Predicting Cation Ratios in Corn from Saline Solution Composition

The response of Zea mays (L.) cv. Pioneer 3906 to nutrient culturesthat varied in Na/Ca ratios was studied at 5 different salinitylevels; 0, 0.1, –0.2, 0.3 and –0.4 MPa. Principlesof ion exchange theory were used to relate the cation composition(Ca, Mg and Na) in the shoots to activity ratios in the rootmedia. The data are expressed using the Gapon convention wherethe selectivity coefficient is related to the equivalent fractionof the exchange ions. Plots of the cation ratios of Ca/Na, Ca/Mgand Mg/Na can be represented by a single line regardless ofsalinity level. This information supports the concept derivedfrom physiological studies that the uptake of Ca, Mg and Naby salt-stressed plants is passive. The data suggest that theratios of these cations in corn shoots can be predicted forvarious salinities from solution composition of the root mediaand shoot ion concentrations at a single salinity level. Key words: Gapon equation, ion exchange, selectivity, cation uptake     

CHARACTERISTICS OF SALTS SECRETED BY TAMARIX APHYLLA

The composition and concentration of salts secreted by the salt glands of Tamarix aphylla L. grown under controlled nutrient conditions were determined. Eight ions, Na, K, Mg, Ca, Cl, NO₃, HCO₃, and SO₄, constituted 99 % + of the dry weight of salts secreted by plants grown on half-strength Hoagland's solution. The divalent cations Mg and Ca accounted for most of the cations; HCO₃ comprised about 60 % of the anions. The micronutrients B, Mn, Cu, Zn, and Mo were present in enriched concentrations in the secretion. The composition of the secretions was highly dependent on the composition of the root environment. The predominating cation in the saline culture solutions was also the predominant cation secreted. The accompanying anion in the culture solution influences the cation composition of the secreted salt. The concentration of the salt gland secretion averaged 0.5n , a 50-fold increase in concentration over the nutrient solution in which the plants were grown.     

Use of cation/anion exchange membranes for multi-element testing of acidic soils

We have developed a technique using cation/anion exchange resins which allows the simultaneous extraction from soil of Ca, Mg, K, Mn, Al and P. Ions are extracted by shaking soil with resin in distilled water. The resin is separated from the soil and ions are desorbed from the resin using an acid/salt solution. Concentrations of ions in solution are then determined by conventional means. Concentrations of ions extracted by the resin method were compared with concentrations determined by commonly-used analytical procedures. Except for Al, concentrations of ions extracted by the resin procedures correlated well with conventional extraction and analytical procedures. The resin membrane method offers considerable speed and cost advantages over conventional methods.     

Magnesium uptake by Al-stressed maize plants with special emphasis on cation interactions at root exchange sites

As aluminium (Al) severely inhibits magnesium (Mg) uptake by many plant species, Mg uptake and Mg-Al interactions in maize (Zea mays L.) were studied in a series of short and long-term experiments. A relationship between Mg uptake and the degree of Mg saturation of exchange or binding sites of the root apoplast (root-CEC) was studied by growing plants in solutions containing: (i) different concentrations of Al, calcium (Ca) and hydrogen (H) ions; and (ii) a number of organic complexes of Al. In short-term experiments, Ca had little effect on the Mg nutrition of maize plants. However, with increasing concentrations of Al and H ions in nutrient solution, there was a decrease in both the degree of Mg saturation of root-CEC and Mg uptake. Effects of pH on cation (H, Al, Mg, Ca) binding at the root apoplasm were pronounced and complicated because of a simultaneous change of H ion concentration, effective root-CEC and Al speciation. The behaviour of Al as organic Al complexes differed from that supplied as aluminium chloride (AlCl₃). In the presence of organo-Al complexes, less Mg was replaced from apoplastic binding sites and Mg uptake was inhibited less severely than with AlCl₃. In a long-term experiment, Al-citrate, in contrast with AlCl₃, was not phytotoxic to maize, expressed by the lack of any inhibition of shoot biomass production.     

In situ ion exchange resin bags to estimate forest site quality

Cation and anion exchange resin bags were placed just under the humus layer at five adjacent forest sample sites with differing site quality classes in order to assess the available nutrient supply. For comparison, humus samples were collected from the same sites. Nutrients were extracted from humus samples by conventional extraction methods and by shaking together with ion exchange resin bags. Ca and Mg corresponded best to differences in site quality class, of all analysed ions in thein situ resin bag eluates. Thein situ resin bag adsorption of NH₄−N, Na and Mn also showed a positive correlation with site quality. The adsorption of PO₄−P was negatively correlated to site quality class. Inadequate amounts of exchange resin, or leaving resin bagsin situ for too long a time result in the replacement of already adsorbed ions by ions with higher ion exchange constants.     

Aluminium accumulation in root cell walls coincides with inhibition of root growth but not with inhibition of magnesium uptake in Norway spruce

High levels of aluminium in the soil solution of forest soils cause stress to forest trees. Within the soil profile, pH and aluminium concentration in the soil solution vary considerably with soil depth. pH strongly influences the speciation of A1 in solution, and is a factor when considering toxicity of A1 to roots. Norway spruce ( Picea abies [L.] Karst.) seedlings were grown for 7 weeks in nutrient solutions at pH 3.2, 4.0 or 5.0 containing 0, 100 or 400 µ M A1. At the end of this period, seedling growth, the cation exchange capacity of the roots and the amount of exchangeable Ca and Mg in roots were determined. A1 concentrations in whole roots, root segments, and in needles were measured. Using X‐ray microanalysis, the concentrations of Al, Ca, Mg and P were determined in cortical cell walls. We wanted to test the hypotheses that (1) the amount of Al bound to cation exchange sites can be used as a marker for Al toxicity and (2) the Mg concentration of needles is controlled by the amount of Mg bound to cation exchange sites. Low pH reduced the inhibition of Al on root growth and shoot length. Both low pH and Al lowered the concentration of Ca and Mg in needles. Al concentrations in the roots decreased as the pH decreased. In the roots, Al displaced Mg and Ca from binding sites at the root cortical cell walls. A comparison of the effects of Al at the different pH values on root growth and Mg concentration in the needles, suggests that, at pH 5.0, an Al fraction in the apoplast inhibits root growth, but does not affect Mg uptake. This fraction of Al is not available for transport to the shoots. In contrast, Mg uptake is strongly affected by Al at pH 3.2, although only very low levels of Al were detected in the roots. Thus, Al accumulation in the apoplast is a positive marker for Al effects on root growth, but not Mg uptake. The Mg concentration of needles is not controlled by the amount of Mg bound to cation exchange sites.     

Effects of inorganic nitrogen application on the dynamics of the soil solution composition in the root zone of maize

The effect of inorganic nitrogen (N) fertilizer on the ionic composition of the soil solution under maize (Zea mays L.) was studied. A pot experiment was carried out with two treatments combined factorially, with or without N application (Ca(NO₃)₂; +N and –N treatments, respectively), and with or without plants. Three looped hollow fiber samplers were installed in each pot to sample soil solutions nondestructively from the root zone, seven times during the 50-day growth period. Plants were harvested on the 50th day, and their nutrient contents determined.Effects of N fertilizer on the soil solutions were observed by the first sampling, 2 days after sowing. The concentrations of Ca and NO₃ ^– and electrical conductivity (EC) increased significantly in the +N treatments as direct effects of fertilizer application. In addition, the concentrations of Mg, K, Na and H⁺ also increased and that of P decreased significantly as indirect effects caused by the re-establishment of chemical equilibria. This suggested the greater supply as well as the greater possibility of leaching loss not only of NO₃ ^– but also of Ca, Mg and K. In the treatments with plants, the concentrations of NO₃ ^–, Ca, Mg and K decreased with time and pH increased significantly compared with the unplanted soil. The depletion of N in the soil solution roughly agreed with the amount of N taken up by the plant. The depletions of K from the soil solution amounted to less than 10% of the amount of the K taken up, suggesting intensive replenishment of K from exchange sites in the soil. Depletions of Ca and Mg were several times higher than the amounts taken up, indicating that the depletions resulted from the adsorption of the divalent cations by the soil rather than uptake by plants. Because NO₃ ^– is hardly absorbed by exchange sites in soil and was the dominant anion in solution, it was concluded that NO₃ ^– had a major role in controlling cation concentrations in the soil solution and, consequently, on their availability for uptake by plants as well as their possible leaching loss. ei]H Marschner     

Physicochemical selectivity for cations and CEC of grass roots

Summary A study of root selective adsorption of Ca, Mg, K and Na was made with rootlets of 20 species of Gramineae (from natural vegetation) exhibiting low cation exchange capacities (CEC) scattered over a narrow range 7.6–16.7 meq 100 g^-1 dry matter). For each sample of rootlets successive measurements were made of its CEC (amount of Ca adsorbed from 0.05 N calcium nitrate solution) and the amounts of Ca, Mg, K and Na adsorbed from a tetracationic nitrate solution having equal concentrations (0.05 N) of each cation. Rootlet selectivity for bivalent cations increased with CEC and was greater for Ca than for Mg. In contrast, the selectivity for monovalent cations increased with decreasing CEC and was greater for K than for Na. A root's selective adsorption may therefore be governed by its CEC, in conformation with the exchange laws, as is the case with clays and resins. Paper read at the XII International Botanical Congress (mineral nutrition session) held in Leningrad, July, 1975.     

Influence of alpine plants on soil nutrient concentrations in a monoculture experiment

The ability of different alpine species to influence soil nutrient concentrations was quantified by growing monocultures of 17 species on a homogenized acid alpine soil mixture. The experiment was carried out at 2750 m a.s.l. in the Teberda Reserve, Northwest Caucasus. Soil nuturient contents (NH₄, NO₃, P, Ca, Mg, and K) and pH were analyzed after 6 years. The same soil mixture but without plants was used as a control. The plant species had significant effects on all soil properties. Different species groups tended to decrease different nutrients to different extents, e.g.Matricaria caucasica had the lowest level for NO₃ andFestuca ovina for P. Many species increased the cation content (Ca, Mg, K) in the soil in comparison with the control. Prevention of cation leaching seems to be the main mechanism of these increases, because initial cation contents were higher than the final. All species, exceptSibbaldia procumbens, increased soil pH in comparison with the final control. Significant differences among taxonomic groups (families) were found for exchangeable Ca, Mg, and pH.Fabaceae decreased cation contents (Ca, Mg), but tended to increase nitrogen (NH₄, NO₃).Cyperaceae (Carex spp.) tended to decrease ammonium content, and bothAsteraceae andCyperaceae tended to decrease nitrate concentrations. The phosphorus content tended to be reduced by grasses. There was no strong correspondence between properties of native soils of 4 alpine communities and nutrient concentrations for species preferring those communities.     

Influences of different nitrate to ammonium ratios on chlorosis,cation concentrations and the binding forms of Mg and Ca in needles of Mg-deficient Norway spruce

Effects of Mg deficiency and variations of the NO₃ ^-/NH₄ ⁺-ratio on chlorophyll, mineral nutrient concentrations and the binding forms of Mg and Ca were investigated in current-year, one- and two-year-old needles of clonal Norway spruce (Picea abies [L.] Karst.). Six-year-old spruce plants were grown for one year in sand culture with circulating nutrient solutions containing sufficient (0.2 mt M) or limiting (0.04 mt M) concentrations of Mg. The NO₃ ^-/NH₄ ⁺-ratio in the nutrient solutions administered to the experimental trees was adjusted to 0.76 in the Mg-sufficient treatment and to 1.86, 0.76 and 0.035 in Mg-limited treatments. Mg and chlorophyll concentrations, were strongly influenced by the applied nitrogen source in current-year needles and - to a less extent - also in one-year-old needles. NH₄ ⁺-dominated nutrition resulted in decreased height growth and significantly lower Mg and chloropyhll concentrations in current-year and one-year-old needles compared to NO₃ ^--dominated nutrition. Decreases in total Mg were linearly correlated to reductions of water-soluble Mg and water-unsoluble Mg not bound to chlorophyll. Mg bound to chlorophyll, however, was only reduced, when total Mg decreased below a physiological threshold value of 2% of the total nitrogen concentration in the respective needles. Total Ca concentrations in the needles, which were reduced by Mg deficiency especially when nutrition was NH₄ ⁺-dominated, were strongly correlated to the portion of Ca bound to oxalate. The amount of water-soluble Ca and pectate-bound Ca remained nearly constant, independent from changes of total Ca concentrations. Negative effects of increasing NH₄ ⁺ supply on concentrations of Mg and other cations in the needles can be attributed to an inhibition of cation uptake induced by ion antagonism and/or reduced root growth.     

Nutrient fluxes in a snow-dominated, semi-arid forest: Spatial and temporal patterns

We tested five hypotheses regarding the potential effects of precipitation change on spatial and temporal patterns of water flux, ion flux, and ion concentration in a semiarid, snowmelt-dominated forest in Little Valley, Nevada. Variations in data collected from 1995 to 1999 were used to examine the potential effects of snowpack amount and duration on ion concentrations and fluxes. Soil solution NO₃ ^–, NH₄ ⁺, and ortho-phosphate concentrations and fluxes were uniformly low, and the variations in concentration bore no relationship to snowmelt water flux inputs of these ions. Weathering and cation exchange largely controlled the concentrations and fluxes of base cations from soils in these systems; however, soil solution base cation concentrations were affected by cation concentrations during snowmelt episodes. Soil solution Cl^– and SO₄ ^2– concentrations closely followed the patterns in snowmelt water, suggesting minimal buffering of either ion by soils. In contrast to other studies, the highest concentration and the majority of ion flux from the snowpack in Little Valley occurred in the later phases of snowmelt. Possible reasons for this include sublimation of the snowpack and dry deposition of organic matter during the later stages of snowmelt. Our comparison of interannual and spatial patterns revealed that variation in ion concentration rather than water flux is the most important driver of variation in ion flux. Thus, it is not safe to assume that changes in total precipitation amount will cause concomitant changes in ion inputs to this system.     

Modification of the response of saline stressed tomato plants by the correction of cation disorders

The interactions between NaCl and other major nutrients have been generally observed in plants. Decreases of nutrient uptake under saline conditions normally appear in tomato plants grown under saline conditions. In this work, the effect of increased external Ca, K and Mg concentrations under saline conditions has been investigated. Tomato plants (Lycopersicon esculentum, Mill) were grown in a greenhouse, in 120 L capacity containers, filled with continuously aerated Hoagland nutrient solution. Treatments were added to observe the combined effect of two NaCl levels (30 and 60 mM) and three levels of Ca, K and Mg (in mM ratios of 4:6:1, 7:9:2 and 10:12:3; treatments C₁, C₂ and C₃ respectively) on growth, fruit yield and water relations. Saline treatments decreased the growth, which was partly restored with the C₂ treatment and totally with the C₃ treatment. A good association was observed between the electric conductivity of the medium and the water or osmotic potential of the leaves, independent of the type of treatment (salinity or cation ratio). Salinity at 30 and 60 mM NaCl reduced the fruit yield compared with that obtained at 0 mM NaCl. However, there was an increase, as a consequence of the application of treatments C₂ and C₃, in each saline treatment. At a high salinity level (60 mM), the ratios Na/K, Na/Ca and Na/Mg in young leaves decreased as a consequence of cation treatments. Higher concentrations of sugars in leaves and fruits were obtained after increasing the salinity and cation concentrations. Also, sucrose phosphate synthase activity in leaves and fruits was increased after the treatments, but there was no measurable invertase activity in fruits. Therefore, the concentrations of Ca, K and Mg in the nutrient solution could be important factors in the hydroponic culture of tomato grown under saline conditions.     

Equilibrium and Ion Exchange Characteristics of Potassium and Sodium Accumulation by Barley Roots

Potassium ion and Na⁺ influx and efflux rates into and from excised barley roots are compared with the maximum capacity of accumulation. Potassium ion and Na⁺ influx and efflux involve a cation exchange that is independent of simultaneous exchange of the accompanying anion. These exchange fluxes depend on the concentration and cation composition of the solutions from which they originate. Selective differences between K⁺ and Na⁺ fluxes are sufficient to account for a cationic distribution within the roots that differs markedly from that of the external solution and that persists for extended time periods. The accumulation maximum is a cation exchange equilibrium with the cation influx and efflux rates approaching equality. The equilibrium level is independent of the individual cation fluxes and the external solution concentration. It is a finite quantity which appears to be determined by the internal anion concentration including accumulated as well as endogenous anions.     

Nutrient retention by Pseudoscleropodium purum and its relation to growth

Abstract

A complete mineral nutrient fertilizer solution was added to carpets of Pseudoseleropodium purum growing in Windsor Forest, Berkshire, to study the efficiency of nutrient capture from dilute solutions and its effect on growth. In the experiment different frequencies of application were balanced by employing different concentrations of fertilizer so that all treatment plots received the same quantities of nutrients over a six-month period. At the end of the experiment the concentrations of macronutrients (N, P, K, Mg, Ca) within the moss tissues were determined and shoot growth was assessed by the tagged shoot method.

Significant luxury uptake occurred only in the case of P where shoot concentrations in fertilized plots were 50% above the control. Smaller net increases were noted for other elements, particularly Mg. Shoot N concentration was hardly affected by fertilizer application possibly due to efficient internal recycling of the element in P. purum. Levels of exchange ably held cations were not markedly increased by fertilizer addition in the field experiment but when shoots were incubated with fertilizers in the laboratory for 30 min the concentrations of exchangeable Ca and Mg rose appreciably and exchangeable K fell. These changes appear to be quickly reversed under field conditions without transfer of cations to the protoplasts. The importance of elevated cation exchange capacity in the initial sequestration of nutrient cations is, therefore, questioned. Growth stimulation due to fertilizer treatment was negligible. Maximum net uptake of P, Mg and Ca occurred when plots were watered at weekly intervals with a dilute fertilizer and least occurred where a concentrated solution was applied at less frequent intervals implying that contact time between P. purum and its potential nutrients is an important factor. The period elapsing between fertilizer additions may also have significance, however, due to leaching away of nutrients and the re-establishment of a natural ionic balance at the cell wall exchange sites. Levels of nutrient absorption and growth were greatest at a site where the moss remained moist longest. This result indicates that the ability of P. purum to absorb and utilize additional nutrients is governed by general metabolic performance which is itself restricted by water supply in Windsor Forest.     

Macro nutrient cation uptake by plants

Summary In pot experiments with barley, mustard, leek, lettuce and spinach, and in a field experiment with 30 cultivars of barley uptakes of K, Mg, Ca, Na and N were studied at varying concentrations and activities of these cations in the soil solution.The sum of macro cations (K, Mg, Ca, Na) in meq per 100 g aerial plant parts were independent of the chemical composition of the soil solution, but dependent on plant species and on the N concentration in the plant.The ratios of mean net inflows of Mg, Ca and K into plants and corresponding cation activity ratios (a_Mg/a_Ca and ) in the soil solution were linearly related and highly correlated under conditions in which growth rate and/or rate of incorporation into new tissues constituted the rate determining step of cation uptake. Consequently, mean net inflows of K, Mg and Ca were independent of ion concentration and ion activity of K, Mg or Ca in the soil solution under the conditions of constant activity ratio.The results agree with the concept that plants have a finite cation uptake capacity, and that plants are in a equilibrium-like state with the activities of K, Mg, and Ca ions in the soil solution. The results indicate that both ratios and content of exchangeable cations should be considered in our evaluation of soil test data.     

Salt or ion bridges in biological systems: a study employing quantum and molecular mechanics

Equilibrium geometries and binding energies of model "salt" or "ion" bridge systems have been computed by ab initio quantum chemistry techniques (GAUSSIAN82) and by empirical force field techniques (AMBER2.0). Formate and dimethyl phosphate served as anions in the model compounds while interacting with several organic cations, including methyl ammonium, methyl guanidinium, and divalent metal ion (either Mg2+ or Ca2+) without and with an additional chloride; and a divalent metal ion (either Mg2+ or Ca2+), chloride, and four water molecules of hydration about the metal ion. The majority of the quantum chemical computations were performed using a split-valence basis set. For the model compounds studied we find that the ab initio optimized geometries are in remarkably good agreement with the molecular mechanics geometries. Several calculations were also performed using diffuse fractions. The formate anion binds these model cations more strongly than does dimethyl phosphate, while the organic cation methyl ammonium binds model anions more strongly than does methyl guanidinium. Finally, in model compounds including organic anions, Mg2+ or Ca2+ and four molecules of water, and a chloride anion, we find that the equilibrium structure of the magnesium complex involves a solvent separated ion pair (the magnesium ion is six coordinate), whereas the calcium ion complex remains seven coordinate. Molecular mechanics overestimates binding energies, but the estimates may be close enough to actual binding energies to give useful insight into the details of salt bridges in biological systems.     

Systematic Separation of Human Urinary Peptides

Urinary peptides were roughly fractionated by combined columns of cation and anion exchange resins, and the peptides eluted from each column were further fractionated by a combination of various ion exchange resins and DEAE-cellulose column chromatography, paper chromatography and other methods. From the fractions adsorbed on cation exchange resin, 13 homogeneous peptides could be isolated, and from the ones adsorbed on anion exchange resin, 8 glycopeptides could be found. Their amino acid compositions were analyzed.

Although some fractions remain univestigated, an outline of the whole aspect of main urinary peptides has been clarified by this study.     

Transport of cations and anions across forestomach epithelia: conclusions from in vitro studies

Secretion of saliva as well as absorptive and secretory processes across forestomach epithelia ensures an optimal environment for microbial digestion in the forestomachs. Daily salivary secretion of sodium (Na+) exceeds the amount found in plasma by a factor of 2 to 3, while the secretion of bicarbonate (HCO3-) is 6 to 8 times higher than the amount of HCO3- in the total extracellular space. This implies a need for efficient absorptive mechanisms across forestomach epithelia to allow for an early recycling. While Na+ is absorbed from all forestomachs via Na+/H+ exchange and a non-selective cation channel that shows increased conductance at low concentrations of Mg2+, Ca2+ or H+ in the luminal microclima and at low intracellular Mg2+, HCO3- is secreted by the rumen for the buffering of ingesta but absorbed by the omasum to prevent liberation of CO2 in the abomasum. Fermentation provides short chain fatty acids and ammonia (NH3) that have to be absorbed both to meet nutrient requirements and maintain ruminal homeostasis of pH and osmolarity. The rumen is an important location for the absorption of essential minerals such as Mg2+ from the diet. Other ions can be absorbed, if delivered in sufficient amounts (Ca2+, Pi, K+, Cl- and NH4+). Although the presence of transport mechanisms for these electrolytes has been described earlier, our knowledge about their nature, regulation and crosstalk has increased greatly in the last years. New transport pathways have recently been added to our picture of epithelial transport across rumen and omasum, including an apical non-selective cation conductance, a basolateral anion conductance, an apical H+-ATPase, differently expressed anion exchangers and monocarboxylate transporters.     

Tight binding of divalent cations to monomeric actin. Binding kinetics support a simplified model

Using the fluorescent Ca2+ selective chelator Quin2 to induce and measure the dissociation of Ca2+ from actin, we have recently found that actin binds Ca2+ and Mg2+ much more tightly than previously thought (Gershman, L.C., Selden, L.A., and Estes, J.E. (1986) Biochem. Biophys. Res. Commun. 135, 607-614). In this report, we show that the kinetics of dissociation of Ca2+ from Ca-actin and Mg2+ from Mg-actin closely parallel the fluorescence changes in 1,5-I-N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine (AEDANS)-actin, suggesting that the 1,5-I-AEDANS-actin fluorescence directly reflects slow first-order cation exchange rather than a slow Mg2+-induced isomerization as originally proposed by Frieden (Frieden, C. (1982) J. Biol. Chem. 257, 2882-2886). Measuring divalent cation exchange directly, we have determined the dissociation rate constants for Ca2+ (k-Ca) and Mg2+ (k-Mg), the equilibrium dissociation constants for Ca2+ (KCa), and the ratio of cation binding affinities, KMg/Kca, to actin over the pH range 7-8. We have found that k-Ca is 5-10 times greater than k-Mg and KMg is about 4 times greater than KCa. From the data we calculate the association rate constants for Ca2+ (kCa) and Mg2+ (kMg) to be about 7 X 10(6) M-1 s-1 and 2 X 10(5) M-1 s-1, respectively. kCa appears to be diffusion-limited, but kMg is significantly smaller due to the characteristics of the Mg2+ aquo ion. These findings are consistent with a simple first-order binding model for the tight binding of divalent cations to actin.     

The binding of Ca2+ to solubilized band 3 protein of the human erythrocyte membrane

The binding of 45Ca2+ to isolated band 3 protein, the anion transport protein of the human erythrocyte membrane, was studied by equilibrium dialysis. The protein was solubilized and purified by either the nonionic detergent Ammonyx-L0 or acetic acid. Each preparation of band 3 protein showed a single high-affinity Ca2+ binding site and several Ca2+ binding sites of lower affinity. The association constant of the high-affinity site was 4-13 X 10(4)M-1; it was only moderately dependent on ionic strength. Mg2+ effectively competed with Ca2+ for the site. Anion exchange across the human erythrocyte membrane is inhibited by micromolar concentrations of intracellular Ca2+. Our results suggest that this inhibition is due to the binding of the cation to a single site on band 3 protein.