Effect of Respiratory Inhibitors on the Motility of Pseudomonas fluorescens

The translational motility of Pseudomonas fluorescens was weakly inhibited by oligomycin, Dicumarol, 2,4-dinitrophenol, 2n-heptyl-4-hydroxyquinoline N-oxide, and potassium cyanide. Atabrine and antimycin A together with potassium cyanide immediately immobilized this bacterium, but antimycin A alone was without effect. Gramicidin D also immobilized P. fluorescens, but its action was inhibited by K(+) and NH(4) (+) ions. In like manner, the effect of p-chloromercuribenzoate could be counteracted with cysteine, thereby suggesting the involvement of -SH groups in flagellar motility processes. It appears that the energy required for motility of P. fluorescens is generated by oxidative phosphorylation mediated by the cytochrome system.     

Kaempferol inhibitions of corn mitochondrial phosphorylation

Kaempferol (3, 5, 7, 4′tetrahydroxyflavone) inhibited the rate of state 3 substrate oxidation, but not the state 4 rate. This, along with the kaempferol inhibition of substrate-driven calcium-phosphate deposition, provided evidence that kaempferol was acting specifically on the phosphorylation mechanism and not on electron transfer. Kaempferol, however, did not inhibit ATP-driven contraction while oligomycin did. Comparisons of kaempferol with mersalyl indicated that kaempferol did not inhibit phosphorylation by blocking phosphate transport. Both kaempferol and 2,4-dinitrophenol inhibited calcium-phosphate transport, but kaempferol did not stimulate respiration to the extent that 2,4-dinitrophenol did under acceptorless conditions. Kaempferol had no effect on NADH-driven contraction in a potassium chloride reaction medium. The site of kaempferol effect is thus seen to be unique from oligomycin and more like aurovertin, likely acting before the formation of the phosphorylated high energy intermediate, but not as an uncoupler in the traditional 2,4-dinitrophenol mode.     

Cell surface labeling of embryonic neural retina cells exposed to low temperature,energy inhibitors,cytochalasin B and colchicin

Large blebs devoid of receptors for hemocyanin-labeled concanavalin A appear on neural retina cells from 8-day chick embryos exposed to low temperature or to potassium cyanide or 2,4(α)-dinitrophenol at room temperature. Labeling with mixed antibodies against a crude retina membrane preparation and with goat anti-rabbit-hemocyanin conjugate showed the same results. Determination of cell ATP content indicated a drop in ATP concentration after exposure to low temperature or to respiration inhibitors. Disruption of microtubules by colchicin inhibited the formation of ‘naked’ large blebs, whereas cytochalasin B had no such inhibitory effect.     

Evidence for the extracellular reduction of alpha-lipoic acid by Leishmania donovani promastigotes: a transplasma membrane redox system

Leishmania donovani cells, capable of reducing certain electron acceptors with redox potentials at pH 7.0 down to -290 mV, outside the plasma membrane, can reduce the oxidised form of alpha-lipoic acid. alpha-Lipoic acid has been used as natural electron acceptor probe for studying the mechanism of transplasma membrane electron transport. Transmembrane alpha-lipoic acid reduction by Leishmania was not inhibited by mitochondrial inhibitors as azide, cyanide, rotenone or antimycin A, but responded to hemin, modifiers of sulphhydryl groups and inhibitor of glycolysis. The protonophores carbonyl cyanide chlorophenylhydrazone and 2,4-dinitrophenol showed inhibition of alpha-lipoic acid reduction. This transmembrane redox system differs from that of mammalian cells in respect to its sensitivity of UV irradiation and stimulation by diphenylamine. Thus a naphthoquinone coenzyme appears to be involved in alpha-lipoic acid reduction by Leishmania cells.     

THE UPTAKE OF SPERMIDINE AND SPERMINE BY SLICES OF MOUSE CEREBRAL HEMISPHERES

Abstract— Spermidine and spermine are taken up into mouse cerebral hemisphere slices by active transport and can be accumulated well above the medium concentration. The uptake process shows saturation kinetics and resembles that for amino acid uptake in that it is sensitive to temperature and inhibited by cyanide, 2,4-dinitrophenol or by the absence of glucose from the medium. However, at low initial medium concentrations spermine is taken up by a process which is insensitive to metabolic inhibitors or to temperature. It is suggested that either physical binding to a cellular constituent or exchange transport may account for this uptake. Ouabain does not inhibit polyamine uptake. Spermidine or spermine uptake is inhibited by cadaverine and putrescine. Spermine is the most potent inhibitor of spermidine uptake and vice-versa. Polyamine uptake differs from that of amino acids in that it is increased by a reduction in medium sodium or calcium content and decreased by an increase in medium potassium content. Recently taken up spermine undergoes heteroexchange with spermidine and homoexchange with recently entered spermine. Spermidine undergoes neither heteroexchange with spermine nor homoexchange.     

Biophysical properties and metabolic regulation of a TASK-like potassium channel in rat carotid body type 1 cells

The single channel properties of TASK-like oxygen-sensitive potassium channels were studied in rat carotid body type 1 cells. We observed channels with rapid bursting kinetics, active at resting membrane potentials. These channels were highly potassium selective with a slope conductance of 14-16 pS, values similar to those reported for TASK-1. In the absence of extracellular divalent cations, however, single channel conductance increased to 28 pS in a manner similar to that reported for TASK-3. After patch excision, channel activity ran down rapidly. Channel activity in inside-out patches was markedly increased by 2 and 5 mM ATP and by 2 mM ADP but not by 100 microM ADP or 1 mM AMP. In cell-attached patches, both cyanide and 2,4-dinitrophenol strongly inhibited channel activity. We conclude that 1) whilst the properties of this channel are consistent with it being a TASK-like potassium channel they do not precisely conform to those of either TASK-1 or TASK-3, 2) channel activity is highly dependent on cytosolic factors including ATP, and 3) changes in energy metabolism may play a role in regulating the activity of these background K+ channels.     

Alternate pathways of D-fructose transport and metabolism in Arthrobacter pyridinolis

Phosphoenolpyruvate: fructose phosphotransferase negative mutants of $\text{Arthrobacter}\text{pyridinolis}$ used fructose in the presence of malate. In isolated membrane vesicles from fructose-grown wild type cells, fructose uptake was stimulated by malate, but not by a variety of other compounds, to almost the same extent as by phosphoenolpyruvate. The stimulation by malate but not that by phosphoenolpyruvate was prevented by 2,4-dinitrophenol or potassium cyanide. D-Fructokinase activity was elevated in cells grown in malate plus fructose or sucrose.     

EXPERIMENTALLY INDUCED CHROMOSOME ABERRATIONS IN PLANTS : I. THE PRODUCTION OF CHROMOSOME ABERRATIONS BY CYANIDE AND OTHER HEAVY METAL COMPLEXING AGENTS

The finding of Lilly and Thoday that potassium cyanide produces structural chromosome changes in root tips of Vicia faba was confirmed. Like mustards, diepoxides, and maleic hydrazide, potassium cyanide seems to act on cells at early interphase. A tendency of cyanide breaks to be concentrated in heterochromatic segments of the chromosomes was evident. The production of chromosome aberrations by cyanide proved to be practically unaffected by the temperature during treatment. In agreement with Lilly and Thoday, the effect of potassium cyanide was found to be dependent on oxygen tension during treatment. The effect of potassium cyanide increases with increasing oxygen concentration up to 100 per cent oxygen. In the absence of oxygen, potassium cyanide was not completely inactive, but produced a low, though significant frequency of aberrations. Pretreatments with 2.4-dinitrophenol did not influence the effect of potassium cyanide. When bean roots were treated with potassium cyanide before a treatment with 8-ethoxycaffeine, or at the same time as they were treated with 8-ethoxycaffeine, the effect of 8-ethoxycaffeine was almost completely suppressed. The effects of a number of other heavy metal complexing agents were also tested. Sodium fluoride, potassium thiocyanate, carbon monoxide, o-phenanthroline, 2.2-bipyridine, and sodium azide were without radiomimetic effect under the conditions employed, and so was a mixture of sodium azide and sodium fluoride. A low, but quite significant, radiomimetic effect was obtained after treatments with sodium diethyldithiocarbamate, cupferron, and 8-hydroxyquinoline. Under anaerobic conditions, the effects of cyanide and cupferron were both quantitatively and qualitatively indistinguishable. Unlike the effect of cyanide, the effect of cupferron was not enhanced by the presence of oxygen. The effects of the same heavy metal complexing agents were tested on the activities of the enzymes catalase and peroxidase. The activities of both of these enzymes were found to be totally inhibited only by potassium cyanide. In the other cases, little correlation was found between ability to inhibit the activities of these enzymes and ability to produce chromosome aberrations. In a number of experiments, hydrogen peroxide was found to be without radiomimetic effect, whether alone or in combination with potassium cyanide. t-Butyl hydroperoxide proved to be active. The effect of t-butyl hydroperoxide was substantially increased by pretreatments with 2.4.-dinitrophenol. The results are discussed, and it is concluded that the observations made do not support the hypothesis that hydrogen peroxide is involved in the production of chromosome aberrations by potassium cyanide. The possibility that organic peroxides are involved cannot be excluded on the bases of the experimental results. As an alternative hypothesis, it is suggested that iron or other heavy metals are present in the chromosomes and that cyanide and other heavy metal complexing agents produce chromosome aberrations by reacting with these metals.     

Metabolic energy and cytoskeletal requirements for synthesis and secretion by acini from rat mammary gland—ii. Intracellular transport and secretion of protein and lactose

• 1.1. Iodoacetate, 2,4-dinitrophenol, cyanide and cycloheximide inhibited protein secretion as well as synthesis by acini (alveoli) from rat mammary gland. Cytochalasin B and vinblastine inhibited protein secretion and marginally reduced protein synthesis. Colchicine was without effect on protein synthesis but inhibited secretion.
• 2.2. Intracellular protein transport was altered during incubation with metabolic and cytoskeletal inhibitors. Cycloheximide, iodoacetate. 2,4-dinitrophenol and Cytochalasin B appeared to block protein synthesis on polysomes of rough endoplasmic reticulum. Vinblastine inhibited protein transport from rough endoplasmic reticulum to Golgi apparatus and colchicine appeared to cause accumulation of protein in several endomembrane fractions.
• 3.3. Iodoacetate reduced acinar lactose content but was without effect on lactose synthetase activity. Cyanide, cycloheximide and vinblastine reduced lactose synthetase activity but not tissue lactose concentration. Cytochalasin B reduced glucose incorporation but was without effect on lactose content and lactose synthetase activity. Colchicine and 2,4-dinitrophenol did not alter glucose incorporation, lactose content or lactose synthetase activity. Lactose secretion was inhibited by all metabolic and cytoskeletal inhibitors examined.
• 4.4. Results indicated that sustained protein secretion depended on continued protein synthesis and that lactose secretion was coupled to protein secretion.

     

Effects of toxic substances on natural bacterial assemblages determined by means of [h]thymidine incorporation

The effects of 3,5-dichlorophenol, 2,4-dinitrophenol, and potassium dichromate on natural bacterial assemblages were examined by means of [H]thymidine incorporation into trichloroacetic acid-insoluble material. Results from a large number of coastal marine and freshwater samples suggest the following. (i) The effects of the three toxicants included reductions in the bacterial cell number as well as changes in rates of [H]thymidine incorporation and in [H]thymidine incorporation per cell. The concentrations that inhibited [H]thymidine incorporation by 50% ranged from 3 to 11 mg liter for 3,5-dichlorophenol, 6 to 10 mg liter for 2,4-dinitrophenol, and 21 to 123 mg liter for potassium dichromate, with a tendency to higher values in bacterial assemblages from more eutrophic environments. (ii) The effects of 3,5-dichlorophenol and potassium dichromate determined by [H]leucine incorporation into bacterial protein were similar or larger than those obtained from [H]thymidine incorporation. (iii) Two to four hours of exposure to the toxicants was necessary before stable maximum effects were found in [H]thymidine incorporation. (iv) Storage of natural environmental samples should be avoided, since tests with water stored for 1 to 3 days sometimes produced results different from results obtained from in situ tests. (v) The effects of 3,5-dichlorophenol, 2,4-dinitrophenol, and potassium dichromate on natural bacterial assemblages were relatively constant during periods with different growth rates in the assemblages, during various periods of the year, and between samples from freshwater and marine localities. With some precautions, [H]thymidine incorporation can be used as a quick and sensitive method for determining the effects of toxicants on aquatic bacterial assemblages from natural environmental samples.     

The immediate uptake of potassium ion by mitochondria requiring gramicidin and 2,2-dimethylsuccinic anhydride.

The addition of 2,2-dimethylsuccinic anhydride to mitochondrial suspensions fortified with gramicidin and potassium ion but without any permeant anion caused an immediate and rapid increase in volume (as indicated by absorbance change at 520 nm) and the uptake of potassium ion (as indicated by a cation-specific electrode). The phenomena was not inhibited by rutamycin but was inhibited by either rotenone, antimycin or 2,4-dinitrophenol. Rotenone inhibition was relieved by succinate thus one of the requirements of the process was energy derived from endogenous substrates. Potassium ion could be replaced by rubidium and cesium ions but not by lithium or sodium ions. Since 2,2-dimethylsuccinate could not replace the anhydride and was not a permeant anion there must also be a requirement for the anhydride bond. The action of the anhydride on the mitochondria must be direct. Only closely related anhydrides were capable of engendering the effect of a highly effective permeant anion.     

Effect of xenobiotics on inorganic nitrogen assimilation by the phototrophic bacteriumRhodobacter capsulatus E1F1

In the presence of nitroaromatic and haloaromatic derivatives,Rhodobacter capsulatus E1F1 growth was affected in different degrees depending on the nitrogen source used. Phototrophic growth on glutamate or ammonium was not inhibited by 2,4-dinitrophenol (2,4-DNP), 4-nitrophenol (4-NP), 2-amino-4-nitrophenol (2,4-ANP), 4-aminophenol (4-AP), or 4-chlorophenol (4-CIP), whereas 2,4-dinitrophenol and 4-chlorophenol partially inhibited bacterial growth in nitrate, nitrite, and dinitrogen. On the other hand, although photosynthetic nitrate uptake was significantly inhibited by 2,4-dinitrophenol, 4-chlorophenol inhibited it to a lesser extent. Nitrogen fixation was severely inactivated in vivo by 2,4-dinitrophenol, but nitrate reductase activity was activated in vivo by 2,4-dinitrophenol, 4-nitrophenol, and 4-chlorophenol. Similar effects were found in cells growing with nitrate and 2,4-dinitrophenol under dark and aerobiosis conditions. None of the enzymatic activities related to inorganic nitrogen assimilation were affected by xenobiotics in vitro.     

Lysine and alanine transport in the perfused guinea-pig placenta

The characteristics of L-lysine transport were investigated at brush-border (maternal) and basal (fetal) sides of the syncytiotrophoblast in the term guinea-pig placenta artificially perfused either through the umbilical vessels in situ or through both circulations simultaneously. Cellular uptake, efflux and transplacental transfer were determined using a single-circulation paired-tracer dilution technique. Unidirectional L-[3H]lysine uptake (%) (perfusate lysine 50 microM) was high on maternal (M = 87 +/- 1) and fetal (F = 73 +/- 2) sides. L-[3H]Lysine efflux back into the ipsilateral circulation was asymmetrical (F/M ratio = 2.3) and transplacental flux occurred in favour of the fetal circulation. Unidirectional lysine influx kinetics (0.05-8.00 mM) gave Km values of 1.75 +/- 0.70 mM and 0.90 +/- 0.25 mM at maternal and fetal sides, respectively; corresponding Vmax values were 1.95 +/- 0.38 and 0.87 +/- 0.10 mumol.min-1.g-1. At both sides, lysine influx (50 microM) could be inhibited (about 60-80%) by 4 mM L-lysine and L-ornithine and less effectively (about 10-40%) by L-citrulline, L-arginine, D-lysine and L-histidine. At the basal side: (i) lysine influx kinetics were greatly modified in the presence of 10 mM L-alanine (Km = 6.25 +/- 3.27 mM; Vmax = 2.62 +/- 0.94 mumol.min-1.g-1), but unchanged by equimolar L-phenylalanine or L-tryptophan; (ii) in the converse experiments, lysine (10 mM) did not affect the kinetic characteristics for either L-alanine or L-phenylalanine; (iii) L-lysine and L-alanine influx kinetics were not dependent on the sodium gradient; (iv) the inhibition of L-[3H]lysine uptake by 4 mM L-homoserine was partially (60%) Na+-dependent. At the maternal side the kinetic characteristics for alanine influx were highly Na+-dependent, while lysine influx was partially Na+-dependent only at low concentrations (0.05-0.5 mM). Bilateral perfusion with 2,4-dinitrophenol (1 mM) reduced L-[3H]lysine uptake into the trophoblast and abolished transplacental transfer. It is suggested that lysine transport in the guinea-pig placenta is mediated by a specific transport system (y+) for cationic amino-acids. The asymmetry in the degree of sodium-dependency at both trophoblast membranes may in part explain the maternal-to-foetal polarity of placental amino-acid transfer in vivo.     

The Uptake of Carnitine by Slices of Rat Cerebral Cortex

Abstract: The properties of carnitine transport were studied in rat brain slices. A rapid uptake system for carnitine was observed, with tissue-medium gradients of 38 ± 3 for L-[¹⁴CH₃]carnitine and 27 ± 3 for D-[¹⁴CH₃]carnitine after 180 min incubation at 37°C in 0.64 mM substrate. Uptake of L- and D-carnitine showed saturability. The estimated values of K _m for L- and D-carnitine were 2.85 mM and 10.0 mM, respectively; but values of V _max (1 μmol/min/ml in-tracellular fluid) were the same for the two isomers. The transport system showed stereospecificity for L-carnitine. Carnitine uptake was inhibited by structurally related compounds with a four-carbon backbone containing a terminal carboxyl group. L-Carnitine uptake was competitively inhibited by γ-butyrobetaine ( K _i= 3.22 mM), acetylcarnitine ( K _i= 6.36 mM), and γ-aminobutyric acid ( K _i= 0.63 mM). The data suggest that carnitine and γ-aminobutyric acid interact at a common carrier site. Transport was not significantly reduced by choline or lysine. Carnitine uptake was inhibited by an N₂ atmosphere, 2,4-dinitrophenol, carbonylcyanide- N -chlorophenylhydrazone, potassium cyanide, n-ethylmaleimide, and ouabain. Transport was abolished by low temperature (4°C) and absence of glucose from the medium. Carnitine uptake was Na⁺-dependent, but did not require K⁺ or Ca²⁺.     

Experimentally induced chromosome aberrations in plants. I. The production of chromosome aberrations by cyanide and other heavy metal complexing agents

The finding of Lilly and Thoday that potassium cyanide produces structural chromosome changes in root tips of Vicia faba was confirmed. Like mustards, diepoxides, and maleic hydrazide, potassium cyanide seems to act on cells at early interphase. A tendency of cyanide breaks to be concentrated in heterochromatic segments of the chromosomes was evident. The production of chromosome aberrations by cyanide proved to be practically unaffected by the temperature during treatment. In agreement with Lilly and Thoday, the effect of potassium cyanide was found to be dependent on oxygen tension during treatment. The effect of potassium cyanide increases with increasing oxygen concentration up to 100 per cent oxygen. In the absence of oxygen, potassium cyanide was not completely inactive, but produced a low, though significant frequency of aberrations. Pretreatments with 2.4-dinitrophenol did not influence the effect of potassium cyanide. When bean roots were treated with potassium cyanide before a treatment with 8-ethoxycaffeine, or at the same time as they were treated with 8-ethoxycaffeine, the effect of 8-ethoxycaffeine was almost completely suppressed. The effects of a number of other heavy metal complexing agents were also tested. Sodium fluoride, potassium thiocyanate, carbon monoxide, o-phenanthroline, 2.2-bipyridine, and sodium azide were without radiomimetic effect under the conditions employed, and so was a mixture of sodium azide and sodium fluoride. A low, but quite significant, radiomimetic effect was obtained after treatments with sodium diethyldithiocarbamate, cupferron, and 8-hydroxyquinoline. Under anaerobic conditions, the effects of cyanide and cupferron were both quantitatively and qualitatively indistinguishable. Unlike the effect of cyanide, the effect of cupferron was not enhanced by the presence of oxygen. The effects of the same heavy metal complexing agents were tested on the activities of the enzymes catalase and peroxidase. The activities of both of these enzymes were found to be totally inhibited only by potassium cyanide. In the other cases, little correlation was found between ability to inhibit the activities of these enzymes and ability to produce chromosome aberrations. In a number of experiments, hydrogen peroxide was found to be without radiomimetic effect, whether alone or in combination with potassium cyanide. t-Butyl hydroperoxide proved to be active. The effect of t-butyl hydroperoxide was substantially increased by pretreatments with 2.4.-dinitrophenol. The results are discussed, and it is concluded that the observations made do not support the hypothesis that hydrogen peroxide is involved in the production of chromosome aberrations by potassium cyanide. The possibility that organic peroxides are involved cannot be excluded on the bases of the experimental results. As an alternative hypothesis, it is suggested that iron or other heavy metals are present in the chromosomes and that cyanide and other heavy metal complexing agents produce chromosome aberrations by reacting with these metals.     

Multiphasic Uptake of Amino Acids by Barley Roots

Concentration-dependence and other characteristics of uptake of ³H-labeled l -lysine, l -methionine and l -proline by excised roots of barley (Hordeum vulgare L.) were studied. Use of relatively short uptake and wash periods and low solute concentrations ensured good estimates of influx across the plasmalemma. Uptake in the range of 10^?7M– 6.3 × 10^?3M can be precisely represented by four or five phases of single, multiphasic mechanisms. The mechanisms appear to be relatively specific as judged from the competition by unlabeled analogues. Structural requirements for interaction of a compound with the uptake site for methionine are given, as are the effects of analogues on the phase pattern for this amino acid. There is no indication of separate uptake and transition sites for methionine or lysine. i.e. phase transitions seem in this case to be caused by binding of molecule(s) to the uptake site. Uptake, but not phase patterns, was highly pH-dependent. The optima were pH 5 for lysine, pH 3–5 (a broad peak) for methionine and about pH 5.5 for proline. Uptake of the three amino acids was strongly inhibited by 2,4-dinitrophenol. sulfhydryl reagents and deoxycholate.     

Studies on the sodium and potassium transport in rabbit polymorphonuclear leukocytes

Rabbit polymorphonuclear leukocytes obtained from peritoneal exudates, incubated at 37°C. following exposure to 4°C., actively reaccumulate potassium while little or no net extrusion of sodium takes place. Preventing the utilization of oxidative metabolism with potassium cyanide, 2,4-dinitrophenol, or a nitrogen atmosphere does not inhibit the recovery process. Inhibitors blocking anaerobic glycolysis (sodium iodoacetate and sodium fluoride in low concentrations) completely abolish the capacity to reaccumulate potassium and cause a further dissipation of the sodium and potassium gradients. Water movements have been shown to be secondary to cation shifts. It is postulated that separate transport mechanisms exist for sodium and potassium and that the process of potassium reaccumulation relies on anaerobic glycolysis as a source of energy.     

Effect of 2,4-Dinitrophenol on Auxin-induced Ethylene Production and Auxin Conjugation by Mung Bean Tissue

Auxin-induced ethylene production by mung bean (Phaseolus mungo L.) hypocotyl segments was markedly inhibited by 2,4-dinitrophenol regardless of whether or not kinetin was present. Uptake of indoleacetic acid-2-¹⁴C was also inhibited in the presence of 2,4-dinitrophenol. Segments treated only with indoleacetic acid rapidly converted indoleacetic acid into indole-3-acetylaspartic acid with time whereas kinetin suppressed indoleacetic acid conjugation. Formation of indole-3-acetylaspartic acid was significantly reduced when 2,4-dinitrophenol was present. The suppression of indoleacetic acid conjugation by kinetin and 2,4-dinitrophenol appeared to be additive, and the free indoleacetic acid level in segments treated with 2,4-dinitrophenol in the presence of indoleacetic acid or indoleacetic acid plus kinetin was remarkably higher than in corresponding segments which received no 2,4-dinitrophenol.     

Dependency of the ATPase and 32 P--ATP exchange reaction of mitochondria on K + and electron transport

The 2,4-dinitrophenol-stimulated ATPase activity and the ³²P-ATP exchange reaction has been studied in rat liver mitochondria having less than 15 nmoles of K⁺ per milligram of protein. With 200 mm sucrose in the incubation media, the permeation of K⁺ and an oxidizable substrate is required for maximal stimulation of ATPase activity by 2,4-dinitrophenol. In these conditions, the 2,4-dinitrophenol-stimulated ATPase is inhibited by antimycin, acetate and mersalyl and depends to a certain extent on the rate of electron transport. The ³²P-ATP exchange reaction of mitochondria with a low content of K⁺ also requires K⁺ permeation and is inhibited by antimycin, cyanide, 2,4-dinitrophenol, and acetate. The results suggest that the entrance of ATP into the mitochondria is compulsory linked to K⁺ uptake in a process that depends on a negative internal potential.     

Investigation of respiratory and ion transport properties of aging bean stem slices

Ion and oxygen uptake were studied on aging bean stem slices. Oxygen uptake was high immediately after slicing, decreased to a minimum at 100 minutes, and then increased again. Ion uptake per unit of O₂ uptake data suggested that metabolic energy was utilized almost exclusively for sodium transport in fresh tissue but was diverted to potassium transport as the slices aged. Oxygen and ion uptake in fresh slices was less sensitive to 2,4-dinitrophenol as compared to the aged slices, indicating major metabolic and physiological changes occurred during aging. This was further substantiated by the tissue response to cyanide and antimycin A. Oxygen uptake was decreased by cyanide (22% by 1 mm) and antimycin A (14% by 1 microgram per milliliter) in fresh slices but not in aged slices. Potassium uptake that developed during aging was sensitive to cyanide and antimycin A. The results are pertinent to understanding the role of the stem in regulating ion transport in plants.