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1.
Sci Rep ; 11(1): 19874, 2021 10 06.
Article in English | MEDLINE | ID: mdl-34615988

ABSTRACT

We report here hourly variations of Mg/Ca, Sr/Ca, and Ba/Ca ratios in a Mediterranean mussel shell (Mytilus galloprovincialis) collected at the Otsuchi bay, on the Pacific coast of northeastern Japan. This bivalve was living in the intertidal zone, where such organisms are known to form a daily or bidaily growth line comprised of abundant organic matter. Mg/Ca ratios of the inner surface of the outer shell layer, corresponding to the most recent date, show cyclic changes at 25-90 µm intervals, while no interpretable variations are observed in Sr/Ca and Ba/Ca ratios. High Mg/Ca ratios were probably established by (1) cessation of the external supply of Ca and organic layer forming when the shell is closed at low tide, and (2) the strong binding of Mg to the organic layer, but not of Sr and Ba. Immediately following the great tsunami induced by the 2011 Tohoku earthquake, Mg/Ca enrichment occurred, up to 10 times that of normal low tide, while apparent Ba/Ca enrichment was observed for only a few days following the event, therefore serving a proxy of the past tsunami. Following the tsunami, periodic peaks and troughs in Mg/Ca continued, perhaps due to a biological memory effect as an endogenous clock.


Subject(s)
Animal Shells/metabolism , Mytilus/physiology , Tsunamis , Animal Shells/chemistry , Animals , Biomarkers , Metals, Alkaline Earth/analysis , Metals, Alkaline Earth/metabolism , Spectrum Analysis , Time Factors
2.
Luminescence ; 35(2): 321-327, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31837194

ABSTRACT

Tea, originating from China, is an important part of Chinese traditional culture. There are different qualities of and producing areas for tea on the market, therefore it is necessary to discriminate between teas in a fast and accurate way. In this study, a chemical sensor array based on nanozymes was developed to discriminate between different metal ions and teas. The indicators for the sensor array are three kinds of nanozymes mimicking laccase (Cu-ATP, Cu-ADP, Cu-AMP). The as-developed sensor array successfully discriminated 12 metal ions and the detection limit was as low as 0.01 µM. The as-developed sensor array was also able to discriminate tea samples. Different kinds of tea samples appeared in different areas in the canonical score plot with different response patterns. Furthermore, in a blind experiment, we successfully discriminated 12 samples with a 100% accuracy. This sensor array integrates chemistry and food science together, realizing the simultaneous detection of several kinds of teas using a sensitive method. The as-developed sensor array would have an application in the tea market and provide a fast and easy method to discriminate between teas.


Subject(s)
Colorimetry , Laccase/metabolism , Metals, Alkaline Earth/metabolism , Metals, Heavy/metabolism , Nanoparticles/metabolism , Tea/metabolism , Laccase/chemical synthesis , Laccase/chemistry , Metals, Alkaline Earth/analysis , Metals, Heavy/analysis , Nanoparticles/chemistry , Particle Size , Surface Properties , Tea/chemistry
3.
Bioelectrochemistry ; 109: 49-56, 2016 Jun.
Article in English | MEDLINE | ID: mdl-26849106

ABSTRACT

A growing body of evidence suggests that the regulation of cardiac ryanodine receptor (RYR2) by luminal Ca(2+) is mediated by luminal binding sites located on the RYR2 channel itself and/or its auxiliary protein, calsequestrin. The localization and structure of RYR2-resident binding sites are not known because of the lack of a high-resolution structure of RYR2 luminal regions. To obtain the first structural insight, we probed the RYR2 luminal face stripped of calsequestrin by alkaline earth metal divalents (M(2+): Mg(2+), Ca(2+), Sr(2+) or Ba(2+)). We show that the RYR2 response to caffeine at the single-channel level is significantly modified by the nature of luminal M(2+). Moreover, we performed competition experiments by varying the concentration of luminal M(2+) (Mg(2+), Sr(2+) or Ba(2+)) from 8 mM to 53 mM and investigated its ability to compete with 1mM luminal Ca(2+). We demonstrate that all tested M(2+) bind to exactly the same RYR2 luminal binding sites. Their affinities decrease in the order: Ca(2+)>Sr(2+)>Mg(2+)~Ba(2+), showing a strong correlation with the M(2+) affinity of the EF-hand motif. This indicates that the RYR2 luminal binding regions and the EF-hand motif likely share some structural similarities because the structure ties directly to the function.


Subject(s)
Anticonvulsants/pharmacology , Calcium/metabolism , Metals, Alkaline Earth/metabolism , Myocardium/metabolism , Phenobarbital/pharmacology , Ryanodine Receptor Calcium Release Channel/metabolism , Amino Acid Sequence , Animals , Anticonvulsants/metabolism , Barium/metabolism , Binding Sites , Caffeine/pharmacology , Cations, Divalent/metabolism , Central Nervous System Stimulants/pharmacology , EF Hand Motifs , Humans , Magnesium/metabolism , Molecular Sequence Data , Myocardium/chemistry , Phenobarbital/metabolism , Ryanodine Receptor Calcium Release Channel/chemistry , Sequence Alignment , Strontium/metabolism
4.
J Environ Radioact ; 151 Pt 3: 551-7, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26277654

ABSTRACT

In this study, uptake of Ra from soil, and the influence of group II metals on Ra uptake, into the stones and edible flesh of the fruit of the wild green plum, Buchanania obovata, was investigated. Selective extraction of the exchangeable fraction of the soil samples was undertaken but was not shown to more reliably predict Ra uptake than total soil Ra activity concentration. Comparison of the group II metal to Ca ratios (i.e. Sr/Ca, Ba/Ca, Ra/Ca) in the flesh with exchangeable Ca shows that Ca outcompetes group II metals for root uptake and that the uptake pathway discriminated against group II metals relative to ionic radius, with uptake of Ca > Sr > Ba >> Ra. Flesh and stone analysis showed that movement of group II metals to these components of the plant, after root uptake, was strongly related. This supports the hypothesis that Sr, Ba and Ra are being taken up as analogue elements, and follow the same uptake and translocation pathways, with Ca. Comparison with previously reported data from a native passion fruit supports the use of total soil CRs on natural, undisturbed sites. As exchangeable CRs for Ra reach a saturation value it may be possible to make more precise predictions using selective extraction techniques for contaminated or disturbed sites.


Subject(s)
Anacardiaceae/metabolism , Metals, Alkaline Earth/metabolism , Radium/metabolism , Soil Pollutants, Radioactive/metabolism , Barium/metabolism , Calcium/metabolism , Northern Territory , Strontium/metabolism
5.
PLoS One ; 10(5): e0125503, 2015.
Article in English | MEDLINE | ID: mdl-25955826

ABSTRACT

Members of the eukaryotic PIEZO family (the human orthologs are noted hPIEZO1 and hPIEZO2) form cation-selective mechanically-gated channels. We characterized the selectivity of human PIEZO1 (hPIEZO1) for alkali ions: K+, Na+, Cs+ and Li+; organic cations: TMA and TEA, and divalents: Ba2+, Ca2+, Mg2+ and Mn2+. All monovalent ions permeated the channel. At a membrane potential of -100 mV, Cs+, Na+ and K+ had chord conductances in the range of 35-55 pS with the exception of Li+, which had a significantly lower conductance of ~ 23 pS. The divalents decreased the single-channel permeability of K+, presumably because the divalents permeated slowly and occupied the open channel for a significant fraction of the time. In cell-attached mode, 90 mM extracellular divalents had a conductance for inward currents carried by the divalents of: 25 pS for Ba2+ and 15 pS for Ca2+ at -80 mV and 10 pS for Mg2+ at -50 mV. The organic cations, TMA and TEA, permeated slowly and attenuated K+ currents much like the divalents. As expected, the channel K+ conductance increased with K+ concentration saturating at ~ 45 pS and the KD of K+ for the channel was 32 mM. Pure divalent ion currents were of lower amplitude than those with alkali ions and the channel opening rate was lower in the presence of divalents than in the presence of monovalents. Exposing cells to the actin disrupting reagent cytochalasin D increased the frequency of openings in cell-attached patches probably by reducing mechanoprotection.


Subject(s)
Ion Channels/metabolism , Membrane Potentials/physiology , Metals, Alkali/metabolism , Metals, Alkaline Earth/metabolism , Cations, Divalent , Cations, Monovalent , Cell Membrane Permeability/drug effects , Cytochalasin D/pharmacology , Gene Expression , HEK293 Cells , Humans , Ion Channels/genetics , Ion Transport/drug effects , Membrane Potentials/drug effects , Nucleic Acid Synthesis Inhibitors/pharmacology , Patch-Clamp Techniques , Plasmids/chemistry , Plasmids/metabolism , Quaternary Ammonium Compounds/metabolism , Tetraethylammonium/metabolism , Transfection
6.
PLoS One ; 10(5): e0128100, 2015.
Article in English | MEDLINE | ID: mdl-26020945

ABSTRACT

Developing and deploying cotton cultivars with high nutrient uptake, use efficiency and tolerance to nutrient related soil stresses is desirable to assist sustainable soil management. Genetic variation, heritability, selection response and quantitative trait loci (QTLs) were investigated for five macronutrients (P, K, Ca, Mg, S) and five micronutrients (Fe, Mn, B, Zn, and Cu) in a recombinant inbred line (RIL) population from an inter-specific cross between Gossypium hirsutum cv. Guazuncho 2, and G. barbadense accession VH8-4602. Na and K/Na ratio were also studied as the imbalance between Na and other nutrients is detrimental to cotton growth and development. The concentrations of nutrients were measured for different plant parts of the two parents and for leaf samples of the whole population collected at early to peak flowering in field experiments over two years in a sodic Vertosol soil. Parental contrast was large for most nutrient concentrations in leaves when compared with other plant parts. Segregation for leaf nutrient concentration was observed within the population with transgression for P, K, K/Na ratio and all micronutrients. Genotypic difference was the major factor behind within-population variation for most nutrients, while narrow sense heritability was moderate (0.27 for Mn and Cu, and 0.43 for B). At least one significant QTL was identified for each nutrient except K and more than half of those QTLs were clustered on chromosomes 14, 18 and 22. Selection response was predicted to be low for P and all micronutrients except B, high for K, Na and B, and very high for K/Na ratio. Correlations were more common between macronutrients, Na and K/Na ratio where the nature and strength of the relations varied (r=-0.69 to 0.76). We conclude that there is sufficient genetic diversity between these two tetraploid cotton species that could be exploited to improve cotton nutrient status by introgressing species-unique favourable alleles.


Subject(s)
Chromosomes, Plant , Gossypium/genetics , Metals, Alkaline Earth/metabolism , Metals, Heavy/metabolism , Plant Leaves/genetics , Quantitative Trait Loci , Alleles , Cations, Divalent , Cations, Monovalent , Chromosome Mapping , Crosses, Genetic , Genetic Variation , Gossypium/metabolism , Phenotype , Plant Leaves/metabolism
7.
Biometals ; 28(2): 391-400, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25744028

ABSTRACT

Twenty-five days after the disaster at the Fukushima Daiichi nuclear power plant in 2011, we collected samples of the green macroalga Bryopsis maxima from the Pacific coast of Japan. Bryopsis maxima is a unicellular, multinuclear, siphonous green macroalga. Radiation analysis revealed that B. maxima emitted remarkably high gamma radiation of (131)I, (134)Cs, (137)Cs, and (140)Ba as fission products of (235)U. Interestingly, B. maxima contained naturally occurring radionuclides derived from (226)Ra and (228)Ra. Analysis of element content revealed that B. maxima accumulates many ocean elements, especially high quantities of the alkaline earth metals Sr (15.9 g per dry-kg) and Ba (3.79 g per dry-kg), whereas Ca content (12.5 g per dry-kg) was lower than that of Sr and only 61 % of the mean content of 70 Japanese seaweed species. Time-course analysis determined the rate of radioactive (85)Sr incorporation into thalli to be approximately 0.13 g Sr per dry-kg of thallus per day. Subcellular fractionation of B. maxima cells showed that most of the (85)Sr was localized in the soluble fraction, predominantly in the vacuole or cytosol. Given that (85)Sr radioactivity was permeable through a dialysis membrane, the (85)Sr was considered to be a form of inorganic ion and/or bound with a small molecule. Precipitation analysis with sodium sulfate showed that more than 70% of the Sr did not precipitate as SrSO4, indicating that a proportion of the Sr may bind with small molecules in B. maxima.


Subject(s)
Cesium Radioisotopes/metabolism , Chlorophyta/metabolism , Metals, Alkaline Earth/metabolism , Microalgae/metabolism , Cells, Cultured , Fukushima Nuclear Accident , Japan
8.
Chemosphere ; 119: 778-785, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25194478

ABSTRACT

Alkali and alkaline earth metallic (AAEM) species water leaching and Cu(II) sorption by biochar prepared from two invasive plants, Spartina alterniflora (SA) and water hyacinth (WH), were explored in this work. Significant amounts of Na and K can be released (maximum leaching for Na 59.0 mg g(-1) and K 79.9 mg g(-1)) from SA and WH biochar when they are exposed to contact with water. Cu(II) removal by biochar is highly related with pyrolysis temperature and environmental pH with 600-700 °C and pH of 6 showing best performance (29.4 and 28.2 mg g(-1) for SA and WH biochar). Cu(II) sorption exerts negligible influence on Na/K/Mg leaching but clearly promotes the release of Ca. Biochars from these two plant species provide multiple benefits, including nutrient release (K), heavy metal immobilization as well as promoting the aggregation of soil particles (Ca) for soil amelioration. AAEM and Cu(II) equilibrium concentrations in sorption were analyzed by positive matrix factorization (PMF) to examine the factors underlying the leaching and sorption behavior of biochar. The identified factors can provide insightful understanding on experimental phenomena.


Subject(s)
Charcoal/metabolism , Copper/metabolism , Environmental Restoration and Remediation/methods , Metals, Alkali/metabolism , Metals, Alkaline Earth/metabolism , Soil Pollutants/metabolism , Charcoal/chemistry , Eichhornia/chemistry , Introduced Species , Poaceae/chemistry , Principal Component Analysis
9.
Sci Rep ; 3: 1865, 2013.
Article in English | MEDLINE | ID: mdl-23689559

ABSTRACT

Benzoquinones (BQ) have important functions in many biological processes. In alkaline environments, BQs can be hydroxylated at quinoid ring proton positions. Very little is known about the chemical reaction leading to these structural transformations as well as about the properties of the obtained hydroxyl benzoquinones. We analyzed the behavior of the naturally occurring 2,6-dimethoxy-1,4-benzoquinone under alkaline conditions and show that upon substitution of methoxy-groups, poly-hydroxyl-derivatives (OHBQ) are formed. The emerging compounds with one or several hydroxyl-substituents on single or fused quinone-rings exist in oxidized or reduced states and are very stable under physiological conditions. In comparison with the parent BQs, OHBQs are stronger radical scavengers and redox switchable earth-alkaline metal ligands. Considering that hydroxylated quinones appear as biosynthetic intermediates or as products of enzymatic reactions, and that BQs present in food or administered as drugs can be hydroxylated by enzymatic pathways, highlights their potential importance in biological systems.


Subject(s)
Benzoquinones/pharmacology , Calcium/metabolism , Free Radical Scavengers/pharmacology , Hydroxyl Radical/chemistry , Metals, Alkaline Earth/metabolism , Antioxidants/chemistry , Antioxidants/pharmacology , Benzoquinones/chemistry , Chelating Agents/chemistry , Chelating Agents/pharmacology , Electrochemistry , Electron Spin Resonance Spectroscopy , Free Radical Scavengers/chemistry , Hydroxylation , Ligands , Magnetic Resonance Spectroscopy , Oxidants/chemistry , Oxidants/pharmacology , Oxidation-Reduction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
10.
Chemosphere ; 89(9): 1066-76, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22726424

ABSTRACT

We analyzed the ability of freshwater taxa of the genus Ulva (Ulvaceae, Chlorophyta) to serve as bioindicators of metal in lakes and rivers. Changes in heavy metal (Ni, Cd and Pb) and alkaline earth metal (Ca and Mg) concentrations in freshwater Ulva thalli were investigated during the period from June to August 2010. The study was conducted in two ecosystems in Western Poland, the Malta lake (10 sites) and the Nielba river (six sites). Three components were collected for each sample, including water, sediment and Ulva thalli. The average concentrations of metals in the water sample and in the macroalgae decreased in the following order: Ca>Mg>Ni>Pb>Cd. The sediment revealed a slightly altered order: Ca>Mg>Pb>Ni>Cd. Ca and Mg were found at the highest concentrations in thalli due to the presence of carbonate on its surface. Among the examined heavy metals in thalli, Ni was in the highest concentration, and Cd found in the lowest concentration. There were statistically significant correlations between the levels of metals in macroalgae, water and sediment. Freshwater populations of Ulva exhibited a greater efficiency to bioaccumulate nickel as compared to species derived from marine ecosystems.


Subject(s)
Environmental Monitoring/methods , Metals, Alkaline Earth/metabolism , Metals, Heavy/metabolism , Ulva/metabolism , Water Pollutants, Chemical/metabolism , Fresh Water
11.
ACS Chem Biol ; 6(9): 934-42, 2011 Sep 16.
Article in English | MEDLINE | ID: mdl-21736285

ABSTRACT

A majority of enzymes show a high degree of specificity toward a particular metal ion in their catalytic reaction. However, Type II restriction endonuclease (REase) R.KpnI, which is the first member of the HNH superfamily of REases, exhibits extraordinary diversity in metal ion dependent DNA cleavage. Several alkaline earth and transition group metal ions induce high fidelity and promiscuous cleavage or inhibition depending upon their concentration. The metal ions having different ionic radii and co-ordination geometries readily replace each other from the enzyme's active site, revealing its plasticity. Ability of R.KpnI to cleave DNA with both alkaline earth and transition group metal ions having varied ionic radii could imply utilization of different catalytic site(s). However, mutation of the invariant His residue of the HNH motif caused abolition of the enzyme activity with all of the cofactors, indicating that the enzyme follows a single metal ion catalytic mechanism for DNA cleavage. Indispensability of His in nucleophile activation together with broad cofactor tolerance of the enzyme indicates electrostatic stabilization function of metal ions during catalysis. Nevertheless, a second metal ion is recruited at higher concentrations to either induce promiscuity or inhibit the DNA cleavage. Regulation of the endonuclease activity and fidelity by a second metal ion binding is a unique feature of R.KpnI among REases and HNH nucleases. The active site plasticity of R.KpnI opens up avenues for redesigning cofactor specificities and generation of mutants specific to a particular metal ion.


Subject(s)
DNA Cleavage , Deoxyribonucleases, Type II Site-Specific/chemistry , Deoxyribonucleases, Type II Site-Specific/metabolism , Metals, Alkaline Earth/metabolism , Transition Elements/metabolism , Biocatalysis , Catalytic Domain , Ions/chemistry , Ions/metabolism , Metals, Alkaline Earth/chemistry , Substrate Specificity , Transition Elements/chemistry
12.
Plant Physiol Biochem ; 49(5): 494-8, 2011 May.
Article in English | MEDLINE | ID: mdl-21295989

ABSTRACT

In plants, Cd causes perturbation of root metal uptake and is known to interfere with the metal translocation to the shoot. The most significant effect is the strongly reduced transport of Fe. Fe accumulation in roots under Cd stress revealed that it is not the Fe acquisition but the Fe loading to xylem elements that is blocked by Cd, which can be a result of competition between Fe and Cd for the transporters. However, in animal cells as well as in plant stomata guard cells, Cd was shown to move through Ca channels. To clarify whether the perturbation of metal translocation/xylem loading caused by Cd show any regularity, translocation ability was tested by the determination of the metal content in leaves of hydroponically cultured (» Hoagland nutrient solution, Fe source: 10 µM Fe((III))-citrate) poplar plants grown for three weeks with or without 10 µM Cd(NO3)2 treatment. Metals could be classified into two groups according to the behavior of their translocation under Cd treatment: alkaline earth metals (except Mg), Zn and Mn were influenced similarly to Ca, but other transition metals (together with alkali metals and Al) behaved like the Fe. Based on the translocation pattern, Cd seems to inhibit the transport of Ca-like metals competitively, but a different type of inhibition is exerted on the transition metal transport, with which Cd can share a common translocation system. The strongly decreased translocation of chelator-dependent transition metals may indicate Cd related disturbances in signalling pathways and gene expression of xylem transporters or chelators.


Subject(s)
Cadmium/metabolism , Iron/metabolism , Metals, Alkaline Earth/metabolism , Plant Leaves/metabolism , Populus/metabolism , Analysis of Variance , Biological Transport , Cadmium/analysis , Citric Acid/metabolism , Iron/analysis , Manganese/analysis , Manganese/metabolism , Metals, Alkali/analysis , Metals, Alkali/metabolism , Metals, Alkaline Earth/analysis , Plant Leaves/chemistry , Plant Leaves/growth & development , Populus/chemistry , Populus/growth & development , Water/chemistry , Xylem/metabolism
13.
Biochem J ; 435(2): 391-9, 2011 Apr 15.
Article in English | MEDLINE | ID: mdl-21265816

ABSTRACT

CASQ (calsequestrin) is a Ca2+-buffering protein localized in the muscle SR (sarcoplasmic reticulum); however, it is unknown whether Ca2+ binding to CASQ2 is due to its location inside the SR rich in Ca2+ or due to its preference for Ca2+ over other ions. Therefore a major aim of the present study was to determine how CASQ2 selects Ca2+ over other metal ions by studying monomer folding and subsequent aggregation upon exposure to alkali (monovalent), alkaline earth (divalent) and transition (polyvalent) metals. We additionally investigated how CPVT (catecholaminergic polymorphic ventricular tachycardia) mutations affect CASQ2 structure and its molecular behaviour when exposed to different metal ions. Our results show that alkali and alkaline earth metals can initiate similar molecular compaction (folding), but only Ca2+ can promote CASQ2 to aggregate, suggesting that CASQ2 has a preferential binding to Ca2+ over all other metals. We additionally found that transition metals (having higher co-ordinated bonding ability than Ca2+) can also initiate folding and promote aggregation of CASQ2. These studies led us to suggest that folding and formation of higher-order structures depends on cationic properties such as co-ordinate bonding ability and ionic radius. Among the CPVT mutants studied, the L167H mutation disrupts the Ca2+-dependent folding and, when folding is achieved by Mn2+, L167H can undergo aggregation in a Ca2+-dependent manner. Interestingly, domain III mutants (D307H and P308L) lost their selectivity to Ca2+ and could be aggregated in the presence of Mg2+. In conclusion, these studies suggest that CPVT mutations modify CASQ2 behaviour, including folding, aggregation/polymerization and selectivity towards Ca2+.


Subject(s)
Calsequestrin/metabolism , Cations/metabolism , Mutant Proteins/metabolism , Myocardium/metabolism , Tachycardia, Ventricular/genetics , Amino Acid Sequence , Calcium/metabolism , Calcium/pharmacology , Calsequestrin/chemistry , Calsequestrin/genetics , Calsequestrin/physiology , Humans , Metals, Alkaline Earth/metabolism , Metals, Alkaline Earth/pharmacology , Models, Molecular , Molecular Probe Techniques , Molecular Sequence Data , Mutant Proteins/analysis , Mutation, Missense/physiology , Protein Conformation/drug effects , Protein Folding , Protein Multimerization/genetics , Protein Structure, Tertiary/genetics , Protein Structure, Tertiary/physiology , Substrate Specificity , Tachycardia, Ventricular/metabolism
14.
J Biol Inorg Chem ; 16(4): 523-37, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21197556

ABSTRACT

Anticoagulation factor II (ACF II) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X (FXa)-binding protein with both anticoagulant and hypotensive activities. The thermodynamics of the binding of alkaline earth metal ions to ACF II and their effects on the stability of ACF II and the binding of ACF II to FXa were investigated by isothermal titration calorimetry, fluorescence, differential scanning calorimetry, and surface plasmon resonance. The binding of ACF II to FXa does not have an absolute requirement for Ca(2+). Mg(2+), Sr(2+), and Ba(2+) can induce the binding of ACF II to FXa. The radii of the cations bound in ACF II crucially affect the binding affinity of ACF II for cations and the structural stability of ACF II against guanidine hydrochloride and thermal denaturation, whereas the radii of cations bound in FXa markedly affect the binding affinity between ACF II and FXa. The binding affinities of ACF II for cations and the capacities of metal-induced stabilization of ACF II follow the same trend: Ca(2+) > Sr(2+) > Ba(2+). The metal-induced binding affinities of ACF II for FXa follow the trend Mg(2+) > Ca(2+) > Sr(2+) > Ba(2+). Although Mg(2+) shows significantly low binding affinity with ACF II, Mg(2+) is the most effective to induce the binding of ACF II with FXa. Our observations suggest that in blood the bindings of Ca(2+) in two sites of ACF II increase the structural stability of ACF II, but these bindings are not essential for the binding of ACF II with FXa, and that the binding of Mg(2+) and Ca(2+) to FXa may be essential for the recognition between FXa and ACF II. Like Ca(2+), the abundant Mg(2+) in blood also plays an important role in the anticoagulation of ACF II.


Subject(s)
Crotalid Venoms/chemistry , Crotalid Venoms/metabolism , Factor X/metabolism , Metals, Alkaline Earth/metabolism , Venoms/chemistry , Binding Sites , Crotalid Venoms/antagonists & inhibitors , Factor X/chemistry , Guanidine/antagonists & inhibitors , Guanidine/pharmacology , Ions/metabolism , Models, Molecular , Protein Conformation , Protein Denaturation/drug effects , Structure-Activity Relationship , Thermodynamics
15.
Br J Pharmacol ; 163(8): 1605-25, 2011 Aug.
Article in English | MEDLINE | ID: mdl-20718728

ABSTRACT

The design and engineering of organic fluorescent Ca(2+) indicators approximately 30 years ago opened the door for imaging cellular Ca(2+) signals with a high degree of temporal and spatial resolution. Over this time, Ca(2+) imaging has revolutionized our approaches for tissue-level spatiotemporal analysis of functional organization and has matured into a powerful tool for in situ imaging of cellular activity in the living animal. In vivo Ca(2+) imaging with temporal resolution at the millisecond range and spatial resolution at micrometer range has been achieved through novel designs of Ca(2+) sensors, development of modern microscopes and powerful imaging techniques such as two-photon microscopy. Imaging Ca(2+) signals in ensembles of cells within tissue in 3D allows for analysis of integrated cellular function, which, in the case of the brain, enables recording activity patterns in local circuits. The recent development of miniaturized compact, fibre-optic-based, mechanically flexible microendoscopes capable of two-photon microscopy opens the door for imaging activity in awake, behaving animals. This development is poised to open a new chapter in physiological experiments and for pharmacological approaches in the development of novel therapies.


Subject(s)
Brain/physiology , Calcium/analysis , Diagnostic Imaging/methods , Metals, Alkaline Earth/analysis , Microscopy/methods , Photons , Animals , Brain/metabolism , Calcium/metabolism , Calcium/physiology , Humans , Metals, Alkaline Earth/metabolism , Metals, Alkaline Earth/pharmacology , Models, Animal
16.
Biochem Biophys Res Commun ; 393(3): 471-5, 2010 Mar 12.
Article in English | MEDLINE | ID: mdl-20152801

ABSTRACT

Iron deficiency is the most prevalent micronutrient deficiency worldwide. Whereas dietary calcium is known to reduce the bioavailability of iron, the molecular basis of this interaction is not understood. We tested the hypothesis that divalent metal-ion transporter-1 (DMT1)-the principal or only mechanism by which nonheme iron is taken up at the intestinal brush border-is shared also by calcium. We expressed human DMT1 in RNA-injected Xenopus oocytes and examined its activity using radiotracer assays and the voltage clamp. DMT1 did not mediate 45Ca2+ uptake. Instead, we found that Ca2+ blocked the Fe2+-evoked currents and inhibited 55Fe2+ uptake in a noncompetitive manner (K(i) approximately 20 mM). The mechanism of inhibition was independent of voltage and did not involve intracellular Ca2+ signaling. The alkaline-earth metal ions Ba2+, Sr2+, and Mg2+ also inhibited DMT1-mediated iron-transport activity. We conclude that Ca2+ is a low-affinity noncompetitive inhibitor--but not a transported substrate--of DMT1, explaining in part the effect of high dietary calcium on iron bioavailability.


Subject(s)
Calcium/metabolism , Cation Transport Proteins/antagonists & inhibitors , Cation Transport Proteins/metabolism , Animals , Calcium, Dietary/metabolism , Cation Transport Proteins/genetics , Humans , Iron/metabolism , Metals, Alkaline Earth/metabolism , Oocytes , Substrate Specificity , Xenopus
17.
Biochemistry ; 49(6): 1268-80, 2010 Feb 16.
Article in English | MEDLINE | ID: mdl-20052994

ABSTRACT

To investigate the role of the active site copper in Escherichia coli copper amine oxidase (ECAO), we initiated a metal-substitution study. Copper reconstitution of ECAO (Cu-ECAO) restored only approximately 12% wild-type activity as measured by k(cat(amine)). Treatment with EDTA, to remove exogenous divalent metals, increased Cu-ECAO activity but reduced the activity of wild-type ECAO. Subsequent addition of calcium restored wild-type ECAO and further enhanced Cu-ECAO activities. Cobalt-reconstituted ECAO (Co-ECAO) showed lower but significant activity. These initial results are consistent with a direct electron transfer from TPQ to oxygen stabilized by the metal. If a Cu(I)-TPQ semiquinone mechanism operates, then an alternative outer-sphere electron transfer must also exist to account for the catalytic activity of Co-ECAO. The positive effect of calcium on ECAO activity led us to investigate the peripheral calcium binding sites of ECAO. Crystallographic analysis of wild-type ECAO structures, determined in the presence and absence of EDTA, confirmed that calcium is the normal ligand of these peripheral sites. The more solvent exposed calcium can be easily displaced by mono- and divalent cations with no effect on activity, whereas removal of the more buried calcium ion with EDTA resulted in a 60-90% reduction in ECAO activity and the presence of a lag phase, which could be overcome under oxygen saturation or by reoccupying the buried site with various divalent cations. Our studies indicate that binding of metal ions in the peripheral sites, while not essential, is important for maximal enzymatic activity in the mature enzyme.


Subject(s)
Amine Oxidase (Copper-Containing)/chemistry , Escherichia coli Proteins/chemistry , Metals, Heavy/chemistry , Amine Oxidase (Copper-Containing)/antagonists & inhibitors , Amine Oxidase (Copper-Containing)/metabolism , Amino Acid Sequence , Calcium/chemistry , Calcium/metabolism , Catalytic Domain/genetics , Copper/chemistry , Crystallography, X-Ray , Dihydroxyphenylalanine/analogs & derivatives , Dihydroxyphenylalanine/chemistry , Edetic Acid/chemistry , Enzyme Activation/genetics , Escherichia coli Proteins/antagonists & inhibitors , Escherichia coli Proteins/metabolism , Humans , Metals, Alkaline Earth/chemistry , Metals, Alkaline Earth/metabolism , Metals, Heavy/metabolism , Molecular Sequence Data , Oxidation-Reduction , Protein Binding/genetics
18.
J Biol Inorg Chem ; 14(4): 559-71, 2009 May.
Article in English | MEDLINE | ID: mdl-19184130

ABSTRACT

Anticoagulation factor I (ACF I) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X (FXa)-binding protein that binds in a Ca(2+)-dependent fashion with marked anticoagulant activity. The thermodynamics of the binding of alkaline earth metal ions to ACF I and the effects of alkaline earth metal ions on the guanidine hydrochloride (GdnHCl)-induced unfolding of ACF I and the binding of ACF I to FXa were studied by isothermal titration calorimetry, fluorescence, circular dichroism, and surface plasmon resonance, respectively. The results indicate that the ionic radii of the cations occupying Ca(2+)-binding sites in ACF I crucially affect the binding affinity of ACF I for alkaline earth metal ions as well as the structural stability of ACF I against GdnHCl denaturation. Sr(2+) and Ba(2+), with ionic radii larger than the ionic radius of Ca(2+), can bind to Ca(2+)-free ACF I (apo-ACF I), while Mg(2+), with an ionic radius smaller than that of Ca(2+), shows significantly low affinity for the binding to apo-ACF I. All bindings of Ca(2+), Sr(2+), and Ba(2+) ions in two sites of ACF I are mainly enthalpy-driven and the entropy is unfavorable for them. Sr(2+)-stabilized ACF I exhibits slightly lower resistance to GdnHCl denaturation than Ca(2+)-ACF I, while Ba(2+)-stabilized ACF I exhibits much lower resistance to GdnHCl denaturation than Ca(2+)-ACF I. Mg(2+) and Sr(2+), with ionic radii close to that of Ca(2+), can bind to FXa and therefore also induce the binding of ACF I to FXa, whereas Ba(2+), with a much larger ionic radius than Ca(2+), cannot support the binding of ACF I with FXa. Our observations suggest that bindings of Ca(2+), Sr(2+), and Ba(2+) ions in two sites of ACF I increase the structural stability of ACF I, but these bindings are not essential for the binding of ACF I with FXa, and that the binding of Mg(2+), Ca(2+), and Sr(2+) ions to FXa may be essential for the recognition between FXa and ACF I.


Subject(s)
Agkistrodon , Crotalid Venoms/chemistry , Crotalid Venoms/metabolism , Factor Xa/metabolism , Metals, Alkaline Earth/metabolism , Protein Structure, Tertiary , Animals , Calcium/metabolism , Cattle , Factor Xa/chemistry , Guanidine/chemistry , Ions/chemistry , Ions/metabolism , Metals, Alkaline Earth/chemistry , Models, Molecular , Protein Binding , Protein Denaturation , Protein Folding , Thermodynamics
19.
J Trace Elem Med Biol ; 22(3): 234-41, 2008.
Article in English | MEDLINE | ID: mdl-18755399

ABSTRACT

The main objective of this work was to evaluate the interaction between selenium concentration in both commercial and Se-enriched eggs and other essential/toxic elements (Ca, Mg, Fe, Zn, Pb, and Cd), taking into account a possible synergic action of iodine. Commercial eggs were purchased from several sale points or directly from the producers (farmyard eggs). Fortified eggs were obtained by supplementing chickenfeed for 6 weeks with Se as sodium selenite (1.0 microg/g Se) or Se plus iodine (1.0 microg/g Se+3.7 microg/g I). Se in experimental egg yolks significantly increased over the basic value by 39% in the Se group and 61% in the Se+I group, suggesting that I addition may enhance Se absorption. Levels of Se in commercial yolks were identical in free-range, barn or battery eggs, but significantly lower in farmyard and higher in organic eggs where the Se content approximated that found in Se fortified eggs. A significant reduction in Cd was observed in Se+I treated yolks compared to both control and Se alone diet, thus suggesting a high sensitivity of Cd to the detoxifying effect of Se combined with I. Furthermore, Se+I supplementation was associated with a significant Zn reduction, a finding which needs clarification to avoid attempts to maximize one component affecting the levels of other essential elements.


Subject(s)
Chickens/metabolism , Egg Yolk/chemistry , Egg Yolk/metabolism , Eggs/analysis , Food, Fortified/analysis , Metals, Alkaline Earth/chemistry , Metals, Heavy/chemistry , Selenium/chemistry , Selenium/metabolism , Animals , Female , Metals, Alkaline Earth/metabolism , Metals, Heavy/metabolism
20.
Luminescence ; 22(6): 575-80, 2007.
Article in English | MEDLINE | ID: mdl-17768715

ABSTRACT

In the present work, the generation mechanism of reactive oxygen species (ROS) on calcium peroxide (CaO(2)) was studied. A very intense chemiluminescence (CL) signal was observed when adding an aqueous solution of luminol or 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo[1,2alpha]-pyrazin-3-one hydrochloride (MCLA) to a suspension of CaO(2). The ROS released on CaO(2) were thought to be oxidizing agents leading to CL, and were characterized by CL, UV-visible (UV-vis) spectra and the effective scavengers of the special ROS. From experimental results, the hydroxyl (.OH) and superoxide (.O(2) (-)) radicals were suggested to exist on the surface of CaO(2). A reaction scheme for the formation of the ROS on CaO(2) was also proposed and discussed. Of more interest was the finding that the CaO(2) which released the .OH and .O(2) (-) on the surface exhibited good transition properties compared with alkaline-earth metal peroxides of the same group (MgO(2), BaO(2)).


Subject(s)
Luminescence , Luminescent Measurements/methods , Peroxides/metabolism , Reactive Oxygen Species/metabolism , Spectrophotometry, Ultraviolet/methods , Free Radical Scavengers/chemistry , Free Radical Scavengers/metabolism , Hydroxyl Radical/chemistry , Hydroxyl Radical/metabolism , Imidazoles/chemistry , Luminol/chemistry , Metals, Alkaline Earth/chemistry , Metals, Alkaline Earth/metabolism , Oxygen/chemistry , Oxygen/metabolism , Peroxides/chemistry , Pyrazines/chemistry , Reactive Oxygen Species/chemistry , Solutions/chemistry , Time Factors , Water/chemistry
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