing concentrations of triiodothyronine (T3) from zero to 10(-7) M, and the total number of sodium pump sites, sodium content, and contractile amplitude in the presence and absence of various concentrations of ouabain were determined. T3 caused a concentration-dependent increase in the number of specific ouabain binding sites; the maximal increase to 160% of control was observed in response to 10(-8) M T3. T3 lowered steady-state cellular sodium content in a concentration-dependent manner, also. Ouabain (1 microM) exposure elevated cellular sodium content in all cells, but the increase was greatest in cells grown in T3-free medium and least in cells grown in 10(-8) M T3. The positive inotropic and toxic effects of ouabain in cells grown in 10(-8) M T3 were diminished at any given ouabain concentration, and thus, the dose-response curve was shifted to the right. These results indicate that T3 causes induction of additional sodium pump sites that are functional. The increased tolerance of hyperthyroid cells and reduced tolerance of hypothyroid cells to cardiac glycosides can be explained by these changes in the number of sodium pump sites and cellular sodium content, and consequently, calcium influx via sodium-calcium exchange.

Animals↗

PubMed2007

Sodium regulation, sodium pump function and sodium pump inhibitors in uncomplicated pregnancy and preeclampsia.

Preeclampsia is a disease characterized by hypertension and proteinuria but can manifest many abnormalities. Some of the best documented alterations involve changes in the handling of sodium ion both on the systemic and on the cellular level. There is broad agreement that the components of the renin-angiotensin-aldosterone pathway are markedly reduced in women with preeclampsia. However, other changes, especially those involving cell sodium are less consistent. A majority of studies support an increase in peripheral cell sodium concentration. This would suggest a defect in (Na,K)ATPase or sodium pump activity. Direct study of cellular sodium pump activity provides suggestive but not unequivocal support for this decreased sodium pump activity. Other evidence indicates increased circulating concentrations of a sodium pump inhibitor in most, but not all, studies of preeclampsia. Together, current research argues more strongly in favor of derangements of cell sodium handling perhaps mediated by circulating sodium pump inhibitors leading often to increased cell sodium. Such an increase of cell sodium in vascular tissue has previously been shown to enhance vascular sensitivity to vasoconstrictor agents or lead directly to increased vasoconstriction.

Biological Transport↗

PubMed1999

Platelet sodium pump and sodium potassium cotransport activity in nonpregnant, normotensive, and hypertensive pregnant women.

OBJECTIVE: To determine ouabain-sensitive sodium pump and bumetanide-sensitive sodium potassium cotransport activity in platelets from nonpregnant and normotensive pregnant women and from women with pregnancy-induced hypertension (PIH). METHODS: Blood was collected from 9 normotensive nonpregnant subjects, 24 normotensive pregnant subjects in both second and third trimesters, 9 subjects who developed proteinuric PIH, and 9 subjects who developed moderate nonproteinuric PIH. Platelet sodium pump activity was determined by the difference in the uptake of rubidium-86 in the presence and absence of ouabain; sodium potassium cotransport (SPC) activity is that component that is inhibitable by bumetanide. RESULTS: SPC activity was similar in normotensive subjects in the second [median (range) 78 mmol Rb/h/mg protein (18-140)] and third trimesters [85 (39-134)] but was significantly (p < 0.001) higher than in nonpregnant subjects [22 (4-107)]. In addition, SPC was significantly (p < 0.001) lower in subjects with nonproteinuric [42 (4-67)] or proteinuric PIH [59 (33-102)] compared to those who remained normotensive. Sodium pump activity was significantly higher (p < 0.05) in nonpregnant subjects [263 (188-430)] compared with the other groups of subjects. Total rubidium uptake was significantly higher (p < 0.05) in third-trimester normotensive subjects [471 (243-560)] compared with second-trimester subjects [405 (278-608)]. CONCLUSION: Our results suggest that the lower SPC activity in both nonproteinuric and proteinuric PIH may be an early sign of abnormality in the transport of sodium and potassium across the vascular smooth-muscle cell membrane, which is responsible for the maintenance of blood pressure.

Adult↗

PubMed1982

Sodium and potassium fluxes and membrane potential of human neutrophils: evidence for an electrogenic sodium pump.

Sodium and potassium ion contents and fluxes of isolated resting human peripheral polymorphonuclear leukocytes were measured. In cells kept at 37 degrees C, [Na]i was 25 mM and [K]i was 120 mM; both ions were completely exchangeable with extracellular isotopes. One-way Na and K fluxes, measured with 22Na and 42K, were all approximately 0.9 meq/liter cell water . min. Ouabain had no effect on Na influx or K efflux, but inhibited 95 +/- 7% of Na efflux and 63% of K influx. Cells kept at 0 degree C gained sodium in exchange for potassium ([Na]i nearly tripled in 3 h); upon rewarming, ouabain-sensitive K influx into such cells was strongly enhanced. External K stimulated Na efflux (Km approximately 1.5 mM in 140-mM Na medium). The PNa/PK permeability ratio, estimated from ouabain insensitive fluxes, was 0.10. Valinomycin (1 microM) approximately doubled PK. Membrane potential (Vm) was estimated using the potentiometric indicator diS-C3(5); calibration was based on the assumption of constant-field behavior. External K, but not Cl, affected Vm. Ouabain caused a depolarization whose magnitude dependent on [Na]i. Sodium-depleted cells became hyperpolarized when exposed to the neutral exchange carrier monensin; this hyperpolarization was abolished by ouabain. We conclude that the sodium pump of human peripheral neutrophils is electrogenic, and that the size of the pump-induced hyperpolarization is consistent with the membrane conductance (3.7-4.0 microseconds/cm2) computed from the individual K and Na conductances.

Biological Transport, Active↗

PubMed1994

A serial study of erythrocyte sodium pump kinetics and sodium content in the puerperium.

OBJECTIVE: Our purpose was to describe the alterations in erythrocyte sodium pump kinetics and sodium content occurring during the puerperium. STUDY DESIGN: Twelve healthy primigravid women were studied serially from late pregnancy until 20 weeks after delivery. Erythrocyte sodium pump rate constant, maximum velocity, and sodium affinity were calculated from the ouabain-sensitive sodium flux measured in whole blood and in erythrocytes in which sodium content had been altered with the ionophore nystatin. The Student t test was used to compare the regression coefficients of the values plotted against log time for specific periods. RESULTS: The sodium pump rate constant, maximum velocity, and sodium affinity were lower 20 weeks after delivery than in late pregnancy (0.339 +/- 0.018 vs 0.399 +/- 0.016/hr, 7.02 +/- 0.08 vs 9.98 +/- 0.078 mmol/kg/hr, 2.65 +/- 0.21 vs 3.16 +/- 0.20 mmol/kg). The decrease in the rate constant commenced after 4 days of the puerperium, whereas the decrease in maximum velocity and Michaelis-Menten constant did not commence until after 2 weeks. Erythrocyte sodium content was greater 20 weeks after delivery than in late pregnancy (4.71 +/- 0.20 vs 4.14 +/- 0.15 mmol/kg cells) and the increase was gradual over the time studied. CONCLUSIONS: After delivery the rate constant of the sodium pump measured in plasma and the erythrocyte sodium content changed before any significant alteration in the maximum velocity of the pump. The return of sodium pump function to the nonpregnant state continues beyond 6 weeks after delivery.

Erythrocytes↗

PubMed1981

Sulfatide role in the sodium pump.

Sodium efflux was studied in 22Na-loaded red blood cells in the presence of arylsulfatase, an enzyme that specifically hydrolyzes sulfatide. Sodium efflux was inhibited in proportion to the amount of arylsulfatase present. Maximum inhibition was almost as high as the efflux obtained in medium with K+ absent. At maximum inhibition 83.2% of the sulfatide content of the fragmented red blood cell membranes was hydrolyzed and ouabain-sensitive (Na+ + K+)-ATPase activity was inhibited by 100%. Sodium efflux, sulfatide content, and (Na+ + K+)-ATPase activity were unaffected with arylsulfatase in the presence of a high concentration of sulfatide. These results indicate that sulfatide plays a specific role in sodium and potassium ion transport. They also suggest that most sulfatide is localized externally in the red blood cell membrane.

Animals↗

PubMed1999

Sodium pump inhibition and regional expression of sodium pump alpha-isoforms in lens.

Both hypertension and cataract formation have been associated with reductions in sodium pump activity, possibly as a result of an endogenous inhibitor. The objective of the present study was to answer 4 closely related questions: (1) Is the lens sodium pump effectively inhibited by a labile, digitalis-like factor we have identified in the peritoneal dialysate from hypertensive patients in end-stage renal failure? (2) How does that inhibition compare to that induced by ouabain? (3) Does sodium pump isoform distribution determine the degree of lens sodium pump inhibition? (This question was precipitated by the unanticipated finding that the labile DLF was more effective in inhibiting lens sodium pump than was anticipated.) (4) Is sodium pump activity altered in lens in response to increased salt intake, a maneuver known to increase endogenous digitalis-like factor? We found that whereas ouabain produced equivalent or significantly less inhibition of lens Na(+), K(+)-ATPase from calf or rabbit, respectively, compared with brain, labile digitalis-like factor preferentially inhibited lens compared with brain. Analysis of whole-lens preparations from rabbit, calf, and normal human lens revealed substantial alpha2- and alpha3-isoforms of the sodium pump but little alpha1-isoform. Ouabain inhibition of whole-lens Na(+),K(+)-ATPase from rabbit and calf were comparable: for rabbit lens, K(i)=5.2x10(-7) mol/L; for calf lens, K(i)=1.0x10(-6) mol/L. Limited quantities of labile digitalis-like factor prohibited similar determinations; however, its concentration-activity profile paralleled that of ouabain. Na(+), K(+)-ATPase activity, measured in the 3 major anatomic regions of lens and normalized to nucleus, was greatest in epithelium (56. 9+/-17.9) compared with cortex (5.8+/-1.4) and nucleus (1.0+/-0.0; P=0.01). Immunohistochemistry of rabbit lens found abundant alpha2- and alpha3-isoforms in epithelium and limited alpha3 but undetectable alpha1 in cortex and nucleus. Finally, rats randomized to a high Na diet showed significantly reduced lens Na(+), K(+)-ATPase activity compared with those on a low Na diet, consistent with the effects of a sodium pump inhibitor. In conclusion, the present study suggests that digitalis-like factor may provide a link between hypertension and cataract formation.

Animals↗

PubMed1981

The magnesium dependence of sodium-pump-mediated sodium-potassium and sodium-sodium exchange in intact human red cells.

1. The magnesium content of human red blood cells was controlled by varying the magnesium concentration in the medium in the presence of the ionophore A23187. The new magnesium levels attained were very stable, which allowed the magnesium dependence of the sodium pump to be investigated.2. The effects of magnesium were shown to occur at the inner surface of the red cell membrane for the range of magnesium concentrations tested (10(-7) to 6 x 10(-3)m).3. At intracellular ionized magnesium concentrations below 0.8 mm the activation of ouabain-sensitive sodium-potassium exchange by internal ionized magnesium could be resolved into two or three components: (a) a small component, about 5% of the maximum flux, which is apparently independent of the ionized magnesium concentration below 2 mum, (b) a saturating component with a K((1/2)) of between 30 and 45 mum, and possibly (c) a component which increases linearly with ionized magnesium concentration and which only becomes apparent at concentrations above 0.1 mm.4. At intracellular ionized magnesium concentrations below 0.8 mm, activation of ouabain-sensitive sodium-sodium exchange by internal ionized magnesium could be resolved into two components: (a) a small component, about 6% of the maximal flux, which is apparently independent of the ionized magnesium concentration below 2 mum, and (b) a saturating component with a K((1/2)) of about 9 mum. At ionized magnesium concentrations between about 0.2 and 0.8 mm the rate of sodium-sodium exchange remained constant at the maximal level.5. The intracellular concentration of ATP decreased and the ADP concentration increased as the magnesium content of the cells was reduced from the normal level. A small increase in ATP and a small decrease in ADP was seen when the magnesium content was increased above the normal level. The variation in the ATP: ADP ratio from 2.5 at very low magnesium levels to about 6 at normal magnesium levels can account, at least in part, for the different K((1/2)) values of sodium-potassium and sodium-sodium exchange.6. When the concentration of ionized magnesium was increased above about 0.8 mm both sodium-potassium and sodium-sodium exchange were inhibited. Sodium-sodium exchange was more strongly inhibited than sodium-potassium exchange.7. The possible sites of action of magnesium in the sodium pump cycle are discussed.

Adenosine Diphosphate↗

PubMed2005

A third Na+-binding site in the sodium pump.

The sodium pump, or Na,K-ATPase, exports three intracellular sodium ions in exchange for two extracellular potassium ions. In the high resolution structure of the related calcium pump, two cation-binding sites have been identified. The two corresponding sites in the sodium pump are expected to be alternatively occupied by sodium and potassium. The position of a third sodium-specific site is still hypothetical. Here, we report the large effects of single residue substitutions on the voltage-dependent kinetics of the release of sodium to the extracellular side of the membrane. These mutations also alter the apparent affinity for intracellular sodium while one of them does not affect the intrinsic affinity for potassium. These results enable us to locate the third sodium-specific site of the sodium pump in a space between the fifth, sixth, and ninth transmembrane helices of the alpha-subunit and provide an experimental validation of the model proposed by Ogawa and Toyoshima [Ogawa, H. & Toyoshima, C. (2002) Proc. Natl. Acad. Sci. USA 99, 15977-15982].

Animals↗

PubMed2004

Recent insights into the structure and mechanism of the sodium pump.

The sodium pump (or Na-K-ATPase) is essential to the function of animal cells. Publication of the related calcium pump (SERCA) structure together with several recent results from a variety of approaches allow us to propose a mechanistic model to answer the question: "How does the sodium pump pump?"

Animals↗

PubMed2001

Expression of sodium pump isoforms and other sodium or calcium ion transporters in the heart of hypertensive patients.

The sodium pump (Na(+),K(+)-ATPase; EC 3.6.1.37) of animal cell membranes is the enzyme responsible for the maintenance of membrane potential, for the function of secondary active transporters, and for osmoregulation of the cell. Since inhibition of the enzyme by cardiac glycosides results in increased contractility of the heart muscle and increased blood pressure, we were interested in whether there is a correlation between hypertension and expression of the various isoforms of the sodium pump. In addition, we also examined the expression of the isoforms of the sarcoplasmic and plasma membrane Ca(2+)-ATPase, the Na(+)/Ca(2+)- and Na(+)/H(+)-exchangers, and Na(+) channel and Ca(2+) channel isoforms. Total mRNA was isolated from 50 mg tissue from the right atrium of hypertensive and normotensive patients who were undergoing cardiac surgery. After reverse transcription and subsequent amplification of ion transporter-specific cDNA fragments by polymerase chain reaction (PCR) in the presence of [alpha-(32)P]dCTP, quantification of the amplified fragments was carried out by the Phosphorimager technique. The data obtained show that the alphal subunit mRNA is expressed similarly in normotensive and hypertensive patients. The amount of alpha2 subunit mRNA, however, is increased 5-fold in hypertensive patients. In the same group, the amount of alpha3 isoform is also significantly increased, although not as dramatically as the alpha2 isoform. Besides the Na(+),K(+)-ATPase isoforms, a significant increase in the expression of mRNA for the Na(+)/Ca(2+)-exchanger and the plasma membrane Ca(2+)-ATPase isoforms was detected. It is possible that the observed changes in mRNA expression for these ion transporters reflect compensatory mechanisms to overcome a defective Na(+) and Ca(2+) metabolism in the tissues of hypertensive patients or reflect defects directly involved in the cause of hypertension. The expression of mRNA for all other transporters investigated was unaltered.

Adult↗

PubMed1980

Positive inotropic action of digoxigenin and sodium pump inhibition: effects of enhanced sodium influx.

A possibility that intracellular Na+ ions available to Na+,K+-adenosine triphosphatase influence the action of digoxigenin to cause sodium-pump inhibition and a positive inotropic effect was examined with isolated left atria of guinea-pig hearts. The positive inotropic action of digoxigenin developed more rapidly when atria were stimulated at 3 Hz than at 1.5 Hz. The rate of development of the positive inotropic action was dependent on the frequency of membrane depolarizations rather than on contractions. Monensin, a known Na+ ionophore, enhanced the rate of development of the positive inotropic action of digoxigenin. Sodium pump activity, as estimated from ouabain-sensitive 86Rb uptake, was inhibited by digoxigenin in a concentration-dependent manner in quiescent atria. The inhibition was enhanced by electrical stimulation which shifted the concentration-inhibition curves to the left. The sensitivity of the sodium pump for digoxigenin was also affected by membrane depolarizations, suggesting a role for intracellular Na+. These data indicate that similar to the cardiac glycosides, the interaction of the aglycone with Na+,K+-adenosine triphosphatase is essential for the development of the positive inotropic action of this agent.

Action Potentials↗

PubMed2004

Ouabain stimulates endothelin release and expression in human endothelial cells without inhibiting the sodium pump.

Ouabain, a sodium pump (Na+/ K+-ATPase) inhibitor, has been shown to act as a hormone and is possibly involved in the pathogenesis of hypertension. The mechanism by which ouabain may act was investigated using primary cultures of human umbilical artery endothelial cells (HUAECs), which are known to express and release the vasoconstrictive hormone endothelin (ET-1). Five minutes after application, low concentrations of ouabain induced Ca2+ oscillations and stimulated ET-1 release from endothelial cells into the medium. To investigate whether the observed effects were due to inhibition of the sodium pump, the effects of ouabain on the uptake of 86Rb+ by HUAECs were examined. Unexpectedly, ouabain concentrations below 10 nm stimulated 86Rb+ uptake by 15-20%, and in some experiments by 50%, results that are consistent with a stimulation of the pump. Within the concentration range 1-10 nm, ouabain induced a 2.5-fold stimulation (phosphorylation) of mitogen-activated protein kinase (MAP kinase). After incubation of HUAECs with ouabain for 12 h, the glycoside stimulated cell growth by 49 +/- 4%, as measured by cell number, with a maximum response at 5 nm. At similar concentrations, ouabain also increased ET-1 mRNA abundance by 19.5 +/- 3.1%. The results indicate that, by influencing ET-1 expression and release, ouabain may contribute to the regulation of vascular tone. The data also confirm that it is not a global inhibition of the sodium pump that is involved in the mechanism of action of this cardiac glycoside.

Animals↗

PubMed1985

Raised sodium pump activity and a circulating sodium transport inhibitor demonstrated on red blood cells of patients with untreated essential hypertension: correlation of pump activity with potassium permeability.

We have studied sodium potassium ATPase activity, the effect of endogenous plasma on sodium pump activity, potassium permeability and intracellular sodium and potassium concentrations in normotensive subjects without (n = 36) and with (n = 33) a positive family history of hypertension, and in patients with untreated essential hypertension (n = 52). Sodium pump activity was studied as ouabain sensitive uptake of rubidium 86 in washed red blood cells, incubated in an artificial medium closely resembling the anorganic constituents of plasma. Any influence of endogenous plasma on sodium pump activity was investigated by re-incubating the washed red blood cells in their own plasma and comparing ouabain sensitive rubidium uptake in the two media. To correct for any possible differences in external potassium concentration, a function for the relation between extracellular potassium concentration and absolute transport rates was derived experimentally. From this, actual transport rates in plasma were corrected by computer to an extracellular potassium concentration of 4.0 mmol/l. Sodium pump activity, concentration of circulating sodium transport inhibitor, potassium permeability and intracellular electrolytes were not statistically different in subjects with and without a positive family history of hypertension. Hypertensives had significantly raised sodium pump activity in artificial medium, but not when red cells were re-incubated in their own plasma. Thus, endogenous plasma inhibited the sodium pump by between 12% and 15%. Hypertensives also had a significantly raised potassium permeability. Potassium permeability and sodium pump activity were correlated significantly. Intracellular sodium concentrations were similar in normotensives and hypertensives, but the later showed a significantly lower intracellular potassium concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1985

Proton-activated rubidium transport catalyzed by the sodium pump.

Although the sodium pump normally exchanges three sodium for two potassium ions, experiments with inside-out red cell membrane vesicles show that the stoichiometry is reduced when the cytoplasmic sodium concentration is decreased to less than 1 mM. The present study was designed to gain insight into the question whether other monovalent cations, particularly protons, can act as sodium congeners in effecting pump-mediated potassium transport (ATP-dependent rubidium efflux from inside-out vesicles). The results show that at low cytoplasmic sodium concentration, an increase in proton concentration effects a further reduction in sodium:rubidium stoichiometry, to a value less than the minimal expected (1Na+:3Rb+). Furthermore, when vesicles containing 86RbCl are incubated in nominally sodium-free medium. ATP-dependent net rubidium efflux (normal influx) occurs when the pH is reduced from approximately 7.0 to 6.2 or less. This efflux is inhibited by strophanthidin and vanadate. These experiments support the notion that the sodium pump can operate as an ATP-dependent proton-activated rubidium (potassium) pump without obligatory countertransport of sodium ions.

Adenosine Triphosphate↗

PubMed1991

Sodium pump isozymes are differentially expressed in electrically dissimilar regions of colonic circular smooth muscle.

Molecular analyses of Na,K-ATPase abundance and alpha-subunit isoform distribution were performed to determine whether pump expression varies at different points through the thickness of the circular layer of colonic smooth muscle. The mRNA and polypeptides of Na,K-ATPase alpha 1 and beta subunits were twice as abundant in the submucosal region of the circular layer, which has previously been shown to generate large pump potentials. Sodium pump activity directly correlated with the relative abundance of the alpha 1 polypeptide. These data show that sodium pump expression varies in electrically dissimilar regions of the circular layer.

Animals↗

PubMed1985

Sodium pump inhibition, enhanced calcium influx via sodium-calcium exchange, and positive inotropic response in cultured heart cells.

The effects of sodium pump inhibition produced by exposure to the cardiac glycosides, ouabain or dihydroouabain, or by reduction in extracellular potassium to 1.0 mM, on contractile state and sodium-calcium exchange were studied in primary monolayer cultures of chick embryo ventricular cells. Ouabain, 10(-6)M, dihydroouabain, 5 X 10(-5)M, and extracellular potassium of 1.0 mM all induced similar and prominent positive inotropic effects. These effects were accompanied, in each case, by 40-50% inhibition of the rate of active uptake of 42K and by similar increases in steady state sodium content. Stimulation of the rate of 45Ca uptake on exposure to zero extracellular sodium occurred in response to extracellular potassium (1.0 mM) or to glycoside concentrations that induced a positive inotropic effect and sodium-potassium pump inhibition. Reactivation of the sodium pump after return from 1.0 to 4.0 mM extracellular potassium was rapid and was associated with membrane hyperpolarization and slowing of spontaneous beating rate. With pump reactivation under these circumstances, the time course of disappearance of stimulation of sodium-calcium exchange on exposure to zero extracellular sodium was similar to the time course of loss of the positive inotropic effect. Under physiological conditions (4.0 mM extracellular potassium), exposure to positively inotropic but nontoxic concentrations of ouabain or dihydroouabain caused a small but consistent increase in unidirectional calcium influx, but had no discernible effect on calcium efflux. Since similar inotropic effects were produced for comparable degrees of glycoside or low extracellular potassium-induced sodium pump inhibition and increases in cellular sodium content, sodium pump inhibition rather than a glycoside-specific change in calcium binding appears to underlie the inotropic response. These findings are further consistent with the view that the primary mechanism of the positive inotropic effects of digitalis and low extracellular potassium in this experimental preparation is sodium pump inhibition resulting in increased intracellular sodium. We suggest that increased calcium influx via sodium-calcium exchange is the principal mechanism whereby increased intracellular sodium results in enhanced calcium availability to the myofibrils, but an additional effect on calcium efflux is not excluded.

Animals↗

PubMed1993

Voltage-dependent inhibition of the sodium pump by external sodium: species differences and possible role of the N-terminus of the alpha-subunit.

Currents generated by the Na+/K+ ATPase were measured under voltage clamp in oocytes of Xenopus laevis. The dependence of pump current on external [Na+] was investigated for the endogenous Xenopus pump as well as for wild-type and mutated pumps of electroplax of Torpedo californica expressed in the oocytes. The mutants had alpha-subunits truncated before position Lys28 (alpha delta K28) or Thr29 (alpha delta T29) of the N-terminus. The currents generated by all variants of pump molecules in the presence of 5 mM K+ show voltage-dependent inhibition by external [Na+]. The apparent KI values increase with membrane depolarisation, and the potential dependence can be described by the movement of effective charges in the electrical potential gradient across the membrane. Taking into account Na(+)-K+ competition for external binding to the E2P form, apparent KI values and effective charges for the interaction of the Na+ ions with the E2P form can be estimated. For the Xenopus pump the effective charge amounts to 1.1 of an elementary charge and the KI value at 0 mV to 44 mM. For the wild-type Torpedo pump, the analysis yields values of 0.73 of an elementary charge and 133 mM, respectively. Truncation at the N-terminus removing a lysine-rich cluster of the alpha-subunit of the Torpedo pump leads to an increase of the effective charge and decrease of the KI value. For alpha delta K28, values of 0.83 of an elementary charge and 117 mM are obtained, respectively. If Lys28 is included in the truncation (alpha delta T29), the effective charge increases to 1.5 of an elementary charge and the apparent KI value is reduced to 107 mM. The KI values for pump inhibition by external Na+, calculated by taking into account Na(+)-K+ competition, are smaller than the K1/2 values determined in the presence of 5 mM [K+]. The difference is more pronounced for those pump variants that have higher Km values. The variations of the parameters describing inhibition by external [Na+] are qualitatively similar to those described for the stimulation of the pumps by external [K+] in the absence of extracellular [Na+]. The observations may be explained by an access channel within the membrane dielectric that has to be passed by the external Na+ and K+ ions to reach or leave their binding sites. The potential-dependent access and/or the interaction with the binding sites shows species differences and is affected by cytoplasmic lysine residues in the N-terminus.

Animals↗