Effect of potassium ions at the different concentration on the interaction between AmB and the lipid monolayer containing cholesterol or ergosterol

AmB is an antifungal drug of polyene. Although it is prone to nephrotoxicity, it is still the gold standard in the clinical treatment of fungal infection. Sterol plays a decisive role in the drug activity of AmB. The antifungal activity of AmB depends on ergosterol in fungal membranes, and its toxicity is related to cholesterol in mammalian membranes. At the same time, AmB interacts with biofilms, leading to a significant loss of potassium ions and affecting the transport of potassium ions across membranes. Meanwhile, metal cation may also affect AmB molecules’ aggregation on the membrane. This paper mainly studied the effects of different concentrations of potassium ions on the interactions between AmB and lipid monolayers containing cholesterol or ergosterol and explored the differences in the impact of varying potassium ions on the drug activity of AmB on monolayers rich in these two kinds of sterols. The results show that potassium ions caused the collapse of lipid monolayer and lipid-AmB monolayer to disappear. The limiting molecular area of these monolayers also increased due to potassium ions. The limiting molecular area of the monolayer in the presence of ergosterol has a great difference in the different concentration of potassium ions, which is different from that in the presence of cholesterol. The presence of potassium ions, regardless of the intensity of K⁺ ions, increased the maximum elastic modulus of the lipid/sterol monolayer with and without AmB. The presence of potassium ions reduced the influence of AmB on the stability of the lipid monolayer containing cholesterol. The impact of AmB on the stability of the lipid monolayer containing ergosterol was related to the concentration of potassium ions. The potassium ions increased the area of the ordered “island” region on the lipid-AmB monolayer containing cholesterol, and the boundary of the microregion produced different degrees of curvature. However, on the lipid/ergosterol monolayer, 5 mM and 10 mM potassium ions made the holes caused by AmB more denser, and the diameter of holes become larger. These results can help to improve the effect of potassium ions on the transmembrane transport of substances affected by AmB. The results will provide a basis for further exploration of the effect mechanism of metal ions on the antifungal activity of polyene drugs.     

Flexibility of an Active Center in Sodium-Plus-Potassium Adenosine Triphosphatase

In plasma membranes of intact cells an enzymatic pump actively transports sodium ions inward and potassium ions outward. In preparations of broken membranes it appears as an adenosine triphosphatase dependent on magnesium, sodium, and potassium ions together. In this adenosine triphosphatase a phosphorylated intermediate is formed from adenosine triphosphate in the presence of sodium ions and is hydrolyzed with the addition of potassium ions. The normal intermediate was not split by adenosine diphosphate. However, selective poisoning by N-ethylmaleimide or partial inhibition by a low magnesium ion concentration yielded an intermediate split by adenosine diphosphate and insensitive to potassium ions. Pulse experiments on the native enzyme supported further a hypothesis of a sequence of phosphorylated forms, the first being made reversibly from adenosine triphosphate in the presence of sodium ion and the second being made irreversiblyfrom the first and hydrolyzed in the presence of potassium ion. The cardioactive steriod inhibitor, ouabain, appeared to combine preferentially with the second form. Phosphorylation was at the same active site according to electrophoretic patterns of proteolytic phosphorylated fragments of both reactive forms. It is concluded that there is a conformational change in the active center for phosphorylation during the normal reaction sequence. This change may be linked to one required theoretically for active translocation of ions across the cell membrane.     

Net uptake of potassium in Neurospora. Exchange for sodium and hydrogen ions

Net uptake of potassium by low K, high Na cells of Neurospora at pH 5.8 is accompanied by net extrusion of sodium and hydrogen ions. The amount of potassium taken up by the cells is matched by the sum of sodium and hydrogen ions lost, under a variety of conditions: prolonged preincubation, partial respiratory inhibition (DNP), and lowered [K]_o. All three fluxes are exponential with time and obey Michaelis kinetics as functions of [K]_o. The V _max for net potassium uptake, 22.7 mmoles/kg cell water/min, is very close to that for K/K exchange reported previously (20 mmoles/kg cell water/min). However, the apparent K_m for net potassium uptake, 11.8 mM [K]_o, is an order of magnitude larger than the value (1 mM) for K/K exchange. It is suggested that a single transport system handles both net K uptake and K/K exchange, but that the affinity of the external site for potassium is influenced by the species of ion being extruded.     

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.     

The influence of potassium- and sodium-free solutions on sodium efflux from squid giant axons

The sensitivity of sodium efflux to the removal of potassium ions from the external solution and the change in sodium efflux occurring when sodium ions are also removed were observed to be related. When Tris was used to replace external sodium ions, increases in sodium efflux were always observed whether the sensitivity of sodium efflux to external potassium ions was weak or strong. Greater percentage increases in sodium efflux occurred, however, the greater the sensitivity of sodium efflux to external potassium ions. When lithium ions were used to replace external sodium ions, increases in sodium efflux occurred if the sensitivity of efflux to external potassium ions was strong whereas decreases in sodium efflux took place if the sensitivity of efflux to external potassium ions was weak. Intermediate sensitivities of efflux to external potassium resulted in no change in efflux upon substitution of lithium ions for external sodium ions. In the presence of 10^-5 M ouabain, substitution of Tris for external sodium ions always resulted in a small decrease in sodium efflux. The data can be described in terms of a model which assumes the presence of efflux stimulation sites that are about 98% selective to potassium ions and about 2% selective to sodium or lithium ions.     

Dual mechanism for stimulation of glutamate transport by potassium ions in Streptococcus mutans.

An ATP-driven primary transport system operative for L-glutamate or L-aspartate in Streptococcus mutans is, through the entire pH range from 5.5 to 8.5, specifically stimulated by extracellular potassium ions. The stimulation by potassium ions observed in the low pH range between 5.5 and 7 has been interpreted to be due to potassium ion-dependent regulation of the intracellular pH (the first mechanism). In the high pH range from 7 to 8.5, on the other hand, the present study demonstrates that potassium stimulation is essentially not associated with such intracellular pH regulation. This conclusion is based on our observation that potassium stimulation in the high pH range is insensitive to a proton conductor, carbonyl cyanide-p-trifluoromethoxy-phenyl-hydrazone. Since none of the other monovalent cations, including sodium, rubidium, ammonium, and Tris ions, could replace potassium ions in significantly stimulating glutamate transport, it is most likely that the influx of potassium ions specifically cancels the membrane potential derived by movement of glutamate with the net negative charges across a membrane and thus facilitates transport (the second mechanism). The second mechanism appears to be operative even in a low pH range, in addition to the first mechanism.     

An electrophysiological study of the effects of ionophore A23187 on Nauphoeta salivary glands

The ionophore A23187 produces a hyperpolarization of cockroach salivary gland cells in the presence or absence of added calcium ions. The effect is greater and more prolonged in the presence of calcium and is dependent on the external potassium ion concentration. It is proposed that the ionophore can increase the intracellular calcium ion concentration by an increase in influx from the external medium or by mobilization of intracellular calcium stores and that this results in an increase in the potassim permeability, thus producing a hyperpolarization.     

Investigation of the TEA-resistant outward current in the somatic membrane of perfused nerve cells

Outward currents remaining after addition of 20–50 mM of tetraethylammonium (TEA) ions to the extracellular or intracellular solution, were investigated in perfused isolatedHelix neurons. After this addition, the inactivated inward current carried by potassium ions, the potential-dependent and kinetic characteristics of which differ from those of potassium outward currents suppressed by TEA, is preserved in the membrane. A component dependent on the inward calcium current was found in this TEA-resistant outward current; it was abolished by replacement of the extra-cellular calcium ions by magnesium ions, by blocking of the calcium channels by extracellular cadmium ions, and by their destruction by intracellular fluoride ions. Increasing the intracellular concentration of free calcium ions by perfusing the cell with solutions containing calcium-EGTA buffer potentiated the TEA-resistant component of the outward current, whereas removal of these ions with EGTA weakened it. It is concluded that a system of outward current channels whose activation depends on the presence of calcium ions near the inner surface of the membrane is present in the somatic membrane. It is suggested that to keep these channels capable of being activated, calcium ions must bind with the structures forming their internal opening.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 11, No. 5, pp. 460–468, September–October, 1979.     

Parallel pathways of potassium transport in the alga Hydrodictyon reticulatum. Effects of calcium

Inflow of potassium ions into the alga Hydrodictyon reticulatum is reduced in the dark, the reduction being accompanied by a change in the selectivity pattern with respect to alkali metal ions, observed in competition experiments and evaluated by the gnostic analysis as described by Kovanic. This suggests that in the light a special mechanism of potassium uptake with a characteristic selectivity is switched on. This mechanism can be also suppressed by too high (2 mmol/l) or too low (EGTA) concentration of calcium ions in the medium. Since the same applies to the light-induced alkalinization of the algal surroundings it seems that the light-induced potassium uptake is related to the light-induced alkalinization, e.g., via exchange of external potassium cations for intracellular protons.     

Changes in the Membrane Permeability of Frog's Sartorius Muscle Fibers in Ca-Free EDTA Solution

The changes in the membrane permeability to sodium, potassium, and chloride ions as well as the changes in the intracellular concentration of these ions were studied on frog sartorius muscles in Ca-free EDTA solution. It was found that the rate constants for potassium and chloride efflux became almost constant within 10 minutes in the absence of external calcium ions, that for potassium increasing to 1.5 to 2 times normal and that for chloride decreasing about one-half. The sodium influx in Ca-free EDTA solution, between 30 and 40 minutes, was about 4 times that in Ringer's solution. The intracellular sodium and potassium contents did not change appreciably but the intracellular chloride content had increased to about 4 times normal after 40 minutes. By applying the constant field theory to these results, it was concluded that (a) P_Cl did not change appreciably whereas P_K decreased to a level that, in the interval between 10 and 40 minutes, was about one-half normal, (b) P_Na increased until between 30 and 40 minutes it was about 8 times normal. The low value of the membrane potential between 30 and 40 minutes was explained in terms of the changes in the membrane permeability and the intracellular ion concentrations. The mechanism for membrane depolarization in this solution was briefly discussed.     

Ion mechanisms of hyperpolarizing afterpotentials accompanying epileptic discharges in neocortical neurons

Hyperpolarizing afterpotentials of penicillin-induced (local application) paroxysmal depolarizing shifts (PDS) in neurons of the sensorimotor cortex of the cat were studied. The pattern of membrane conductance changes within different segments of hyperpolarization and the data on the role of various ion currents in its generation allow us to conclude that hyperpolarizing afterpotentials accompanying PDS are of a composite nature and include the following components: (i) the initial component provided by an increased membrane permeability to chloride ions (presumably a synaptic GABA_A response); (ii) the second component resulting predominantly from a potassium current and representing presumably a GABA_B response; and (iii) the final component comprising mainly a calcium-activated potassium current. These components are present in all neurons, are not clearly demarcated as separate waves, and partially overlap with each other, thus forming a prolonged hyperpolarizing deflection of the potential.     

枣皮红色素的提取及其稳定性研究

对枣皮红色素的提取工艺及其稳定性进行了系统研究。结果表明枣皮红色素水溶性较好,色素溶液对光、热比较稳定;在酸、碱、金属离子、氧化剂、蔗糖等添加剂中具有较好的稳定性;但在含氧酸根阴离子、还原剂、Vc、苯甲酸钠、山梨酸钾等添加剂中稳定性较差。     

Membrane currents induced by inflow of sodium ions into mollusk giant neurons

Membrane currents induced by inflow of sodium ions were investigated in giant neurons of the molluskHelix pomatia during tetanic stimulation or prolonged membrane depolarization under voltage clamp conditions. The membrane current thus produced consists of two components, a fast component with a reversal potential close to the potassium equilibrium potential, and a slow component only slightly dependent on membrane potential in the region from −50 to −90 mV. Addition of strophanthin K to the external solution, or replacement of sodium in the external solution by lithium or calcium abolished the slow component of the membrane current and reduced the fast component. It is concluded that the slow component appears as the result of activation of the sodium pump under electrogenic conditions, where-as the fast component arises as the result of an increase in potassium permeability, possibly coupled with intensive activity of this pump.     

Mechanisms of ammonia and ammonium ion toxicity in animal cells: Transport across cell membranes

A model for transport of ammonia and ammonium ions across cell membranes is presented. The model suggests that ammonium ions compete with potassium ions for inward transport, over the cytoplasmic membrane, via potassium transport proteins like the Na⁺/K⁺-ATPase and the Na⁺K⁺2Cl⁻-cotransporter. It also explains the difference between the ammonia/ammonium that is added to the cells and which is formed by the cells during metabolism of amino acids, especially glutamine and glutamate. The ammonium transport and subsequent events lead to predictable intracellular and extracellular pH (pH_e) changes. Experiments which verified the model and the predicted consequences were performed by measurements of the pH_e in concentrated cell suspensions. Addition of ammonium ions caused a time-dependent pH_e increase which was inhibited by potassium ions. The test system is not per se specific for transport measurements but the effect of potassium ions on the pH_e strongly favors our suggested model. Simple diffusion of ammonium ions would not be counteracted by potassium ions. The results show that ammonium ion transport in the murine myeloma cell line (Sp2/0-Ag14) used is inhibited by an excess of potassium ions. Results from experiments with specific inhibitors of suggested transport proteins were not conclusive. It is postulated that one important toxic effect of ammonia/ammonium is an increased demand for maintenance energy, caused by the need to maintain ion gradients over the cytoplasmic membrane. The results also suggest that potassium ions can be used to detoxify ammonia/ammonium in animal cell cultivations.     

Effects of extracellular potassium ions on outward potassium current dependent on extracellular calcium ions

Changes in outward potassium current occurring in response to changes in the concentration of potassium ions in the extracellular medium were investigated in unidentified neurons isolated fromHelix pomatia using an intracellular perfusion technique. It was found that introducing potassium ions (5–10 mM) into the extracellular solution produces a reversible increase in the component of outward potassium current which is dependent on extracellular calcium ions. Increased amplitude of this component occurs as a result of attenuated inactivation of the current under the action of extracellular potassium.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 19, No. 3, pp. 351–356, May–June, 1987.     

Potassium and sodium ions in a potassium channel studied by molecular dynamics simulations

We have performed simulations of both a single potassium ion and a single sodium ion within the pore of the bacterial potassium channel KcsA. For both ions there is a dehydration energy barrier at the cytoplasmic mouth suggesting that the crystal structure is a closed conformation of the channel. There is a potential energy barrier for a sodium ion in the selectivity filter that is not seen for potassium. Radial distribution functions for both ions with the carbonyl oxygens of the selectivity filter indicate that sodium may interact more tightly with the filter than does potassium. This suggests that the key to the ion selectivity of KcsA is the greater dehydration energy of Na⁺ ions, and helps to explain the block of KcsA by internal Na⁺ ions.     

A study of the effects of flocalin on respiration and potassium transport of rat-heart and liver mitochondria

The effects of the drug flocalin, which possesses cardioprotective properties, on the respiration rates of rat-heart and liver mitochondria in different functional states, the efficiency of oxidative phosphorylation, as well as the transport of potassium ions in these organelles, were studied. It was found that flocalin at concentrations of 7–30 μm stimulated respiration of rat-heart and liver mitochondria in V ₂ and V ₄ states in the presence of succinic add as a respiration substrate in a potassium-containing medium. In the absence of potassium ions in the incubation medium, flocalin had no effect on mitochondrial respiration in these states. Studying the functioning of the potassium transport system revealed that flocalin at these concentrations dose-dependently activated the ATP-dependent transport of potassium ions in rat-heart and liver mitochondria. The data we obtained indicate that the cardioprotective effect of flocalin can be associated with activation of the ATP-dependent potassium channel of the inner mitochondrial membrane.     

Effects of cloacin DF13 on the functioning of the cytoplasmic membrane

Adsorption of cloacin DF13 onto sensitive cells induces a leakage of potassium ions after a lag of 15–20 min. The rate of the potassium efflux is proportional to the bacteriocin concentration used. The extent of the potassium loss depends on the concentrations of bacteriocin and of extracellular potassium. Leakage of potassium ions is accompanied by an uptake of sodium ions. Cloacin DF13 does not affect the transport of magnesium ions, phosphate ions or amino acids. About 10 min after addition of the bacteriocin the ATP content of the cells begins to decrease gradually to about 25% of the original level. Anaerobiosis, lack of energy source and inhibition of oxidative phosphorylation may protect the cells if applied within 10 min after addition of the bacteriocin. The results suggest that the action of cloacin DF13 proceeds through at least two distinct phases. Phase I is a period after bacteriocin adsorption which is reversible and in some way depends on oxidative phosphorylation. This phase does not cause any cellular damage. Phase II is irreversible and results in a disturbance of several cellular functions.     

The Effect of pH and Bicarbonate Ions on the Uptake of Salts by Disks of Red Beet

1. The uptake of potassium chloride by disks of beetroot in solutionsat different pH values has been followed over periods of severaldays.
2. It was found that, as the pH was raised from approximately6to 9 the rate of uptake of potassium from 0.0024 M. potassiumchloride increased, and there was a corresponding increase inthe amount of potassium held by the disks at satura tion. Boththe rate of uptake and the final content of potassium in diskskept in solutions at pH 8.5 were frequently double those atpH 6.
3. The rate of uptake of chloride ions was only slightlyaffectedby the pH value of the medium. The final chloride contentwasapproximately the same in disks kept at different pH values.
4. The above ‘pH effect’ on potassium uptake hasbeenshown to be related to the uptake of bicarbonate ions presentin the alkaline solution; the additional potassium ions accumulatedaccompanying the bicarbonate anions taken up.
5. The bicarbonateions accumulated are converted primarily tomalic acid.

     

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⁺.