首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
A calcium sensitive univalent cation channel could be formed by lysotriphosphoinositide on an artificial bilayer membrane made of oxidized cholesterol. The modified membrane was selectively permeable to univalent cations, but was only very sparingly permeable to anions or divalent cations. Selectivity sequence among group IA cations was Rb+ greater than Cs+ greater than Na+ greater than K+ greater than Li+. The conductance of the membrane was increased up to a value of about 10-2 ohm-1/cm2 with an increase in the concentration of univalent cation, and was drastically depressed by a relatively small increase in the concentration of calcium ion or other divalent cations. The sequence of depressing efficiency among divalent cations was Zn+ greater than Cd2+ greater than Ca2+ greater than Sr2+ greater than Mg2+.  相似文献   

2.
Summary A nonselective cation channel activated by patch excision was characterized in inside-out patches from spiny lobster olfactory receptor neurons. The channel, which was permeable to Na+, K+ and Cs+, had a conductance of 320 pS and was weakly voltage dependent in the presence of micromolar divalent cations. Millimolar internal divalent cations caused a voltage-and concentration-dependent block of Na+ permeation. Analysis of the voltage dependence indicated that the proportion of the membrane's electric field sensed by Mg2+ was >1, suggesting that the channel contains a multi-ion pore. Internal divalent cations also reduced the frequency of channel opening in a concentration-dependent, but not voltage-dependent, manner, indicating that different cation binding sites affect gating and conductance. While block of gating prevented determining if internal divalent cations permeate the channel, a channel highly permeable to external divalent cations was observed upon patch excision to the inside-out configuration. The monovalent and divalent cation conductances shared activation by patch excision, weak voltage dependence, and steady-state activity, suggesting that they are the same channel. These data extend our understanding of this type of channel by demonstrating permeation by monovalent cations, detailing Mg2+ block of Na permeation, and demonstrating the channel's presence in arthropods.  相似文献   

3.
The effects of monovalent cations - inorganic alakali metal cations and organic quanternary ammonium cations - and monovalent inorganic anions on ADP-induced aggregation of bovine platelets were investigated. In the presence of K+, Rb+, Cs+, choline or tetramethylammonium, aggeregation proceeded. However, aggregation was markedly restricted in media containing Li+, Na+, tetrabutylammonium or dimethyldibenzylammonium. With anions, aggregation proceeded in the order Cl > Br > I > Clo4 > SCN. The effects of cations significantly depended on Ca2+ concentration, whereas those of the anions depended little of Ca2+. Anions such as SCN and ClO4 markedly decreased the fluorescence of the surface charge probe 2-p-tuluidinylnaphthalene-6-sulfonate, whereas cations had less pronouced effects. The relative effects of the anions on the fluorescence were consistent with their relative inhibitory effects on aggregation. These results suggest that inhibition of platelet aggregation by the anions is due to a change in the surface change of the platelet plasma membrane. On the other hand, kinetic analysis suggests that the effects of monovalent cations on platelet aggregation are due to their competition with Ca2+ during the process of aggregation.  相似文献   

4.
The cation exchange properties of cell walls isolated from collard (Bassica oleracea var acephala D.C.) leaves were investigated. Cation sorption on cell walls was described by mass-action expressions of ion exchange, rather than by the traditional Donnan equilibrium. The mass-action expressions enable the selectivity of the wall for one cation over another to be determined unambiguously from ion exchange isotherms. We found that: (a) the cation composition of the wall varied as a function of the solution cation concentration, solution cation composition, and pH in a way predicted by mass action; (b) the affinity of the wall for divalent cations increased as the equivalent fraction of divalent cation on the wall increased, and as the concentration of divalent cations in solution increased; (c) the selectivity of the wall for any metal cation pair was not altered by the concentration of H+ in solution or on the wall; (d) H+ sorption on the wall may be treated as a cation exchange reaction making it possible to calculate the relative affinity of the wall for metal cation pairs from H+-metal (Me) titration curves; and (e) the relative affinity of the wall for the cations we studied was: H+ (K+ ≥ Ca2+) > Mg2+. A cation-exchange model including surface complexes is consistent with observed cation selectivity. We conclude that metal cations interact with the wall to minimize or eliminate long-range electrostatic interactions and suggest that this may be due to the formation of site-specific cation-wall surface complexes.  相似文献   

5.
Summary Leakage of ions (Na+, K+) and phosphorylated metabolites (phosphorylcholine, 2-deoxyglucose 6-phosphate) through membrane lesions in intact cells or in cells modified by pore-forming agent has been studied. Leakage from intact cells isinduced by protons and by divalent cations such as Cu2+, Cd2+ or Zn2+. Leakage from agent-modified cells—or across phospholipid bilayers modified by agent—isprevented by low concentrations of the same cations and by higher concentrations of Ca2+, Mn2+ or Ba2+; Mg2+, dimethonium, spermine, or spermidine are virtually ineffective. The relative efficacy of a particular cation (e.g. Ca2+) depends more on cell type than on the nature of the pore-forming agent. The predominant effect is on binding of cation to specific sites, not on surface charge. Surface charge, on the other hand, does affect leakage from agent-modified cells in that suspension in nonionic media reduces leakage, which can be restored by increasing the ionic strength: univalent (Na+, K+, Rb+, NH 4 + ) and divalent (Mg2+, dimethonium) cations are equally effective; addition of protons or divalent cations such as Zn2+ to this system inhibits leakage. From this and other evidence here presented it is concluded that leakage across membranes is modulated by the presence of endogenous anionic components: when these are in the ionized state, leakage is favored; when unionized (as a result of protonation) or chelated (by binding to divalent cation), leakage is prevented. It is suggested that such groups are exposed at the extracellular face of the plasma membrane.  相似文献   

6.
The specific modulation by three cations, Ca2+, Mg2+, and tetracaine of the equilibrium exchange of SO42− across the erythrocyte membrane was investigated. While external calcium had no effect on SO42− exchange, internal calcium, and external calcium in the presence of 10 μM A23187 were found to be potent inhibitors of the exchange reaction. The apparent inhibition constants (K1) for Ca2+ were calculated to be 6.1 μM and 5 μM for the above two conditions, respectively.Unlike Ca2+, Mg2+ was shown to be a weak activator of SO42− exchange with an apparent dissociation constant of 3.6 μM. Competition experiments demonstrated that the Ca2+ and Mg2+ sites associated with anion transport are distinct and noninteracting.Tetracaine, a cation at neutral pH, was also found to be an inhibitor of SO42− exchange with an apparent K1 of 0.8 mM. Although tetracaine was observed to displace calcium from non-specific sites on the erythrocyte membrane, it showed no effect on the apparent inhibition constant of Ca2+ for SO42− exchange. Thus, the Ca2+ and tetracaine sites also appear to be independent. The difficulty of situating three mutually independent sites on a single subunit protein, i.e., band 3, is considered.Using the experimental data obtained from five individuals, the concentration of free calcium in the red cell cytoplasm was calculated to range from 0.2 to 0.7 μM. This concentration was sufficient to reduce SO42− exchange only 3–8%. It was concluded that calcium inhibition of anion exchange, and, hence, impairment of CO2 transport, may be physiologically significant only in senescent cells and in certain types of anemia where calcium concentrations are significantly increased.  相似文献   

7.
The (K+,Mg2+)-ATPase was partially purified from a plasma membrane fraction from corn roots (WF9 × Mol7) and stored in liquid N2 without loss of activity. Specific activity was increased 4-fold over that of the plasma membrane fraction. ATPase activity resembled that of the plasma membrane fraction with certain alterations in cation sensitivity. The enzyme required a divalent cation for activity (Co2+ > Mg2+ > Mn2+ > Zn2+ > Ca2+) when assayed at 3 millimolar ATP and 3 millimolar divalent cation at pH 6.3. When assayed in the presence of 3 millimolar Mg2+, the enzyme was further activated by monovalent cations (K+, NH4+, Rb+ Na+, Cs+, Li+). The pH optima were 6.5 and 6.3 in the absence and presence of 50 millimolar KCl, respectively. The enzyme showed simple Michaelis-Menten kinetics for the substrate ATP-Mg, with a Km of 1.3 millimolar in the absence and 0.7 millimolar in the presence of 50 millimolar KCl. Stimulation by K+ approached simple Michaelis-Menten kinetics, with a Km of approximately 4 millimolar KCl. ATPase activity was inhibited by sodium orthovanadate. Half-maximal inhibition was at 150 and 35 micromolar in the absence and presence of 50 millimolar KCl. The enzyme required the substrate ATP. The rate of hydrolysis of other substrates, except UDP, IDP, and GDP, was less than 20% of ATP hydrolysis. Nucleoside diphosphatase activity was less than 30% of ATPase activity, was not inhibited by vanadate, was not stimulated by K+, and preferred Mn2+ to Mg2+. The results demonstrate that the (K+,Mg2+)-ATPase can be clearly distinguished from nonspecific phosphohydrolase and nucleoside diphosphatase activities of plasma membrane fractions prepared from corn roots.  相似文献   

8.
The internal cation levels of chloroplasts isolated from a green sea alga, Bryopsis maxima, were studied. Atomic absorption spectroscopy, combined with the determination of the sorbitol-impermeable and water-permeable spaces, revealed that chloroplasts contain an extremely high concentration of K+ and high levels of Na+, Mg2+ and Ca2+. A method was developed to estimate the thermodynamic activities of monovalent and divalent cations present in chloroplasts. pH changes induced by the addition of an ionophore (plus an H+ carrier), which makes the outer limiting membranes of chloroplasts permeable to both a cation and H+, were determined. Provided that the external pH was set equal to the internal pH, the internal concentration of the cation was estimated by determining the external cation concentration which gave rise to no electrochemical potential difference of the cation and hence no pH change on addition of the ionophore. The internal pH was determined by measuring distributions of radioactive methylamine and 5,5-dimethyloxazolidine-2,4-dione between the chloroplast and medium (Heldt, H.W., Werdan, K., Milovancev, M. and Geller, G. (1973) Biochim. Biophys. Acta 314, 224–241). The internal pH was also estimated by measuring pH changes caused by the disruption of the outer limiting membrane with Triton X-100. The results indicate that a significant part of the monovalent cations and most of the divalent cations are attracted into a diffuse layer adjacent to the negatively charged surfaces of membranes and proteins, or form complexes with organic and inorganic compounds present in the intact chloroplasts.  相似文献   

9.
Adenosine triphosphatase from soybean callus and root cells   总被引:9,自引:9,他引:0       下载免费PDF全文
The ATPase activity of a membrane fraction from soybean (Glycine max L.) root and callus cells, presumed to be enriched in plasma membrane, has been characterized with respect to ion stimulation, pH requirement, and nucleotide specificity. The enzyme from both sources was activated by divalent cations (Mg2+ > Mn2+ > Zn2+ > Ca2+ > Sr2+) and further stimulated by monovalent salts. Preparations from root cells were stimulated by monovalent ions according to the sequence: K+ > Rb+ > Choline+ > Na+ > Li+ > NH4+ > Cs+ > tris+. Membrane preparations from callus cells showed similar stimulatory patterns except for a slight preference for Na+ over K+. No synergism between K+ and Na+ was found with preparations from either cell source.  相似文献   

10.
Summary The surface charge density resulting from the adsorption of hydrophobic anions of dipicrylamine onto dioleyl-lecithin bilayer membranes has been measured directly using a high field pulse method. The surface charge density increases linearly with adsorbate concentration in the water until electrostatic repulsion of impinging hydrophobic ions by those already adsorbed becomes appreciable. Then Gouy-Chapman theory predicts that surface charge density will increase sublinearly, with the power [z +/(z ++2)] of the adsorbate concentration, wherez + is the cation valence of the indifferent electrolyte screening the negatively charged membrane surface. The predicted 1/3 and 1/2 power laws for univalent and divalent cations, respectively, have been observed in these experiments using Na+, Mg++, and Ba++ ions. Gouy-Chapman theory predicts further that the change from linear to sublinear dependence takes place at a surface charge density governed by the static dielectric constant of water and the concentration of indifferent electrolyte. Quantitative agreement with experiment is obtained at electrolyte concentrations of 10–4 m and 10–3 m, but can be maintained at higher concentrations only if the aqueous dielectric constant is decreased. A transition field model is proposed in which the Gouy-Chapman theory is modified to take account of dielectric saturation of water in the intense electric fields adjacent to charged membrane surfaces.  相似文献   

11.
Summary Outward rectifying. cation channels were observed in the epithelial cells of the urinary bladder of the toad.Bufo marinus. As studied in isolated cells using the patch-clamp technique, the channel has an average conductance of 24 and 157 pS for pipette potentials between 0 and +60 mV and –60 to –100 mV, respectively, when the major cation in both bath and pipette solutions is K+. The conductance of the cannel decreasen with increasing dehydration energy of the permeant monovalent cation in the oder Rb+=K+>Na+>Li+. Reversal potentials near zero under biionic conditions imply that the permeabilities for all four of these cations are smiliar. The channel is sensitive to quinidine sulfate but not to amiloride. It shares several pharmacological and biophysical properties with an outwardly-rectifying, vasopressin-sensitive pical K+ conductive pathway described previously for the toad urinary bladder. We demonstrate, in both single-channel and whole-bladder studies, that the outward rectification is a consequence of interaction of the chanel with extracellular divalent cations, particularly Ca2+, which blocks inward but not outward current. Various divalent cations impart different degrees of outward rectification to the conductive pathway. Concentrations of Mg2+ and Ca2+ required for halfmaximal effect are 3×10–4 and 10–4 m, resopectively. For Co2+ the values are 10–6 m at +50 mV and a 10–4 m at +200 mV. The mechanism of blockade by divalent cations is not established, but does not seem to involve a voltage-dependent interaction in which the blocker penetrates the transmembrane electric field. In the absence of divalent cations in the mucosal solution, the magnitudes of inward current carried by Rb+, K+, Na+ and Li+ through the apical K+ pathway at any transepithelial voltage, are in the same order as in the single-channel studies. We propose that the cation channel observed by us in isolated epithelial cells is the single-channel correlate of the vasopressin-sensitive apical K+ conductive pathway in the toad urinary bladder and is also related to the oxytocin- and divalent cation-sensitive apical condictivity observed in frog skin and urinary bladder.  相似文献   

12.
The pH within isolated Triton WR 1339-filled rat liver lysosomes was determined by measuring the distribution of [14C]methylamine between the intra- and extralysosomal space. The intralysosomal pH was found to be approximately one pH unit lower than that of the surrounding medium. Increasing the extralysosomal cation concentration lowered the pH gradient by a cation exchange indicating the presence of a Donnan equilibrium. The lysosomal membrane was found to be significantly more permeable to protons than to other cations. The relative mobility of cations through the lysosomal membrane is H+ ? Cs+ > Rb+ > K+ Na+ > Li+ ? Mg2+, Ca2+. The presented data suggest that the acidity within isolated Triton WR 1339-filled lysosomes is maintained by: (1) a Donnan equilibrium resulting from the intralysosomal accumulation of nondifussible anions and (2) a selective permeability of the lysosomal membrane to cations.  相似文献   

13.
This paper examines the applicability of the Nernst-Planck approach in treating the relationship between the initial rates at which critical ions interexchange across permselective membranes in bi-ionic systems and the rates of self-exchange of these ions across the same membranes. Data are presented for five species of univalent cations with two types of cation permeable membranes, a polystyrene sulfonic acid-collodion matrix membrane, and an oxidized collodion membrane; five species of univalent anions were studied with a protamine-collodion matrix anion permeable membrane. Except with systems involving H+ ion, the experimentally found relationships between the rates of interexchange and the rates of self-exchange were in agreement, in most cases within ±5%, with the values calculated from an expression in which interaction between the critical ions in the membrane is not taken into account. In systems with H+ ion, the experimental rates of interexchange were from 27% to 40% less than calculated values.  相似文献   

14.
Mg2+, Ca2+ and Mn2+ were found to act as activators of the ATP-dependent surface reaction, leading to head-to-head association in bull spermatozoa. Ca2+ was more efficient than Mg2+, while Zn2+, like Na+ + K+ in combination with Mg2+, seemed to have no such effect. High ionic strength induced head-to-head association, as did higher concentrations of Mg2+ and Ca2+ than those necessary for the activation of ATP, Ca2+ acting in a lower conc. than Mg2+. To this effect was added that of the ATP-dependent reaction when ATP was also present. As activators, Mg2+ and Ca2+ did not potentiate each other; their effects were cumulative when the ions acted together.When the ATP concentration within the range 1 × 10−5 to 8 × 10−5 M was increased stepwise in the presence of 2 × 10−5 M Mg2+ or Ca2+, the association resulting from each single concentration step progressively increased. At low cation concentrations, the increase was about the same for the two cations: at higher concentrations it was much steeper in the presence of Ca2+ than in that of Mg2+. In the latter case, it was not statistically significant above 4 × 10−5 M ATP.Increasing the cation concentration in the range 1 × 10−5 to 4 × 10−5 M in the presence of 2 × 10−5 M ATP produced an immediate high increase in association, which was followed by a lower increase. The optimum concentration ratio for Mg2+:ATP was at least 1:1 and for Ca2+: ATP at least 1.5:1.Oubain, containing enone structure, abolishes association.  相似文献   

15.
Ionophore A23187-mediated Ca2+-induced oscillations in the conductance of the Ca2+-sensitive K+ channels of human red cells were monitored with ion specific electrodes. The membrane potential was continuously reflected in CCCP-mediated pH changes in the buffer-free medium, changes in extracellular K+ activity were followed with a K+-selective electrode, and changes in the intracellular concentration of ionized calcium were calculated on the basis of cellular 45Ca content. An increased cellular 45Ca content at the successive minima of the oscillations where the K+ channels are closed indicates that the activation of the channels might be a (dCa2+/dt)-sensitive process and that accommodation to enhanced levels of intracellular free calcium may occur. An incipient inactivation of the K+ channels at intracellular ionized calcium levels of about 10 μM and a concurrent membrane potential of about −65 mV was observed. At a membrane potential of about −70 mV and an intracellular concentration of about 2·10−4M no inactivation of K+ channels took place. Inactivation of the K+ channels is suggested to be a compound function of the intracellular level of free calcium and the membrane potential. The observed sharp peak values in cellular 45Ca content support the notion that a necessary component of the oscillatory system is a Ca2+ pump operating with a significant delay in the activation/inactivation process in response to changes in cellular concentration of ionized calcium.  相似文献   

16.
The rhizotoxicities of Al3+ and of La3+ to wheat (Triticum aestivum L.) were similarly ameliorated by cations in the following order of effectiveness: H+ ≈ C3+ > C2+ > C1+. Among tested cations of a given charge, ameliorative effectiveness was similar except that Ca2+ was slightly more effective than other divalent cations and H+ was much more effective than other monovalent cations. H+ rhizotoxicity was also ameliorated by cations in the order C3+ > C2+ > C1+. These results suggest a role for cell-surface electrical potential in the rhizotoxicity of Al3+, La3+, H+, and other toxic cations: negatively charged cell surfaces of the root accumulate the toxic cations, and amelioration is effected by treatments that reduce the negativity of the cell-surface electrical potential by charge screening or cation binding. Membrane-surface activities of free Al3+ or La3+ computed according to a Gouy-Chapman-Stern model correlated well with growth inhibition, which correlated only poorly with Al3+ or La3+ activities in the external medium. The similar responses of Al-intoxicated and La-intoxicated roots to ameliorative treatments provide evidence that Al3+, rather than AlOH2+ or Al(OH)2+, is the principal toxic species of mononuclear Al. Comparisons of the responses of Al-sensitive and Al-tolerant wheats to Al3+ and to La3+ did not support the hypothesis that varietal sensitivity to Al3+ is based upon differences in cell-surface electrical potential.  相似文献   

17.
The resting membrane of a barnacle muscle fiber is mostly permeable to cations in a solution of pH 7.7 whereas it becomes primarily permeable to anions if the pH is below 4.0. Mechanisms of ion permeation for various monovalent cations and anions were investigated at pH 7.7 and 3.9, respectively. Permeability ratios were obtained from the relationship between the membrane potential and the concentration of the test ions, and ionic conductances from current-voltage relations of the membrane. The permeability sequence for anions (SCN > I > NO3 > Br > ClO3 > Cl > BrO3 > IO3) was different from the conductance sequence for anions (Br, Cl > ClO3, NO3 > SCN). In contrast, the permeability and conductance sequences were identical for cations (K > Rb > Cs > Na > Li). The results suggest that anion permeation is governed by membrane charges while cation permeation is via some electrically neutral mechanism.  相似文献   

18.
Laser Raman spectroscopy was applied to characterize structural behavior of dipalmitoyl phosphatidylcholine multibilayer systems in the presence of several cations (K+, Na+, Cs+, Rb+, Ca2+, Mg2+, Cd2+, Ba2+) and anions (Cl, Br, I, NO3, SO32−, SO42−, S2O32−, S2O82−). To evaluate the Raman-spectroscopical data quantitatively, characteristic intensity ratios, lateral and trans order parameters were used and compared. It was shown that the different trans order parameters are rather sensitive to ion-polar head group interactions and thus, they cannot give unequivocal information on the trans-gauche isomerization of hydrocarbon chains of phospholipids. The observed effects of ions on Raman spectra of phospholipid multilayers could not be explained simply on the basis of electrostatic interactions. The possible involvement of other factors (changes in polarizability, hydrogen bonds, etc.) is also discussed. It was demonstrated that the order parameters defined in different ways may result in different effectiveness sequences of ions. Of monopositive ions Na+ was found to be the most effective to influence the bilayer structure. For dipositive ions, of which Ca2+ proved to be the most effective, concentration-dependent effectiveness sequences were obtained. A plausible interpretation and some consequences of the concentration-dependent two-step binding of divalent cations were also outlined. Bilayered phospholipid structures turned out to be more responsive to anions than to most cations investigated. Interdependent actions of cations and anions, as well as the possible relevance of the charge distribution on anions are postulated.  相似文献   

19.
Summary (1) When salts are added to buffered suspensions of membrane fragments containing the fluorochrome 1-anilino-8-naphthalenesulfonate (ANS), there is an increased fluorescence. This is caused by increased binding of the fluorochrome; the intrinsic fluorescence characteristics of the bound dye remain unaltered. These properties make ANS a sensitive and versatile indicator of ion association equilibria with membranes. (2) Alkali metal and alkylammonium cations bind to membranes in a unique manner. Cs+ binds most strongly to rat brain microsomal material, with the other alkali metals in the order Cs+>Rb+>K+>Na+>Li+. The reaction is endothermic and entropy driven. Monovalent cations are displaced by other monovalent cations. Divalent cations and some drugs (e. g., cocaine) displace monovalent cations more strongly. (3) Divalent cations bind to membranes (and to lecithin micelles) at four distinct sites, having apparent association constants between 50 and 0.2mm –1. The characteristics of the titration suggest that only one species of binding site is present at any one time, and open the possibility that structural transitions of the unassociated coordination sites may be induced by divalent cation binding. Divalent cation binding at the weakest site (like monovalent cation binding) is endothermic and entropy driven. At the next stronger site, the reaction is exothermic. Monovalent cations affect divalent cation binding by reducing the activity coefficient: they do not appear to displace divalent cations from their binding sites.  相似文献   

20.
As described by others, an extracellular calcium-sensitive non-selective cation channel ([Ca2+]o-sensitive NSCC) of central neurons opens when extracellular calcium level decreases. An other non-selective current is activated by rising intracellular calcium ([Ca2+] i ). The [Ca2+]o-sensitive NSCC is not dependent on voltage and while it is permeable by monovalent cations, it is blocked by divalent cations. We tested the hypothesis that activation of this channel can promote seizures and spreading depression (SD). We used a computer model of a neuron surrounded by interstitial space and enveloped in a glia-endothelial “buffer” system. Na+, K+, Ca2+ and Cl concentrations, ion fluxes and osmotically driven volume changes were computed. Conventional ion channels and the NSCC were incorporated in the neuron membrane. Activation of NSCC conductance caused the appearance of paroxysmal afterdischarges (ADs) at parameter settings that did not produce AD in the absence of NSCC. The duration of the AD depended on the amplitude of the NSCC. Similarly, NSCC also enabled the generation of SD. We conclude that NSCC can contribute to the generation of epileptiform events and to spreading depression.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号