Calcium Binding Capacity Research Articles

Oxalate content and calcium binding capacity of tea and herbal teas.

Thirty-two commercially available teas consisting of green, oolong and black teas were bought from supermarkets in Christchurch, New Zealand in June 2001. Fifteen herbal teas were also purchased at the same time. The soluble oxalate content of the infusate made from each of the teas was determined using high pressure liquid chromatography. The mean soluble oxalate contents of black tea in tea bags and loose tea leaves were 4.68 and 5.11 mg/g tea, respectively, while green teas and oolong tea had lower oxalate contents, ranging from 0.23 to 1.15 mg/g tea. The soluble oxalate content of the herbal teas ranged from not detected to 3.00 mg/g tea. A regular tea drinker consuming six cups of tea/day would have an intake of between 26.46 and 98.58 mg soluble oxalate/day from loose black tea, 17.88 and 93.66 mg soluble oxalate/day from black tea in tea bags and a maximum of 18.0 mg/day from herbal teas. The oxalate intake from the regular daily consumption of black teas is modest when compared to the amounts of soluble oxalate that can be found in common foods. However, oxalate in black teas has the potential to bind to a significant proportion of calcium in the milk, which is commonly consumed with the black teas.

Crystallization of δ-Na2Si2O5-rich layered silicates from sodium silicate solutions: seeding and temperature programmed δ-phase embryo creation

Abstract The effect of seed composition on the crystalline layered sodium disilicate phase development using a sodium silicate solution (SiO 2 /Na 2 O molar ratio=2.1) has been investigated. To obtain δ phase-rich products, it is necessary that the α phase content in the seed be as low as possible. High temperatures (700 °C) and absence of any α phase in the initial seed substantially promote the amorphous phase→δ phase transformation reaction even if the seed contains up to 15.0 wt.% β phase. It has been shown that pretreatment in the temperature range 150–200 °C promotes significantly δ phase nucleation. Low temperature nucleation has been observed for the sodium silicate solution modulus in the range of 1.5–2.1. This finding might be implemented to solve reproducibility problems in the industrial scale production of layered disilicates from water glass. To obtain a minimum calcium binding capacity of 120.0 mg CaO/g , an α phase content of up to 20.0 wt.% is tolerable.

Effect of particle sizes on in-vitro calcium and magnesium binding capacity of prepared dietary fibers

Microscopic examination showed a change on the surface structure of dietary fibers (DF) from rice straw (RS) and sugarcane bagasse (SB) by alkaline hydrogen peroxide (AHP) treatment. Effect of particle size on in-vitro binding of calcium (Ca) and magnesium (Mg) of AHP-RS, AHP-SB and Solka Floc® 900 (SF) were evaluated. Total bound Ca and Mg decreased with particle size reduction ( P<0.05) for all types of DF. Ca and Mg (82–95%) were released from AHP-RS and AHP-SB by acid washing but only 53–60% Mg was removed from SF. There were strong correlations between water holding capacity, oil binding capacity and particle density and total bound Ca and Mg for all DF samples.

Steroleosin, a Sterol-Binding Dehydrogenase in Seed Oil Bodies

Abstract Besides abundant oleosin, three minor proteins, Sop 1, 2, and 3, are present in sesame (Sesamum indicum) oil bodies. The gene encoding Sop1, named caleosin for its calcium-binding capacity, has recently been cloned. In this study, Sop2 gene was obtained by immunoscreening, and it was subsequently confirmed by amino acid partial sequencing and immunological recognition of its overexpressed protein in Escherichia coli. Immunological cross recognition implies that Sop2 exists in seed oil bodies of diverse species. Along with oleosin and caleosin genes, Sop2 gene was transcribed in maturing seeds where oil bodies are actively assembled. Sequence analysis reveals that Sop2, tentatively named steroleosin, possesses a hydrophobic anchoring segment preceding a soluble domain homologous to sterol-binding dehydrogenases/reductases involved in signal transduction in diverse organisms. Three-dimensional structure of the soluble domain was predicted via homology modeling. The structure forms a seven-stranded parallel β-sheet with the active site, S-(12X)-Y-(3X)-K, between an NADPH and a sterol-binding subdomain. Sterol-coupling dehydrogenase activity was demonstrated in the overexpressed soluble domain of steroleosin as well as in purified oil bodies. Southern hybridization suggests that one steroleosin gene and certain homologous genes may be present in the sesame genome. Comparably, eight hypothetical steroleosin-like proteins are present in the Arabidopsis genome with a conserved NADPH-binding subdomain, but a divergent sterol-binding subdomain. It is indicated that steroleosin-like proteins may represent a class of dehydrogenases/reductases that are involved in plant signal transduction regulated by various sterols.

CABYR, a Novel Calcium-Binding Tyrosine Phosphorylation-Regulated Fibrous Sheath Protein Involved in Capacitation

To reach fertilization competence, sperm undergo an incompletely understood series of morphological and molecular maturational processes, termed capacitation, involving, among other processes, protein tyrosine phosphorylation and increased intracellular calcium. Hyperactivated motility and an ability to undergo the acrosome reaction serve as physiological end points to assess successful capacitation. We report here that acidic (pI 4.0) 86-kDa isoforms of a novel, polymorphic, testis-specific protein, designated calcium-binding tyrosine phosphorylation-regulated protein (CABYR), were tyrosine phosphorylated during in vitro capacitation and bound 45Ca on 2D gels. Acidic 86-kDa calcium-binding forms of CABYR increased during in vitro capacitation, and calcium binding to these acidic forms was abolished by dephosphorylation with alkaline phosphatase. Six variants of CABYR containing two coding regions (CR-A and CR-B) were cloned from human testis cDNA libraries, including five variants with alternative splice deletions. A motif homologous to the RII dimerization domain of PK-A was present in the N-terminus of CR-A in four CABYR variants. A single putative EF handlike motif was noted in CR-A at aas 197–209, while seven potential tyrosine phosphorylation-like sites were noted in CR-A and four in CR-B. Pro-X-X-Pro (PXXP) modules were identified in the N- and C-termini of CR-A and CR-B. CABYR localizes to the principal piece of the human sperm flagellum in association with the fibrous sheath and is the first demonstration of a sperm protein that gains calcium-binding capacity when phosphorylated during capacitation.

Synthesis and Characterization of Detergent-Grade Zeolite from Indian Clay

Detergent-grade zeolite 4A has been synthesized from the clays found in various parts of India. The process parameters for the synthesis of detergent-grade zeolite, like the activation temperature for the chemical transformation of clay, the composition of the reacting mixture and finally the crystallization temperature, have been studied. Better metakaolinite formation has been observed at a calcination temperature of around 550°C. The characterization of the synthesized zeolite by X-ray diffraction study confirms its formation. Moreover, the calcium binding capacity 170 mg of CaO/gm as well as particle size in the range of 3-7 microns of this zeolite are found to be satisfactory characteristics to substitute phosphate in detergents.

Highly crystalline zeolite — a from flyash of bituminous and lignite coal combustion

Flyash is being generated in voluminous amounts by large scale coal combustion process. It poses a serious threat to thermal power industries specifically, in India, wherein the percent of utilisation of flyash is very poor (3–5%). In view of this problem, newer methods of its disposal and utilisation are being explored. The synthesis of zeolite from flyash appears to be one of the most promising alternatives as it has emphasis on value addition to waste material. Flyashes originating from different sources of coal differ in their characteristics and have implications in this work on Zeolite-A production. These factors have been thoroughly investigated and the conditions favourable for formation of Zeolite-A have been delineated. The reactivity of flyash towards zeolite formation is directly dependent on the SiO 2/Al 2O 3 ratio, Fe 2O 3 and CaO content. Amongst the flyashes investigated, so far the sub-bituminous coal based flyash with SiO 2/Al 2O 3 ratio of 3.47 appears to be a suitable substrate for Zeolite-A synthesis. These zeolites have been characterised with respect to XRD crystallinity, calcium binding capacity (CBC) and sorption capacity, wherein the crystallinity ranges from 50 to 100%, the CBC ranges from 290 to 560 meq/100 g and sorption capacity ranges from 16.6 to 23.8%.

Calcium-binding capacities of different brans under simulated gastrointestinal pH conditions. In vitro study with (45)Ca.

The present study was performed to investigate calcium-binding characteristics of different brans under simulated gastrointestinal pH conditions and to explore the significance of dietary fiber, oxalate, and phytate for calcium binding. Different brans (rice, rye, soy, fine wheat, coarse wheat, and oat) and CaCl(2) solution containing (45)Ca were incubated at 37 degrees C at gastric pH (2.2) followed by buffering steps of 1 degree from pH 3.0 to pH 8.0. Total calcium binding and calcium-binding capacity of the pH 2.2 soluble bran fraction were determined. Additionally, oxalate and phytate contents of brans and solubility profiles of phytic acid were investigated. Calcium-binding capacities of brans showed a clear pH dependence. At gastric pH calcium binding was low in all brans, ranging from 0.022 to 0.040 mmol of calcium/g of bran. Soy bran, nearly phytate-free, showed higher binding values up to pH 4.0 and lower values between pH 5.0 and 8.0. In all other brans, binding values increased strongly with increasing pH in the quantitative order rice bran > coarse wheat bran > fine wheat bran > rye bran > oat bran. The solubility profiles indicate that in the cases of rye, wheat, and rice bran phytate accounts for 70-82% of their total calcium-binding capacities. The results suggest that dietary fiber makes no important contribution to calcium binding, except for soy and oat brans. Oxalate plays only a minor role in calcium binding by brans.

An MCell model of calcium dynamics and frequency-dependence of calmodulin activation in dendritic spines

Pairing action potentials in synaptically coupled cortical pyramidal cells induces LTP in a frequency-dependent manner (H. Markram et al., Science 275 (1997) 213). Using MCell, which simulated the 3D geometry of the spine and the diffusion and binding of Ca 2+, we show that pairing five EPSPs and back-propagating action potentials results in a Ca 2+ influx into a model dendritic spine that is largely frequency independent but leads to a frequency-dependent activation of postsynaptic calmodulin. Furthermore, we show how altering the availability of calmodulin and the calcium-binding capacity can alter the efficacy and potency of the frequency-response curve. The model shows how the cell can regulate its plasticity by buffering Ca 2+ signals.

Altered middle lamella homogalacturonan and disrupted deposition of (1-->5)-alpha-L-arabinan in the pericarp of Cnr, a ripening mutant of tomato.

Cnr (colorless non-ripening) is a pleiotropic tomato (Lycopersicon esculentum) fruit ripening mutant with altered tissue properties including weaker cell-to-cell contacts in the pericarp (A.J. Thompson, M. Tor, C.S. Barry, J. Vrebalov, C. Orfila, M.C. Jarvis, J.J. Giovannoni, D. Grierson, G.B. Seymour [1999] Plant Physiol 120: 383-390). Whereas the genetic basis of the Cnr mutation is being identified by molecular analyses, here we report the identification of cell biological factors underlying the Cnr texture phenotype. In comparison with wild type, ripe-stage Cnr fruits have stronger, non-swollen cell walls (CW) throughout the pericarp and extensive intercellular space in the inner pericarp. Using electron energy loss spectroscopy imaging of calcium-binding capacity and anti-homogalacturonan (HG) antibody probes (PAM1 and JIM5) we demonstrate that maturation processes involving middle lamella HG are altered in Cnr fruit, resulting in the absence or a low level of HG-/calcium-based cell adhesion. We also demonstrate that the deposition of (1-->5)-alpha-L-arabinan is disrupted in Cnr pericarp CW and that this disruption occurs prior to fruit ripening. The relationship between the disruption of (1-->5)-alpha-L-arabinan deposition in pericarp CW and the Cnr phenotype is discussed.

A calreticulin-like molecule from the human hookworm Necator americanus interacts with C1q and the cytoplasmic signalling domains of some integrins.

Calreticulin was recently identified as a hookworm (Necator americanus) allergen, implying secretion, and contact with cells of the immune system, or significant worm attrition in the tissues of the host. As human calreticulin has been shown to bind to and neutralize the haemolytic activity of the complement component C1q, and to be putatively involved in integrin-mediated intracellular signalling events in platelets, it was of interest to determine whether a calreticulin from a successful nematode parasite of humans, with known immune modulatory and antihaemostatic properties, exhibited a capacity to interfere with complement activation and to interact with integrin domains associated with cell signalling in platelets and other leucocytes. We can now report that recombinant calreticulin failed to demonstrate significant calcium binding capacity, which is a hallmark of calreticulins in general and may indicate inappropriate folding following expression in a prokaryote. Nevertheless, recombinant calreticulin retained sufficient molecular architecture to bind to, and inhibit the haemolytic capacity of, human C1q. Furthermore, recombinant calreticulin reacted in surface plasmon resonance analysis (SPR) with peptides corresponding to cytoplasmic signalling domains of the integrins alphaIIb and alpha5, in a calcium independent manner. SPR was also used to ratify the specificity of a polyclonal antibody to hookworm calreticulin, which was then used to assess the stage specificity of expression of the native molecule (in comparison with reverse transcriptase-polymerase chain reaction), to indicate its apparent secretion, and to purify native calreticulin from worm extracts by affinity chromatography. This development will allow the functional tests described above to be repeated for native calreticulin, to ascertain its role in the host-parasite relationship.

Structural basis for low catalytic activity in Lys49 phospholipases A2--a hypothesis: the crystal structure of piratoxin II complexed to fatty acid.

Asp49 plays a fundamental role in supporting catalysis by phospholipases A2 by coordinating the calcium ion which aids in the stabilization of the tetrahedral intermediate. In several myotoxins from the venoms of Viperidae snakes, this aspartic acid is substituted by lysine. The loss of calcium binding capacity by these mutants has become regarded as the standard explanation for their greatly reduced or nonexistent phospholipolytic activity. Here we describe the crystal structure of one such Lys49 PLA2, piratoxin-II, in which a fatty acid molecule is observed within the substrate channel. This suggests that such toxins may be active enzymes in which catalysis is interrupted at the stage of substrate release. Comparison of the present structure with other PLA2s, both active and inactive, identifies Lys122 as one of the likely causes of the increased affinity for fatty acid in Lys49 enzymes. Its interaction with the mainchain carbonyl of Cys29 is expected to lead to hyperpolarization of the peptide bond between residues 29 and 30 leading to an increased affinity for the fatty acid headgroup. This strongly bound fatty acid may serve as an anchor to secure the toxin within the membrane thus facilitating its pathological effects.

Preparation of novel functional oligophosphopeptides from hen egg yolk phosvitin.

Novel hen egg phosvitin phosphopeptides (PPP) with molecular masses of 1-3 kDa were prepared from tryptic hydrolysis following partial alkaline dephosphorylation. The phosvitin treated with various NaOH concentrations (0.05-0.4 N) resulted in different dephosphorylated proteins ranging from 17.5 to 96.3% of phosphate retention. The protein was digested into 10-20 amino acid peptides with trypsin. Calcium-binding properties of PPP were compared with those of commercial casein phosphopeptides in vitro. The PPP with 35% phosphate retention is shown to be effective for enhancing calcium binding capacity and inhibiting the formation of insoluble calcium phosphate. The results provide potential novel functional oilgophosphopeptides as nutraceuticals.

Altered expression of calcium- and apoptosis-regulating proteins in multiple system atrophy Purkinje cells.

The expression patterns of the calcium binding proteins calbindin and parvalbumin and of the apoptosis modulating proteins Bcl-2, Bax, and Bcl-x were studied in the cerebellum of patients with multiple system atrophy (MSA). Calbindin and parvalbumin immunoreactivity was markedly decreased in MSA Purkinje cells whereas Bax and Bcl-x protein expression was increased. Bcl-2 expression was restricted to a subpopulation of granule neurons, but no decrease of Bcl-2 was evident in MSA. Additional DNA end-labeling (ISEL) studies revealed only one possible apoptotic Purkinje cell nucleus, but nuclei in the cerebellar white matter, probably oligodendrocytes, in the cerebellum of patients with MSA. The present results suggest that a diminished calcium binding capacity of MSA Purkinje cells might lead to a change in the regulation of proteins of the bcl-2 family that could favor the pathologic initiation of apoptosis.

Ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) inhibits the growth of Escherichia coli at alkaline pH.

The growth of Escherichia coli was inhibited by ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) in a medium of initial pH 8.8. The growth inhibition was reversed by the addition of CaCl(2). E. coli could grow in the presence of EGTA at pH values below 8. The concentration of free calcium ions increases with a decrease in medium pH because of a decrease in the calcium binding capacity of EGTA. So, although the results suggest that calcium ions are essential for the growth of E. coli, the minimum concentration required is very low.

Activity-Related Calcium Dynamics in Motoneurons of the Nucleus Hypoglossus From Mouse

A quantitative analysis of activity-related calcium dynamics was performed in motoneurons of the nucleus hypoglossus in the brain stem slice preparation from mouse by simultaneous patch-clamp and microfluorometric calcium measurements. Motoneurons were analyzed under in vitro conditions that kept them in a functionally intact state represented by rhythmic, inspiratory-related bursts of excitatory postsynaptic currents and associated action potential discharges. Bursts of electrical activity were paralleled by somatic calcium transients resulting from calcium influx through voltage-activated calcium channels, where each action potential accounted for a calcium-mediated charge influx around 2 pC into the somatic compartment. Under in vivo conditions, rhythmic-respiratory activity in young mice occurred at frequencies up to 5 Hz, demonstrating the necessity for rapid calcium elevation and recovery in respiratory-related neurons. The quantitative analysis of hypoglossal calcium homeostasis identified an average extrusion rate, but an exceptionally low endogenous calcium binding capacity as cellular parameters accounting for rapid calcium signaling. Our results suggest that dynamics of somatic calcium transients 1) define an upper limit for the maximum frequency of respiratory-related burst discharges and 2) represent a potentially dangerous determinant of intracellular calcium profiles during pathophysiological and/or excitotoxic conditions.

Calcium binding capacity of the cytosol and endoplasmic reticulum of mouse pancreatic acinar cells.

1. The droplet technique was used in this study to measure total calcium loss from pancreatic acinar cells due to calcium extrusion. The calcium binding capacity of the cytosol (kc) was measured as the ratio of the decrease in the total calcium concentration of the cytosol of the cell (Delta[Ca]c) and the synchronously occurring decrease in the free calcium ion concentration in the cytosol (Delta[Ca2+]c). The calcium dependency of the calcium binding capacity was determined by plotting values of kc against the corresponding [Ca2+]c. The rise in the cytosolic Ca2+ concentration of pancreatic acinar cells was triggered by stimulation with a supramaximal dose of cholecystokinin (CCK). The recovery of [Ca2+]c during continued exposure to the agonist was due to calcium extrusion from the cell. 2. The calcium binding capacity was about 1500-2000 for the [Ca2+]c range 150-500 nM. The mechanism of buffering was not investigated in this study. The calcium binding capacity of the cytosol did not vary significantly with [Ca2+]c in this range. The CCK-evoked decrease in the total calcium concentration in the lumen of the endoplasmic reticulum (ER) can be estimated from our data, taking into account previously published values for the volume of the ER in pancreatic acinar cells. Comparing the decrease in the total ER calcium concentration with our recently reported values for agonist-induced reductions in the free Ca2+ concentration inside the ER, we estimate that the calcium binding capacity of the ER is approximately 20. In pancreatic acinar cells we have therefore found a difference of two orders of magnitude in the efficiency of calcium buffering in the cytosol and the ER lumen.

Heterologous Overexpression of Human NEFA and Studies on the Two EF-Hand Calcium-Binding Sites

Human NEFA is an EF-hand, leucine zipper protein containing a signal sequence. To confirm the calcium binding capacity of NEFA, recombinant NEFA analogous to the mature protein and mutants with deletions in the EF-hand domain were expressed in Pichia pastoris and secreted into the culture medium at high yield. The calcium binding activity of each purified protein was measured by a modified equilibrium dialysis using the fluorescent Ca2+ indicator FURA-2 and atomic absorption spectroscopy. A stoichiometry of 2 mol Ca2+/mol NEFA was determined. The Ca2+ binding constants were resolved by intrinsic fluorescence spectroscopy. Fluorescence titration exhibited two classes of Ca2+ binding sites with Kd values of 0.08 μM and 0.2 μM. Circular dichroism (CD) spectroscopy showed an increase from 30 to 43% in the amount of α-helix in NEFA after addition of calcium ions. Limited proteolytic digestion indicated a Ca2+ dependent conformational change accompanied by an altered accessibility to the enzyme.

Study of calcium binding to different preparations of yeast ( Saccharomyces cerevisiae) protein by using an ion selective electrode

The calcium binding capacity of a native brewer’s yeast protein (PL) was compared to protein from the same source, after treatment with 3% sodium trimetaphosphate (PLP). Calcium titration was done by the use of an ion-selective electrode. For measurements made at pH 6.5, the number of calcium binding sites per 10 5g protein was 29 for PL and 37 for PLP. The association constant K was 3.2×10 −3 and 2.7×10 −3 for PL and PLP, respectively. Dialysis of sample PLP against 0.01 M EDTA increased the number of binding sites to 49. The higher number of binding sites in PLP is explained by the presence of phosphate moieties associated with the protein molecules and also by possible conformational changes occurring in the protein molecules as a result of STMP treatment, exposing a larger number of amino acid side chains capable of combining with Ca 2+.©

A study of strontium binding to albumins, by a chromatographic method involving atomic emission spectrometric detection

A chromatographic method involving ICP-AES (inductively coupled plasma atomic emission spectrometry) detection has been successfully applied for the study of strontium-protein complexes. The chromatographic step involves the use of gel filtration—a large-zone Hummel and Dreyer method—which allows to dissociate the bound metallic ions and the free ones. This step is followed by an ICP-AES analysis of fractions collected throughout the chromatographic experiment: the concentration of ionic metallic species in solution can therefore be calculated. Two proteins have been tested: bovine serum albumin, which showed only weak interactions with Sr 2+ ions, and bovine α-lactalbumin: this protein, well-known for its calcium binding capacity, proved to interact strongly with strontium. The influence of various parameters on the formation of strontium-lactalbumin complexes were determined, namely temperature, pH. Competition experiments between Sr 2+ ions and, respectively Na + and Ca 2+ ions were also performed, by varying ionic strength of the medium, and by using both apo and native forms of bovine α-lactalbumin.