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Query: UMLS:C0027960 (mole)
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1. Bicarbonate transport across human red cell membranes was studied between 0 and 10 degrees C at alkaline pH values by determining the efflux of 14C-labelled bicarbonate from resealed erythrocyte ghosts. Transfer of labelled CO2 was eliminated as a source of error, when formation of intracellular 14CO2 was inhibited with carbonic anhydrase inhibitors. The study showed that there are no fundamental differences between the characteristics of bicarbonate and of chloride self-exchange as has been inferred from previous studies of chloride-bicarbonate exchange. 2. Efflux of radioactivity could be reduced more than 99% by reversible and irreversible inhibitors of anion transport. Inhibition of both chloride and bicarbonate self-exchange was linearly related to the binding of 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS) to the membranes. Complete (i.e. greater than 99%) inhibition was obtained after binding of 1.2 x 10(6) DIDS molecules per cell. 3. Bicarbonate self-exchange proved a saturable function of bicarbonate concentration, with a maximum at external and internal concentrations of approximately 100 mM, showing self-depression at higher bicarbonate concentrations, and half-maximum exchange flux at a concentration of 10 mM. The results were consistent with the hypothesis that the exchange mechanism has two anion binding sites, one mediating ion transport and the other causing transport inhibition. 4. Maximum exchange flux of bicarbonate was about 30% larger thant that of chloride, and the affinity of bicarbonate for the transport site was about three times larger than that of chloride. The apparent activation energy of bicarbonate exchange was 28 kcal/mole, the same order of magnitude as found for other inorganic anions between 0 and 10 degrees C. 5. The ability of other inorganic anions to exchange with bicarbonate decreased in the sequence Cl greater than NO3 greater than F greater than Br greater than or equal to I, corresponding to the sequence of the rate of self-exchange of halides. 6. Counter-transport of bicarbonate could be driven by a chloride gradient, when ghosts containing KCl were suspended in a medium containing traces of labelled bicarbonate in addition to a non-permeating anion. Concentration ratios (ci/co) up to about 1000 could be obtained. 7. It is concluded that bicarbonate is transported by the inorganic anion exchange mechanism of the erythrocyte membrane. The slight differences between the exchange kinetics of chloride and bicarbonate were explained by differing affinities of the two anions for the two anion binding sites of the transport system.
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PMID:Bicarbonate exchange through the human red cell membrane determined with [14C] bicarbonate. 51 56

1. The steady-state tracer exchange flux of chloride was measured at 10-150 mM external chloride concentration, substituting either lactate or sucrose for chloride. The chloride flux saturates in both cases with a K 1/2 about 50 and 15 mM, respectively. 2. The inhibitory effect of other monovalent anions on the chloride transport was investigated by measuring the 36Cl- efflux into media where either bromide, nitrate, or thiocyanate had been substituted for part of the chloride. The sequence of increasing affinity for the chloride transport system was found to be: Br- less than Cl- less than SCN- = NO3-. 3. The chloride steady-state exchange flux in the presence of nitrate can be described by Michaelis-Menten kinetics with nitrate as a competitive inhibitor of the chloride flux. 4. The apparent activation energy (EA) was determined to be 67 +/- 6.2 kJ/mole, and was constant between 7 and 38 degrees C. 5. The membrane potential (Vm) was measured as a function of the concentration of external K+, substituting K+ for Na+. The transference number of K+ (tK) was estimated from the slope of Vm vs. log10 (K+)e, and tCl and tNa were calculated, neglecting current carried by ions other than Cl-, K+, and Na+. The diffusional net flux of K+ was calculated from the steady-state exchange flux of 42K+, assuming the flux ratio equation to be valid. From this value the K+ conductance and the Na+ and Cl- conductances were calculated. The experiments showed that GCl, GNa, and GK are all about 14 muS/cm2. 6. The net (conductive) chloride permeability derived from the chloride conductance was 4 x 10(-8) cm/sec compared with the apparent permeability of 6 x 10(-7) cm/sec as calculated from the chloride tracer exchange flux. These data suggest that about 95% of the chloride transport is mediated by an electrically silent exchange diffusion. 7. Comparable effects of phloretin (0.25 mM) on the net (conductive) permeability and the apparent permeability to chloride (about 80% inhibition) may indicate that the chloride exchange and conductance pathways are not completely separate and distinct modes of transport, but may involve common elements. The reduced chloride permeability in the presence of phloretin is estimated to be two orders of magnitude larger than the ground permeability of the cell membrane.
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PMID:Membrane potential, chloride exchange, and chloride conductance in Ehrlich mouse ascites tumour cells. 52 33

Single channel currents though apical membrane Cl channels of the secretory epithelial cell line T84 were measured to determine the anionic selectivity and concentration dependence of permeation. The current-voltage relation was rectified with single channel conductance increasing at positive potentials. At 0 mV the single channel conductance was 41 +/- 2 pS. Permeability, determined from reversal potentials, was optimal for anions with diameters between 0.4 and 0.5 nm. Anions of larger diameter had low permeability, consistent with a minimum pore diameter of 0.55 nm. Permeability for anions of similar size was largest for those ions with a more symmetrical charge distribution. Both HCO3 and H2PO4 had lower permeability than the similar-sized symmetrical anions, NO3 and ClO4. The permeability sequence was SCN greater than I approximately NO3 approximately ClO4 greater than Br greater than Cl greater than PF6 greater than HCO3 approximately F much greater than H2PO4. Highly permeant anions had lower relative single channel conductance, consistent with longer times of residence in the channel for these ions. The conductance sequence for anion efflux was NO3 greater than SCN approximately ClO4 greater than Cl approximately I approximately Br greater than PF6 greater than F approximately HCO3 much greater than H2PO4. At high internal concentrations, anions with low permeability and conductance reduced Cl influx consistent with block of the pore. The dependence of current on Cl concentration indicated that Cl can also occupy the channel long enough to limit current flow. Interaction of Cl and SCN within the conduction pathway is supported by the presence of a minimum in the conductance vs. mole fraction relation. These results indicate that this 40-pS Cl channel behaves as a multi-ion pathway in which other permeant anions could alter Cl flow across the apical membrane.
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PMID:Anion permeation in an apical membrane chloride channel of a secretory epithelial cell. 137 74

This paper provides the results of studies which characterized conductive 36Cl- flux in basolaterally enriched membrane vesicles prepared from rabbit renal outer medulla. Conductive 36Cl- uptake was studied under two different experimental conditions. In the first, 36Cl- flux was driven by an inside positive voltage created with oppositely directed Cl- and gluconate gradients. In the second, an inwardly direct K+ gradient was used to drive 36Cl- uptake. By these two methods, voltage-sensitive 36Cl- uptake was shown to comprise about 45 and 65%, respectively, of the initial rates of total 36Cl- flux. Separate paired studies demonstrated that the conductive 36Cl- uptake was inhibited by the Cl- channel blocker diphenylamine-2-carboxylate (DPC) with an IC50 for DPC of 154 microM. The voltage-dependent 36Cl- uptake had an activation energy of 6.4 kcal/mole. This 36Cl- conductance had an anion selectivity sequence of I- greater than Cl- greater than or equal to NO3- much greater than gluconate.
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PMID:Cl- transport in basolateral renal medullary vesicles: I. Cl- transport in intact vesicles. 230 71

1. Multiple conductance level ion channels were recorded in excised and cell-attached patches from cells of a mouse B lymphocyte hybridoma line. The reversal potential for the single-channel current was unaffected by the species of cation on the cytoplasmic face of the patch, but changed as the Cl- concentration was altered, indicating that the channel is anion selective. 2. The permeability sequence determined from reversal potentials was F- greater than I- greater than SCN- greater than Br- greater than Cl- greater than glucuronate greater than NO3- greater than aspartate. This was different from the conductance sequence (Cl- greater than SCN- = F- greater than Br- greater than NO3- greater than I- greater than glucuronate greater than aspartate), indicating interaction of ions within the pore of the channel. Consistent with this was the observation of anomalous mole fraction dependence with a mixed solution of thiocyanate and chloride. 3. In addition to the main open level (about 400 pS; excised patch, symmetrical 165 mM-Cl-), three subconductance levels and one supraconductance level were observed. These were concluded to be integral components of the same channel based on coincidence of appearance and identical permeabilities. 4. The channel is voltage dependent, with open probability in excised patches increasing with more positive potentials. The channel was reversibly blocked in a voltage-dependent manner by SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid), a stilbene derivative, on the cytoplasmic face. 5. Several differences were noted between cell-attached and excised-patch recordings. The multiple conductance level channel was less frequently seen in cell-attached patches but could often be induced to appear by prolonged application of positive voltages. This induced channel in attached patches showed an altered voltage dependence which could be partially mimicked in excised patches by including cyclic AMP and ATP in the solution on the cytoplasmic side of the membrane.
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PMID:Anion channels with multiple conductance levels in a mouse B lymphocyte cell line. 247 28

On the fifth day following inoculation into an unstirred liquid surface culture, Penicillium atrovenetum abruptly, and reproducibly, secretes large quantities (2 g/liter) of the toxic antibiotic 3-nitropropionate into the medium. Concomitantly and with the same time course, crude extracts of the fungus acquire the ability to catalyze the oxidation of 3-nitropropionate by O2. We purified this activity some 300-fold to homogeneity and find it to be a soluble, dimeric (Mr = 73,000) flavoprotein oxidase having FMN as prosthetic group with lambda max = 363 and 433 nm. The preferred substrates are propionate-3-nitronate (3-NP-2) and O2 while the reaction products are malonate semialdehyde, NO2-, NO3-, O2-., and H2O2. Of 13 nitronates tested only butyrate-4-nitronate is more than 2% as reactive as 3-NP-2. 3-NP-2 (0.1 mM) rapidly reduces E-FMN anaerobically to E-FMNH., the flavin semiquinone (t1/2 less than 5 s), but reduces E-FMNH. to the fully reduced enzyme (E-FMNH2) very slowly (t1/2 approximately 900 s). The steady state turnover number with 0.1 mM 3-NP-2 and infinite O2 is 350 s-1. Therefore, the enzyme must oscillate almost exclusively between E-FMN and E-FMNH. during aerobic turnover. (Formula: see text). The complicated and non-integral reaction stoichiometry provides further support for this free radical mechanism. Each mole of 3-NP-. generated enzymatically initiates the nonenzymatic autoxidation of at least 2.2 mol of 3-NP-2 through a free radical chain reaction. An appropriate name for the newly characterized enzyme is propionate-3-nitronate oxidase.
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PMID:Propionate-3-nitronate oxidase from Penicillium atrovenetum is a flavoprotein which initiates the autoxidation of its substrate by O2. 366 82

Single-channel currents through GABA- and glycine-activated chloride channels of post-natal tissue-cultured hippocampal neurons were measured to determine their anion selectivity and their concentration dependence of permeation. Current-voltage relations for both agonists displayed rectification with single-channel conductance increasing at positive potentials. Permeabilities determined from reversal potentials were maximal for anions with a diameter of about 4 A. Larger diameter anions had lower permeabilities, consistent with an approximate pore diameter of 6 A for both agonist-activated channels. The permeability for anions of similar size was greatest for those ions with a more symmetrical charge distribution (e.g. NO3- > Bicarbonate-). The permeability sequence was SCN- > NO3- > I- > Br- > Cl- > Formate- > Acetate- > Bicarbonate- > Gluconate- > F- > Phosphate-, whereas the conductance sequence for anion efflux was Cl- > Br- > NO3- > I- > SCN- > Formate- > Acetate- > Bicarbonate- > Gluconate- > F- > Phosphate-. These results suggest that the ions interact with sites within the channel, with hydration forces contributing an important component to the barrier for ion entry into the channel. The spherically symmetrical halides displayed an exponential relation between relative permeability and hydration energy. Concentration dependence of conductance for Cl- channels in symmetrical Cl- solutions with agonist in the pipette showed an increase at positive potentials and a decrease at negative potentials. GABA- and glycine-activated channels also exhibited anomalous mole-fraction effects in a mixture of Cl- and SCN-. These results suggest that both agonist-activated channels act as multi-ion pathways and have similar permeation characteristics.
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PMID:Anion permeation in GABA- and glycine-gated channels of mammalian cultured hippocampal neurons. 769 Apr 84

Chloride channels of the ClC family are important for the control of membrane excitability, cell volume regulation, and possibly transepithelial transport. Although lacking the typical voltage-sensor found in cation channels, gating of ClC channels is clearly voltage-dependent. For the prototype Torpedo channel ClC-0 (refs 11-15) we now show that channel opening is strongly facilitated by external chloride. Other less permeable anions can substitute for chloride with less efficiency. ClC-0 conductance shows an anomalous mole fraction behaviour with Cl-/NO3- mixtures, suggesting a multi-ion pore. Gating shows a similar anomalous behaviour, tightly linking permeation to gating. Eliminating a positive charge at the cytoplasmic end of domain D12 changes kinetics, concentration dependence and halide selectivity of gating, and alters pore properties such as ion selectivity, single-channel conductance and rectification. Taken together, our results strongly suggest that in these channels voltage-dependent gating is conferred by the permeating ion itself, acting as the gating charge.
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PMID:Gating of the voltage-dependent chloride channel CIC-0 by the permeant anion. 784 66

The abilities of platinum(IV) complexes to induce the biosynthesis of metallothionein (MT) were investigated in rabbits given injections s.c. of sodium chloroplatinate (Na2PtCl6) and iproplatin (cis-dichloro-bis-isopropylamine-trans-dihydroxylplatinum IV). It is revealed for the first time that both complexes can induce MT synthesis in the liver and the kidney, but the induction ability was weaker compared to Zn2+ compounds. The induced MT was purified and identified. The hepatic MT resulting from Na2PtCl6 injection only contained Zn, whereas the hepatic MT from iproplatin injection and the renal MT from injection of both complexes contained 4-5 Zn and 1-2 Pt per mole of protein, and the renal MT also contained 1-2 Cu per mole of protein. The oxidation state of platinum in the MT is +2 as determined by X-ray photoelectron spectroscopic measurements. Pretreatment with Zn(NO3)2 elevated the levels of MT, but the binding of Pt to MT was significantly less compared to that without Zn(NO3)2 pretreatment. The data obtained from the amino acid composition analysis were consistent with the theoretical values. Upon these bases, the role of MT in relation to its involvement in the metabolism of Pt(IV) complexes and the mechanism of drug resistance to the Pt(IV) complexes as antitumor agents are discussed.
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PMID:Interaction of sodium chloroplatinate and iproplatin with metallothionein in vivo. 913 Mar 91

A distinctive feature of the voltage-dependent chloride channels ClC-0 (the Torpedo electroplaque chloride channel) and ClC-1 (the major skeletal muscle chloride channel) is that chloride acts as a ligand to its own channel, regulating channel opening and so controlling the permeation of its own species. We have now studied the permeation of a number of foreign anions through ClC-1 using voltage-clamp techniques on Xenopus oocytes and Sf9 cells expressing human (hClC-1) or rat (rClC-1) isoforms, respectively. From their effect on channel gating, the anions presented in this paper can be divided into three groups: impermeant or poorly permeant anions that can not replace Cl- as a channel opener and do not block the channel appreciably (glutamate, gluconate, HCO3-, BrO3-); impermeant anions that can open the channel and show significant block (methanesulfonate, cyclamate); and permeant anions that replace Cl- at the regulatory binding site but impair Cl- passage through the channel pore (Br-, NO3-, ClO3-, I-, ClO4-, SCN-). The permeability sequence for rClC-1, SCN- approximately ClO4- > Cl- > Br- > NO3- approximately ClO3- > I- >> BrO3- > HCO3- >> methanesulfonate approximately cyclamate approximately glutamate, was different from the sequence determined for blocking potency and ability to shift the Popen curve, SCN- approximately ClO4- > I- > NO3- approximately ClO3- approximately methanesulfonate > Br- > cyclamate > BrO3- > HCO3- > glutamate, implying that the regulatory binding site that opens the channel is different from the selectivity center and situated closer to the external side. Channel block by foreign anions is voltage dependent and can be entirely accounted for by reduction in single channel conductance. Minimum pore diameter was estimated to be approximately 4.5 A. Anomalous mole-fraction effects found for permeability ratios and conductance in mixtures of Cl- and SCN- or ClO4- suggest a multi-ion pore. Hydrophobic interactions with the wall of the channel pore may explain discrepancies between the measured permeabilities of some anions and their size.
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PMID:Permeation and block of the skeletal muscle chloride channel, ClC-1, by foreign anions. 956 3


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