Chromosomal Location of a Gene Involved in Potassium Ion Uptake in Escherichia coli B

A chromosomal lesion responsible for defective potassium ion uptake in Escherichia coli B has been mapped by use of standard interrupted-mating crosses. The mutation, kac-1, is in strain RD-2, which is deficient in K⁺ intake, exchanges cell K⁺ for extracellular isotope at a reduced rate, and has an abnormality of phosphorus metabolism associated with its potassium deficiency. This report places kac-1 at about 4 min clockwise from pro, close to gal. The locus of kac-1 is distinctly different from the potassium retention mutant in strain B-207, the only other potassium accumulation mutant mapped in E. coli so far. In this study, two other potassium accumulation mutations, kac-2 and kac-3, whose particular type of accumulation defects have not yet been determined, were mapped. These mutations are in the same region of the chromosome as kac-1.     

Potassium activation of Na+-dependent leucine transport in brush-border membrane vesicles from rat jejunum

Na⁺-dependent leucine uptake was greater in potassium loaded brush-border membrane vesicles compared with controls. This effect was not mediated by an electrical potential difference, since it was still present in voltage-clamped conditions. Inhibition experiments indicate the same Na⁺-dependent leucine transport activity in the presence or in the absence of potassium. The affinity of sodium for the cotransporter was identical at 10 or 100 mM potassium. Leucine kinetics at different potassium concentrations showed a maximum 2.4-fold increase in V_max, while K_m was unaffected. The secondary plots of the kinetic results were not linear. This kinetic behaviour suggests that K⁺ acts as a non-essential activator of Na⁺-dependent leucine cotransport. A charge compensation of sodium-leucine influx is most probably a component of the potassium effect in the presence of valinomycin.     

Modulation of active potassium transport by aldosterone

• 1.1. The role of aldosterone on active potassium transport across lizard colon under voltage-clamped conditions has been investigated.
• 2.2. Control colons exhibited no net potassium flux (J^k_net) despite of the existence of active opposite unidi ectional fluxes.
• 3.3. An important net secretory potassium flux was found in short-circuited aldosterone-stimulated colons.
• 4.4. Mucosal amiloride did not change (J^k_net) either in control or aldosterone-stimulated colons.
• 5.5. Luminal barium alters K ⁺ transport in a manner consistent with the presence of barium-sensitive conductances at the apical membrane of both control and aldosterone-treated colons.
• 6.6. The effects of ouabain and barium on control and aldosterone-induced potassium flows were consistent with a model involving basolateral uptake by an Na ⁺-K ⁺-ATPase and conductive exit across the apical membrane.
• 7.7. The stimulatory effect of aldosterone on potassium secretion is associated with parallel increases of both basolateral K ⁺ entry and the apical conductive pathway.

     

An electrogenic Na+/K+ pump in the choroid plexus

Intracellular electrical potential and potassium activity was measured by means of microelectrodes in the epithelial cells of choroid plexus from bullfrogs (Rana catesbeiana). Ouabain applied from the ventricular side caused an abrupt depolarisation of 10 mV but only a gradual loss of potassium from the cells. Readministration of potassium to the ventricular solution of plexuses which were previously depleted of potassium, caused a hyperpolarisation of about 4 mV. These two experiments are consistent with the notion of an electrogenic Na⁺/K⁺ pump situated at the ventricular membrane and which pumps potassium into the cell and sodium into the ventricle. The numerical values obtained suggest that 3 sodium ions are pumped for 2 potassium ions. The permeability coefficient for potassium exit from the cell is calculated to be 1.24 · 10^?5 cm^?1 · s^?1 expressed per cm² of flat epithelium.     

The Effect of Potassium on the Fixation of Molecular Nitrogen by Root Nodules of Vicia faba

The effect of potassium supply of Vicia faba on the fixation of molecular nitrogen by root nodules was studied by using ¹⁵N-labeled molecular nitrogen. Plants well supplied with potassium showed higher contents of ¹⁵N in the soluble amino fraction and in the protein fraction of various plant organs as compared with plants of a lower potassium status. This effect was evident particularly in the root nodules. Assimilation experiments, carried out with ¹⁴CO₂, revealed that the content of radioactivity in the sugars and amino acids of the root nodules was increased by the potassium supply of the host plants. In particular, the content of ¹⁴C amino acids in the root nodules was influenced beneficially by potassium, which means that potassium favored the provision of reduced nitrogen (NH₃). It is postulated that the better carbohydrate supply of nodules, by plants well supplied with potassium, results in a higher carbohydrate turnover in the nodules and thus the provision of ATP and reducing electrons required by the nitrogenase is enhanced.     

Potassium Ions and the Inhibitory Process in the Crayfish Stretch Receptor

The effect of the absence of potassium in the bathing solution on the synaptic inhibitory potentials of the crayfish stretch receptor has been studied. The inhibitory potentials were increased in size, i.e. became more hyperpolarizing, in the absence of potassium. Since the resting potential of the cell is increased in the absence of potassium, the alteration of the inhibitory potentials implies that the potassium conductance of the membrane is increased. While other ions, e.g. Cl^-, may also be involved, it seems that the membrane potential during inhibition is mainly dominated by K⁺.     

Regulation of Trk-dependent potassium transport by the calcineurin pathway involves the Hal5 kinase

The phosphatase calcineurin and the kinases Hal4/Hal5 regulate high-affinity potassium uptake in Saccharomyces cerevisiae through the Trk1 transporter. We demonstrate that calcineurin is necessary for high-affinity potassium uptake even in the absence of Na⁺ stress. HAL5 expression is induced in response to stress in a calcineurin-dependent manner through a newly identified functional CDRE (nt −195/−189). Lack of calcineurin decreases Hal5 protein levels, although with little effect on Trk1 amounts. However, the growth defect of cnb1 cells at K⁺-limiting conditions can be rescued in part by overexpression of HAL5, and this mutation further aggravates the potassium requirements of a hal4 strain. This suggests that the control exerted by calcineurin on Hal5 expression may be biologically relevant for Trk1 regulation.     

Potassium Ion Current in the Squid Giant Axon: Dynamic Characteristic

Measurements of the potassium current in the squid axon membrane have been made, after changes of the membrane potential to the sodium potential of Hodgkin and Huxley (HH), from near the resting potential, from depolarizations of various durations and amplitudes, and from hyperpolarizations of up to 150 mv. The potassium currents I given by I = I_∞ {1 - exp [- (t + t₀)/τ]}²⁵, where t₀ is determined by the initial conditions, represent the new data and approximate the HH functions in the regions for which they are adequate. A corresponding modification for the sodium current does not appear necessary. The results support the HH assumptions of the independence of the potassium and sodium currents, the dependence of the potassium current upon a single parameter determined by the membrane potential, and the expression of this parameter by a first order differential equation, and, although the results drastically modify the analytical expressions, they very considerably extend the range of apparent validity of these assumptions. The delay in the potassium current after severe hyperpolarization is used to estimate a potassium ion mobility in the membrane as 10^-5 of its value in aqueous solutions.     

Some electrophysiological and permeability properties of the mouse egg

Certain electrophysiological and ionic properties of the mouse egg (CF-1 and BDF 12–18 hr post ovulation) have been investigated. Membrane potential (?14 ± 0.4 mV, ± SE, inside negative), membrane resistance (2610 ± 38 ohm·cm²), and membrane capacitance (1.6 ± 0.03 μF cm^?2) have been determined by means of intracellular microelectrode recording techniques. Membrane potential and related parameters are stable for extended periods of time upon impalement and the magnitude of the cell membrane potential has been demonstrated to be sensitive to alteration in external sodium. The electrophysiological studies in conjunction with measurements of unidirectional potassium fluxes using isotope tracer-techniques have allowed determination of membrane permeability to potassium (8 × 10^?8 cm sec^?1) and membrane potassium conductance (25 μmho cm^?2). Furthermore, the use of tracer flux techniques has indicated that the exchangeable fraction of intracellular potassium is 204 ± 14 mM. This represents the bulk of egg potassium (222 ± 19 mM as determined from flame photometry). Studies of unidirectional potassium efflux have indicated that its movement out of the egg is made up of at least two components; an external potassium-independent potassium efflux and external potassium-dependent efflux, the latter possibly representing a potassium exchange mechanism. The combined electrophysiological and tracer-flux data indicate that only a small portion of the total membrane conductance is composed of potassium conductance at this stage of development. This and the fact that the membrane potential is far from the potassium equilibrium potential are similar to observations made on mature eggs of several other species.     

PERMEABILITY OF ERYTHROCYTES TO RADIOACTIVE POTASSIUM

The diffusion coefficients for the exchange of potassium across the membrane of erythrocytes of humans, rats, and rabbits have been determined by the use of artificially radioactive potassium, both into and out of the erythrocytes both in vitro and in vivo. The diffusion coefficients found in minutes^–1 were 0.2 to 0.25 x 10^–3 for human, 0.32 to 0.665 x 10^–3 for rabbits, and 1.0 x 10^–3 for rat erythrocytes. Rabbit erythrocytes appear to be more permeable in vivo. Reasons are advanced to explain the failure of earlier workers to demonstrate appreciable exchange of potassium in erythrocytes.     

Studies on some enzymes relevant to citric acid accumulation by Aspergillus niger

Enzymatic studies have been performed on a local strain of Aspergillus niger to find a correlation with citric acid accumulation. The activity of aconitase [aconitate hydratase, citrate(isocitrate) hydrolyase, EC 4.2.1.3] and isocitrate dehydrogenase (NADP⁺) [threo-d_s-isocitrate:NADP⁺ oxidoreductase (decarboxylating) EC 1.1.1.42] decreased after 4 days whereas that of citrate synthase [citrate oxaloacetate-lyase (pro-3S-CH₂COO^?→acetylCoA), EC 4.1.3.7] did so after 8 days, when citric acid accumulation in the medium reached a maximum (45.9 mg ml^?1). In vitro studies with mycelial cell-free extracts demonstrated inhibition of citrate synthase activity by sodium azide and potassium ferricyanide on both the 4th and 8th days. Aconitase was inhibited by sodium arsenate, sodium fluoride, iodoacetic acid and potassium ferricyanide only on the 4th day. Isocitrate dehydrogenase (NADP⁺) activity on the 4th and 8th days was inhibited by iodoacetic acid but was stimulated by potassium ferricyanide. The possible existence of isozyme species of these enzymes is discussed.     

Potassium and sodium transport across single distal tubules of Amphiuma

The transport properties of potassium (K) and sodium (Na) were studied in single distal tubules of Amphiuma using free-flow micropuncture techniques and stationary microperfusion methods. The transepithelial movement of labeled potassium was measured utilizing a three-compartment system in series in which the time course of tracer disappearance from the lumen was followed. Under control conditions, in blood- and doubly-perfused kidneys, extensive active net reabsorption of sodium and potassium obtains along single distal tubules. Tubular potassium reabsorption is abolished by ouabain at a concentration of 5 x 10^-6 M. Significant net secretion of K can be induced by exposing Amphiuma to a high K environment (100 mM KCl) or by adding acetazoleamide (1 x 10^-4 M) to the perfusion fluid. Transepithelial movement of potassium involves mixing with only a small fraction of total distal tubular cell potassium. This transport pool of potassium increases significantly with the transition from tubular net reabsorption to net secretion. Indirect evidence is presented which indicates that increased active K uptake across the peritubular cell boundary may be of prime importance during states of net K secretion.     

β-aminobutyric acid-mediated tobacco tolerance to potassium deficiency

??-Aminobutyric acid (BABA) is known as a nonprotein amino acid that can enhance the capacity of plants to resist various biotic and abiotic stresses. However, the role of BABA in protecting tobacco (Nicotiana tabacum L.) against potassium deficiency is not well understood. In this study, we demonstrated that the appropriate concentration of BABA promoted tobacco growth and enhanced plant tolerance to potassium deficiency. BABA protected tobacco from the reduction in the primary root length and chlorophyll content and increased K⁺ uptake via the regulation of expression of potassium up-taking-related NtHAK, NtLKS, NKT1, and NtKC genes. In addition, BABA affected the balancing between hydrogen peroxide level and peroxidase activity under low-potassium stress. Together, our results suggest that BABA plays a role in enhancing potassium deficiency stress tolerance by increasing K⁺ uptake, at least in part, via a ROS-dependent mechanism.     

Characterization of ATPase activity associated with corn leaf plasma membranes

A Mg²⁺-dependent, cation-stimulated ATPase was associated with plasma membranes isolated from corn leaf mesophyll protoplasts. Potassium was the preferred monovalent cation for stimulating the ATPase above the Mg²⁺-activated level. The enzyme was substrate-specific for ATP, was inhibited by N,N′-dicyclohexylcarbodiimide, diethylstilbestrol, p-chloromercuribenzoate, and orthovanadate, but was insensitive to oligomycin or sodium azide. A K_m of 0.28 millimolar Mg²⁺-ATP was determined for the K⁺-ATPase, and the principal effect of potassium was on the V_max for ATP hydrolysis. Since potassium stimulation was not saturated at high concentrations, a nonspecific role was proposed for potassium stimulation. A nonspecific phosphatase was also found to be associated with corn leaf plasma membranes. However, it could not be determined positively whether this activity represented a separate enzyme.     

Potassium conductances and proliferation in conditionally immortalized renal glomerular mesangial cells from the H-2Kb-tsA58 transgenic mouse

The effects of the temperature-sensitive, immortalizing Simian Virus 40 T antigen, tsA58, on whole-cell potassium conductances were assessed in renal glomerular mesangial cells from H-2K^b-tsA58 transgenic mice [1]. MTT cell viability assay data indicated that in permissive (33°C, 50 U ml⁻¹ γ-interferon, IFN+) and non-permissive (37°C, without γ-interferon, IFN−) culture conditions the oncogene was active and inactive respectively. In IFN+ cells whole-cell currents were inhibited by 10 mM 4-aminopyridine, 1 mM ATP and glibenclamide (glyburide, IC₅₀=0.4 μM) and stimulated by cromakalim (EC₅₀=40 μM). Furthermore, increases in pipette free calcium activity stimulated the potassium conductance (EC₅₀=0.5 μM). Apamin inhibited this conductance (IC₅₀=9 nM). None of these effects were observed in IFN− cells. The potassium conductance in IFN− cells was activated by a hyposmotic shock and this was inhibited by Gd³⁺. These data indicate that (1) conductances consistent with ATP-sensitive and small, calcium-activated potassium channels are found in IFN+ cells, (2) an osmotically-sensitive channel is found in IFN− cells and (3) channel expression is dependent upon the activation of tsA58.     

Osmotic adaptation of T4 infected Escherichia coli

In contrast to enzymatic adaptation, osmotic adaption is possible with T4-infected Escherichia coli B cells. After an osmotic shift from 220 mOsM to 690 mOsM the intracellular content of potassium rises in infected cells as well as in uninfected cells. After osmotic shock the involved TrKA transport system shows an increased discrimination against rubidium (Rb⁺) and for potassium (K⁺).     

Varietal differences in potassium uptake by barley

Potassium influx isotherms were obtained for 10 cultivars of barley using plants which had been grown with or without potassium (high K⁺ and low K⁺ plants, respectively), and the cultivars ranked with respect to K_m or V_max values for influx with a view to using these rankings as a predictive measure of long term performance under conditions of potassium-limited growth. Analyses of these rankings revealed significant differences between cultivars. Net uptake rates for low K⁺ plants, determined over a 24-hour period, confirmed the differences between upper (Herta) and lower (Conquest) ranked cultivars, and established similar differences in the rates of translocation to the shoot. Efflux analyses showed no differences in potassium efflux from the cytoplasm or from the vacuole for these cultivars. Growth rate studies under different conditions of potassium limitation indicated, with some exceptions, strong positive correlations between ranks accorded cultivars on the basis of influx kinetics and those based upon growth rates.     

The action of valinomycin in uncoupling corn mitochondria

Valinomycin in the presence of potassium is a potent uncoupler of corn (Zea mays L.) mitochondria, eliminating respiratory control. Valinomycin produces higher steady state potassium phosphate swelling which can be reversed to give active shrinkage if mersalyl is added to block the Pi⁻/OH⁻ antiporter. Respiration declines concurrently. Uncouplers accelerate the shrinkage and restore the respiration. The same results can be obtained with sodium phosphate if gramicidin D is substituted as ionophore.     

The kinetics of sodium extrusion in striated muscle as functions of the external sodium and potassium ion concentrations

After a 20 min initial washout, the rate of loss of radioactively labeled sodium ions from sodium-enriched muscle cells is sensitive to the external sodium and potassium ion concentrations. In the absence of external potassium ions, the presence of external sodium ions increases the sodium efflux. In the presence of external potassium ions, the presence of external sodium ions decreases the sodium efflux. In the absence of external potassium ions about one-third of the Na⁺ efflux that depends upon the external sodium ion concentration can be abolished by 10^-5 M glycoside. The glycoside-insensitive but external sodium-dependent Na⁺ efflux is uninfluenced by external potassium ions. In the absence of both external sodium and potassium ions the sodium efflux is relatively insensitive to the presence of 10^-5 M glycoside. The maximal external sodium-dependent sodium efflux in the absence of external potassium ions is about 20% of the magnitude of the maximal potassium-dependent sodium efflux. The magnitude of the glycoside-sensitive sodium efflux in K-free Ringer solution is less than 10% of that observed when sodium efflux is maximally activated by potassium ions. The inhibition of the potassium-activated sodium efflux by external sodium ions is of the competitive type. Reducing the external sodium ion concentration displaces the plots of sodium extrusion rate vs. [K]_o to the left and upwards.     

Expression, Purification and Functional Reconstitution of Slack Sodium-Activated Potassium Channels

The slack (slo2.2) gene codes for a potassium-channel α-subunit of the 6TM voltage-gated channel family. Expression of slack results in Na⁺-activated potassium channel activity in various cell types. We describe the purification and reconstitution of Slack protein and show that the Slack α-subunit alone is sufficient for potassium channel activity activated by sodium ions as assayed in planar bilayer membranes and in membrane vesicles.