A cotransport system for Na+, K+ and Cl? in Ehrlich cells is described. It is insensitive towards ouabain but specifically inhibited by furosemide and other ‘high ceiling’ diuretics at concentrations which do not affect other pathways of the ions concerned. As the furosemide-sensitive fluxes of these ions are not affected by changes in membrane potential, and as their complete inhibition by furosemide does not appreciably alter the membrane potential, they appear to be electrically silent. Application of the pulse-response methods in terms of irreversible thermodynamics reveals tight coupling between the furosemide-sensitive flows of Na+, K+ and Cl? ( close to unity for all three combinations) at a stoichiometry of 1 : 1 : 2. The site for each of the ions appears to be rather specific: K+ can be replaced by Rb+ but not by other cations tested whereas Cl? can be poorly replaced by Br? but not by NO3?, in contradistinction to the Cl?-OH? exchange system. The cotransport system appears to function in cell volume regulation as it tends to make the cell swell, thus counteracting the shrinking effect of the ouabain-sensitive (Na+, K+) pump.The experiments presented could not clarify whether the cotransport process is a primary or secondary active one; while incongruence between transport and conjugated driving force seems to indicate primary active transport, it is very unlikely that hydrolysis of ATP supplies energy for the transport process, since there is no stimulation of ATP turnover observable under operation of the cotransport system. 相似文献
The balance of K+, Na+, and Cl− fluxes across the cell membrane with the Na+/K+ pump, ion channels, and Na+K+2Cl− (NKCC) and Na+-Cl− (NC) cotransport was calculated to determine the mechanism of cell shrinkage in apoptosis. It is shown that all unidirectional
K+, Na+, and Cl− fluxes; the ion channel permeability; and the membrane potential can be found using the principle of the flux balance if
the following experimental data are known: K+, Na+, and Cl− concentrations in cell water; total Cl− flux; total K+ influx; and the ouabain-inhibited pump component of the Rb+(K+) influx. The change in different ionic pathways during apoptosis was estimated by calculations based on the data reported
in the preceded paper (Yurinskaya et al., 2010). It is found that cell shrinkage and the shift in ion balance in U937 cells
induced to apoptosis with 1 μM staurosporine occur due to the coupling of reduced pump activity with a decrease in the integral
permeability of Na+ channels, whereas K+ and Cl− channel permeability remains almost unchanged. Calculations show that only a small part of the total fluxes of K+, Na+, and Cl− account for the fluxes mediated by NKCC and NC cotransporters. Despite the importance of cotransport fluxes for maintaining
the nonequilibrium steady-state distribution of Cl−, they cannot play a significant role in apoptotic cell shrinkage because of their minority and cannot be revealed by inhibitors. 相似文献
Effects of Na+ and K+ on Ca2+ transport by sarcoplasmic reticulum vesicles were studied in a medium containing high Mg2+ and ATP (2mM) and low Ca2+ (0.44μM) concentrations. Under these conditions, Na+ and K+ inhibit Ca2+ uptake. ATPase activity and membrane phosphorylation by ATP. Since the concentrations of ATP and Ca2+ used are consistent with relaxation in vivo, the results suggest that under physiological resting conditions the Ca2+ pump of the sarcoplasmic reticulum operates below its maximal capacity. 相似文献
Mechanisms underlying the tissue-specific impact of cardiotonic steroids (CTS) on cell survival and death remain poorly understood.
This study examines the role of Na+,K+-ATPase α subunits in death of Madin-Darby canine kidney (MDCK) cells evoked by 24-h exposure to ouabain. MDCK cells expressing
a variant of the α1 isoform, CTS-sensitive α1S, were stably transfected with a cDNA encoding CTS-resistant α1R-Na+,K+-ATPase, whose expression was confirmed by RT–PCR. In mock-transfected and α1R-cells, maximal inhibition of 86Rb influx was observed at 10 and 1000 μM ouabain, respectively, thus confirming high abundance of α1R-Na+,K+-ATPase in these cells. Six-hour treatment of α1R-cells with 1000 μM ouabain led to the same elevation of the [Na+]i/[K+]i ratio that was detected in mock-transfected cells treated with 3 μM ouabain. However, in contrast to the massive death of
mock-transfected cells exposed to 3 μM ouabain, α1R-cells survived after 24-h incubation with 1000 μM ouabain. Inversion of
the [Na+]i/[K+]i ratio evoked by Na+,K+-ATPase inhibition in K+-free medium did not affect survival of α1R-cells but increased their sensitivity to ouabain. Our results show that the α1R
subunit rescues MDCK cells from the cytotoxic action of CTS independently of inhibition of Na+,K+-ATPase-mediated Na+ and K+ fluxes and inversion of the [Na+]i/[K+]i ratio. 相似文献
Na+, K+ and Cl? concentrations () and activities (), and mucosal membrane potentials () were measured in epithelial cells of isolated bullfrog (Rana catesbeiana) small intestine. Segments of intestine were stripped of their external muscle layers, and bathed (at 25°C and pH 7.2) in oxygenated Ringer solutions containing 105 mM Na+ and Cl? and 5.4 mM K+. Na+ and K+ concentrations were determined by atomic absorption spectrometry and Cl? concentrations by conductometric titration following extraction of the dried tissue with 0.1 M HNO3. 14C-labelled inulin was used to determine extracellular volume. was measured with conventional open tip microelectrodes, with solid-state Cl?-selective silver microelectrodes and and with Na+- and K+-selective liquid ion-exchanger microelectrodes. The average recorded was ?34 mV. , and were 51, 105 and 52 mM. The corresponding values for , and were 18, 80 and 33 mM. These results suggest that a large fraction of the cytoplasmic Na+ is ‘bound’ or sequestered in an osmotically inactive form, that all, or virtually all the cytoplasmic K+ behaves as if in free solution, and that there is probably some binding of cytoplasmic Cl?. significantly exceeds the level corresponding to electrochemical equilibrium across the mucosal and baso-lateral cell membranes. Earlier studies showed that coupled mucosal entry of Na+ and Cl? is implicated in intracellular Cl? accumulation in this tissue. This study permitted estimation of the steady-state transapical Na+ and Cl? electrochemical potential differences (Δμ̄Na and Δμ̄Cl). Δμ̄Na (?7000 J · mol?1; cell minus mucosal medium) was energetically more than sufficient to account for Δμ̄Cl (1000–2000 J · mol?1). 相似文献
The effect of changing [K+], [Na+] and [Cl?] in nutrient solution was studied in bullfrog antrum with and without HCO3? in nutrient. In 25 mM HCO3? (95% O2/5% CO2) and in zero HCO3? (100% O2), nutrient pH was maintained at 7.3. Changing from 4 to 40 mM K+ or from 81 to 8.1 mM Cl? gave a decrease 10 min later in transmucosal PD (nutrient became more negative) — a normal response. These responses were less in zero than in 25 mM HCO3?. A decrease from 102 to 8 mM Na+ decreased PD (anomalous response of electrogenic NaCl symport). This effect was attenuated or eliminated in zero HCO3?. In contrast, change from 4 to 40 mM K+ gave initial anomalous PD response and change from 102 to 8 mM Na+, initial normal PD response with either zero or 25 mM HCO3?. Both responses were associated with (Na+ + K+)-ATPase pump and were greater in zero than in 25 mM HCO3?. Initial PD increases in zero HCO3? are explained as due to increase in the resistance of passive conductance and/or NaCl symport pathways. Thus, removal of HCO3? modifies conductance pathways of nutrient membrane. 相似文献
Vanadium(V)-induced hydrolyses of triphosphates in aqueous solutions were initiated in two ways: (1) oxidizing vanadium(IV)-polyphosphate complexes to produce metastable vanadium(V) complexes; (2) forming VO2+-polyphosphate complexes by acidification of solutions of VO43? and polyphosphate to yield equilibrium mixtures of V(V), polyphosphate, and their complexes. Hydrolysis rates for the complexes formed at 40°C ? T ? 25°C follow the order V2PPPi = 2(VPPPi) ≌ (VO2) ATP ? V(PPPi)2 ≌ PPPi. The hydrolysis of (V(V))2(PPPi) was not very temperature sensitive; the activation enthalpy appears small and the activation entropy large and negative. Mechanistic studies reveal that requirements for the activated state in metal-ion-catalyzed hydrolysis of polyphosphates include monodentate polyphosphate ligated cis to H2O or OH? in the coordination sphere of the metal ion: 相似文献
Summary Electron microscopy study shows that cytochalasin treatment of the mullet damages the microfilaments system in the apex of gill ionocytes: the microfilaments are reduced in number and shortened. Cytochalasin causes a reduction of transgill potential difference and an increase of the Na+ and Cl– blood concentration, of the diffusional water permeability of the gill, of the Na+ branchial influx and of Cl– efflux. The increase of the Na+ influx may result in a reduction of the Na+ net excretion flux compared to the control. The increased permeability in cytochalasin treated fish facilitates the Cl– entry probably leading to a reduction of the net Cl– excretion. The partial inhibition of the K+ dependent components of Na+ and Cl– effluxes also contributes to the reduction of Na+ and Cl– excretion. The role of microfilaments in the mechanisms of ionic excretion by the gill is discussed. 相似文献
The improved methods for the preparation of valency hybrid hemoglobins, (α3+β2+)2 and (α2+β3+)2 were presented. The (α3+β2+)2 valency hybrid was separated from the solutions of partially reduced methemoglobin with ascorbic acid, by using CM 32 column chromatography. The (α2+β3+)2 valency hybrid was also isolated from hemoglobin solutions, which were partially oxidized with ferricyanide, by chromatography on CM 32 column. These valency hybrid hemoglobins were found to be single on isoelectric focusing electrophoresis. Present procedures are very simple and are suitable for the bulk preparation of (α3+β2+)2 and (α2+β3+)2 valency hybrids. 相似文献