Comparative insight into the Zn(II)-, Cd(II)- and Cu(I)-binding features of the protozoan Tetrahymena pyriformis MT1 metallothionein

Tetrahymena pyriformis MT1 (TpyMT1) is a model among ciliate metallothioneins (MTs). Here, we report on the analytic (ICP-AES, GC-FPD), spectroscopic (CD, UV-Vis, Raman) and spectrometric (ESI-MS) characterization of its recombinant Cd(II)-, Zn(II)- and Cu(I)-complexes, and of those formed during in vitro Zn/Cd and Zn/Cu replacement. In the presence of Cd(II), TpyMT1 renders a major Cd 11-TpyMT1 species, which is also the final step reached in the in vitro Zn/Cd exchange process in Zn 11-TpyMT1. Spectroscopic data supports a different folding of the isostoichiometric Cd 11- and Zn 11-TpyMT1 complexes. Unexpectedly, TpyMT1 biosynthesis in Zn(II)-rich cultures was sensitive to the aeration degree, so that high oxygenation rendered undermetalated, partially-oxidized, complexes (Zn9-TpyMT1). Biosynthesis in Cu(I)-rich media rendered extremely heterogeneous mixtures of CuxZny-species (x+y=8-20), where the higher the aeration, the higher the Zn(II) content. The complexity of these samples was reproduced during the Zn/Cu replacement, as the number of generated species increased gradually with the addition of copper to Zn(11)-TpyMT1. According to our results, a clear preference of TpyMT1 for Cd(II) binding, rather than for Zn(II), and especially Cu(I) can be postulated. This character is totally consistent with the induction pattern of the TpyMT1 gene and the postulated role of TpyMT1 in Cd-detoxification.     

Antimicrobial activity of ZnII, CdII, and HgII complexes of 1,2-bis-[2-(5-R)-1H-benzimidazolyl]-1,2-ethanediols and 1,4-bis-[2-(5-R)-1H-benzimidazolyl]-1,2,3,4-butanetetraols

1,2-Bis-[2-(5-H/Me/Cl/NO2)-1H-benzimidazolyl]-1,2-ethanediols (L1-L4), 1,4-bis-[2-(5-H/Me/Cl)-1H-benzimidazolyl]-1,2,3,4-butanetetraols (L5-L7) and their complexes with ZnCl2, CdCl2 and HgCl2 were synthesized and antibacterial activity of the compounds was tested toward Staphylococcus aureus, S. epidermidis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Shigella flexneri, Proteus mirabilis and antifungal activity against Candida albicans. HgII complexes have a considerably higher antimicrobial activity against all microorganisms. Some HgII complexes show higher antifungal activity than clotrimazole toward C. albicans. Zn2(L3)Cl4, Zn2(L4)Cl4, and Cd(L3)Cl2 were moderately effective against S. aureus and S. epidermidis; Cd(L4)Cl2 exhibited a weak activity only against S. epidermidis.     

Zinc and cadmium uptake by the hyperaccumulator Thlaspi caerulescens in contaminated soils and its effects on the concentration and chemical speciation of metals in soil solution

The hyperaccumulator Thlaspi caerulescens J & C Presl. was grown in seven different soils collected from around Europe that had been contaminated with heavy metals by industrial activity or the disposal of sewage sludge to land. Zinc accumulation factors (shoot concentration/initial soil solution concentration) ranged from 3500–85 000 with a mean value of around 36 000. This compares with mean accumulation factors of 636, 66 and 122 for Cd, Ca and Mg, respectively. The concentration of Zn in the shoots was much greater than in the roots. The total removal of Zn and Cd ranged from 8 to 30 and from 0.02 to 0.5 mg kg^-1 soil, respectively. The Zn concentration in shoots of T. caerulescens correlated, using a curvilinear relationship, with the initial Zn concentration in soil solution (R² = total Zn 0.78; Zn²+ 0.80). There was no relationship between the uptake of Zn and the total Zn concentration in the soil. In most soils, solution pH increased only slightly after growth of T. caerulescens, indicating that acidification was not the mechanism used to mobilise Zn in the soil. Dissolved organic carbon concentrations generally increased but characterisation of the component organic compounds was not attempted. The concentrations of Zn and Cd in soil solution decreased considerably after growth of T. caerulescens. The percentages of Zn and Cd in soil solution present as free ions also decreased. However, the decrease of Zn in soil solution after growth accounted for only about 1% of the total Zn uptake by T. caerulescens. This was much lower than for Cd, Ca and Mg. The results suggest that either T. caerulescens was highly efficient at mobilising Zn which was not soluble initially, or the soils used had large buffering capacities to replenish soil solution Zn within a short time. This work highlights the need to investigate the role of root exudates on the mobilisation of Zn and Cd in soils by the hyperaccumulator T. caerulescens.     

Accumulation of cadmium and selected elements in flax seed grown on a calcareous soil

Seed of flax (Linum usitatissimum L.) grown on calcareous and neutral soils sometimes accumulates relatively high concentrations of Cd. The influence of a post-flowering application of NH₄NO₃ (115 mg N kg^-1), CdSO₄ (1 mg Cd kg^-1), FeEDDHA (2 mg Fe kg^-1), NaH₂PO₄ (120 mg P kg^-1) and ZnSO₄ (8 mg Zn kg^-1) on seed accumulation of Cd, Fe, N, Mn, P and Zn by flax grown on a Calciaquoll was studied in two experiments under greenhouse conditions. Seed yields were increased by the N and Zn treatments, and the N×Zn interaction was positive. Zinc deficiency delayed flowering and boll formation by up to 20 days and reduced seed size. In the absence of added Cd, seed accumulated up to 0.33 mg Cd kg^-1. This Cd accumulation was reduced by approximately 50 and 17% by added Zn and Fe, respectively, but was little affected by P fertilizer and post-flowering N stress. In the presence of added Cd, seed Cd exceeded 3.3 mg Cd kg^-1, and the antagonistic effects of Fe and Zn on seed Cd were absent. Seed N, P, Fe and Zn concentrations were increased on average by 10, 45, 31 and 97% by the N, P, Fe and Zn fertilizer treatments, respectively. FeEDDHA reduced seed Mn concentration by approximately 58%. However, seed Mn concentration was much less than that found in vegetative tissue at flowering. Soil-applied Zn may reduce seed Cd concentration in flax under field conditions, and may increase marketability of flax for food use.     

Cd、Zn及其交互作用对互花米草中重金属的积累、亚细胞分布及化学形态的影响

通过盆栽试验,研究了Cd、Zn及其交互作用下互花米草中Cd、Zn的含量及积累量,并分析了Cd、Zn在互花米草中的亚细胞分布及化学形态。结果表明：Cd-Zn处理组互花米草地上部及根部Cd含量显著高于Cd处理组;Cd-Zn处理组根部Zn含量显著低于Zn处理组,但地上部差异不显著,说明Zn促进Cd的吸收,Cd抑制Zn的吸收。Cd-Zn处理组互花米草地上部Cd积累量显著高于Cd处理组,但是根部Cd积累量却显著低于Cd处理组;Zn处理组地上部及根部Zn积累量均显著高于对照组及Cd-Zn处理组。Cd单因素胁迫下,Cd主要分布在细胞壁,Cd-Zn交互作用下,Cd在胞液中的分配比例高于其他细胞组分;Zn单因素及Cd-Zn交互作用下,Zn在胞液中的分配比例均较高,总的分配比例呈现以下趋势：胞液>细胞壁>细胞器,说明Zn的添加影响了Cd的亚细胞分布,Cd的出现对Zn在互花米草细胞中的分布影响不明显。Cd和Zn在互花米草叶中主要以氯化钠提取态存在,表明互花米草中Cd和Zn多以果胶酸盐结合态或蛋白质结合态存在。     

Regulation of phytochelatin synthesis by zinc and cadmium in marine green alga,Dunaliella tertiolecta

Although Cd(2+) is a more effective inducer of phytochelatin (PC) synthesis than Zn(2+) in higher plants, we have observed greater induction of PC synthesis by Zn(2+) than Cd(2+) in the marine green alga, Dunaliella tertiolecta. To elucidate this unique regulation of PC synthesis by Zn(2+), we investigated the effects of Zn(2+) and Cd(2+) on the activities of both phytochelatin synthase (PC synthase) and enzymes in the GSH biosynthetic pathway. PC synthase was more strongly activated by Cd(2+) than by Zn(2+), but the difference was not very big. On the other hand, gamma-glutamylcysteine synthetase (gamma-ECS) and glutathione synthetase (GS) were activated by both heavy metals, but their activities were higher in Zn-treated cells than in Cd-treated cells. Dose-dependent stimulation of intracellular reactive oxygen species (ROS) production was observed with Zn(2+), but not Cd(2+) treatment. These results suggest that Zn(2+) strongly promotes the synthesis of GSH through indirect activation of gamma-ECS and GS by stimulating ROS generation. This acceleration of the flux rate for GSH synthesis might mainly contribute to high level PC synthesis.     

Cadmium uptake, translocation and tolerance in the hyperaccumulator Arabidopsis halleri

Arabidopsis halleri is a well-known zinc (Zn) hyperaccumulator, but its status as a cadmium (Cd) hyperaccumulator is less certain. Here, we investigated whether A. halleri can hyperaccumulate Cd and whether Cd is transported via the Zn pathway. Growth and Cd and Zn uptake were determined in hydroponic experiments with different Cd and Zn concentrations. Short-term uptake and root-to-shoot transport were measured with radioactive 109Cd and 65Zn labelling. A. halleri accumulated > 1000 mg Cd kg(-1) in shoot dry weight at external Cd concentrations >or= 5 microm, but the short-term uptake rate of 109Cd was much lower than that of 65Zn. Zinc inhibited short-term 109Cd uptake kinetics and root-to-shoot translocation, as well as long-term Cd accumulation in shoots. Uptake of 109Cd and 65Zn were up-regulated, respectively, by low iron (Fe) or Zn status. A. halleri was much less tolerant to Cd than to Zn. We conclude that A. halleri is able to hyperaccumulate Cd partly, at least, through the Zn pathway, but the mechanisms responsible for cellular Zn tolerance cannot detoxify Cd effectively.     

The influence of zinc on apical uptake of cadmium in the gills and cadmium influx to the circulatory system in zebrafish (Danio rerio)

Zn (0-16 microM) effects on apical Cd uptake from the water into the branchial epithelium and influx of Cd from the water to the circulatory system in zebrafish (Danio rerio) were studied in three experiments. Apical Cd uptake was decreased by Zn in all three experiments. In fish exposed to 1-600 nM Cd (experiment 1), apical Cd uptake showed saturation kinetics at 2 and 4 microM Zn, and a competitive interaction was indicated. At 16 microMZn, Cd uptake increased linearly. Cadmium influx did not show saturation kinetics, but was inhibited by 16 microM Zn at low Cd exposure concentrations. In fish exposed to 0.1-600 nM Cd (experiment 2), Cd uptake was inhibited by 16 microM Zn, whereas at 30 nM Cd uptake was inhibited by 2 microM Zn. Similarly, 2 microM Zn did not influence Cd uptake in fish exposed to 0.1-2 nM Cd (experiment 3), whereas 2 microM Zn inhibited uptake at 8-30 nM Cd. Zinc also inhibited Cd influx at higher Cd concentrations. However, at lower Cd exposures, a Zn-induced increased influx was indicated. Zinc influences the Cd uptake and influx processes at several sites in the branchial epithelial cells, indicating that influx of Zn2+ and Cd2+ occurs through common pathways.     

Selective, quantitative detection of cadmium and/or zinc by use of BTAN (2-aminobenzothiazolylazo-beta-naphthol)

BTAN (Sumi Y et al. (1982) Histochemistry 73:481) was investigated as a histochemical Cd/Zn chelator. Cd-BTAN exhibits a main peak about 635 nm, while Zn-BTAN exhibits a main peak about 644 nm. The isobestic wavelength for Cd-BTAN and Zn-BTAN is 638 nm. The microscopical detection limit for Cd is about 25 amol/microns 2, and for Zn about 5 amol/microns 2. The relation between metal and bound chelator is fairly linear at a BTAN concentration more than 10-fold the metal concentration. Histochemical localization was fair to good, with a crystal size of up to 0.2-0.3 micron. The chelate was unaffected by hydrophilic and largely also by hydrophobic mounting media. The original staining procedure proved erratic and was modified. Posttreatment with oxine to selectively demonstrate Cd in the presence of Zn (Sumi Y et al. 1982) seriously reduced the staining intensity. Post-treatment for 8-15 min with HCl, 0.5 mol/l, in 50% ethanol removed Cd-BTAN completely with little reduction of Zn staining intensity, even from sites with 5x as much Cd as Zn. It is concluded that BTAN permits direct quantitative detection of (Zn + Cd). Provided certain precautions are taken quantitative detection of Zn and quantitation of Cd in mixed Zn/Cd sites is possible by microphotometry of the stained section before and after differentiation for 8-15 min with the HCl/50% ethanol medium.     

Characteristics of cadmium uptake in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens

Uptake of Cd and Zn by intact seedlings of two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens was characterized using radioactive tracers. Uptake of Cd and Zn at 2 degrees C was assumed to represent mainly apoplastic binding in the roots, whereas the difference in uptake between 22 degrees C and 2 degrees C represented metabolically dependent influx. There was no significant difference between the two ecotypes in the apoplastic binding of Cd or Zn. Metabolically dependent uptake of Cd was 4.5-fold higher in the high Cd-accumulating ecotype, Ganges, than in the low Cd-accumulating ecotype, Prayon. By contrast, there was only a 1.5-fold difference in the Zn uptake between the two ecotypes. For the Ganges ecotype, Cd uptake could be described by Michaelis-Menten kinetics with a V(max) of 143 nmol g(-1) root FW h(-1) and a K(m) of 0.45 microM. Uptake of Cd by the Ganges ecotype was not inhibited by La, Zn, Cu, Co, Mn, Ni or Fe(II), and neither by increasing the Ca concentration. By contrast, addition of La, Zn or Mn, or increasing the Ca concentration in the uptake solution decreased Cd uptake by Prayon. Uptake of Ca was larger in Prayon than in Ganges. The results suggest that Cd uptake by the low Cd-accumulating ecotype (Prayon) may be mediated partly via Ca channels or transporters for Zn and Mn. By contrast, there may exist a highly selective Cd transport system in the root cell membranes of the high Cd-accumulating ecotype (Ganges) of T. caerulescens.     

Concentrations of cadmium,zinc, copper,iron, and metallothionein in liver and kidney of nonhuman primates

To evaluate the species specificity of Cd accumulation and the relationship of Cd with other essential metals and metallothionein (MT), the concentrations of Cd, Zn, Cu, and Fe in the liver and kidney and the MT concentrations in the soluble fractions of the liver and kidney were determined in Cd-uncontaminated nonhuman primates (11 species, 26 individuals) kept in a zoo and two wild-caught Japanese macaques. The compositions of metal-binding proteins in the soluble fractions were also investigated by high-performance liquid chromatography (HPLC). The hepatic Cd concentration was 0.03–14.0 μg/g and the renal Cd concentration was 0.35–99.0 μg/g, both varying greatly and being higher in nonhuman primates, which were more closely related to man. The hepatic Zn concentration was 24.0–176 μg/g and the renal Zn concentration was 13.5–138 μg/g, showing 7- to 10-fold differences, and a correlation (r=0.558, p<0.01) was found between renal Zn and renal Cd concentrations. It was proved that in the liver, MT is more closely correlated with Zn (r=0.795, p<0.001) than with Cd (r=0.492, p<0.01) and that in the kidney MT is correlated with both Cd (r=0.784, p<0.001) and Zn (r=0.742, p<0.001). HPLC analysis of metals bound to MT-like protein in chimpanzees, de Brazza’s monkeys, and Bolivian squirrel monkeys showed that more than 90% of Cd in both the liver and kidney, approx 40% of Zn in liver and 28–69% of Zn in kidney were bound to MT-like protein. The higher percentage Zn was bound to high-molecular protein.     

锌对镉胁迫下马蔺生长、镉积累及生理抗性的影响

通过溶液培养研究了不同浓度锌(Zn)对镉(Cd)胁迫下马蔺(Iris lacteavar. chinen-sis)生长、Cd积累及抗氧化酶活性等的影响.结果表明:加入1~100mg.L-1Zn后,Cd胁迫下马蔺地上部Cd含量变化不大,而地下部Cd含量显著增加并呈先增后降的趋势;1mg.L-1Zn处理下马蔺地下部Cd含量最高,比对照显著增加51.4%;Zn浓度高于1mg.L-1后Cd含量均出现不同程度的下降,但仍高于对照.与单独Cd处理(10mg.L-1)相比,添加低浓度Zn(1~10mg.L-1)后,马蔺地上部生物量和叶绿素含量呈增加趋势,丙二醛(MDA)含量降低;在10mg.L-1Zn处理下,叶绿素a、b含量达到峰值,分别增加5.21%和22.27%,MDA含量降低25.46%,表明低浓度Zn缓解了Cd对马蔺的毒害.随Zn浓度的增加,Zn对Cd毒害的缓解作用逐渐降低,当溶液中Zn达到一定浓度(100mg.L-1)时,马蔺毒害加重,其生物量、叶绿素含量均下降,MDA含量显著增加.在试验胁迫浓度范围内超氧化物歧化酶(SOD)、过氧化物酶(POD)活性始终呈增加趋势,表明马蔺在受到Cd胁迫及Cd-Zn共存的条件下SOD和POD起着重要的抗氧化保护作用.     

重金属离子对凡纳滨对虾肝胰脏、鳃丝和血液SOD活力的影响

研究了3种重金属离子(Cu²⁺、Zn²⁺、Cd²⁺)在96 h内对凡纳滨对虾(Litopenaeus vannamei)对肝胰脏、鳃丝和血液超氧化物歧化酶(SOD)活力的影响.结果表明,凡纳滨对虾SOD活力在3种重金属离子作用下随取样时间变化显著(P＜0.0),Cu²⁺在实验浓度范围内(0.1～1 mg·L^-1),肝胰脏、鳃丝和血液的SOD活力随时间延长呈一峰值变化,Zn²⁺在10 mg·L^-1时对肝胰脏表现为显著抑制作用,Cd²⁺在0. mg·L^-1时对肝胰脏和鳃丝起显著抑制作用,0.2 mg·L^-1对鳃丝SOD活力无显著变化(P＞0.0),其他浓度Zn²⁺(＜10 mg·L^-1)、Cd²⁺(＜0.2 mg·L^-1)对各组织器官SOD活力的影响随时间延长均呈现先升高后下降的趋势.3种重金属离子对凡纳滨对虾肝胰脏、鳃丝、血液SOD活力的影响呈现明显的剂量-时间效应关系.其SOD活力大小顺序为肝胰脏＞鳃丝＞血液,3种重金属离子对凡纳滨对虾伤害大小顺序为Cd²⁺＞Cu²⁺＞Zn²⁺.     

Reversal of Cadmium-Induced Thyroid Dysfunction by Selenium, Zinc, or Their Combination in Rat

The aim of this study was to evaluate the potential benefit of combined treatment with zinc (Zn) and selenium (Se) in reversing cadmium (Cd)-induced thyroid dysfunction compared to Se or Zn treatment alone in rats exposed to Cd. For this purpose, 30 adult male Wistar albino rats were equally divided into control and four treated groups receiving either 200 ppm Cd (as CdCl2), 200 ppm Cd + 500 ppm Zn (as ZnCl2), 200 ppm Cd + 0.1 ppm Se (as Na2SeO3), or 200 ppm Cd + 500 ppm Zn + 0.1 ppm Se in their drinking water for 35 days. The results showed that Cd exposure increased significantly the relative thyroid weight (RTW), the thyroid Cd concentration, and the serum thyroid stimulating hormone (TSH) level, whereas the serum thyroxine (T4) level was decreased compared to control rats. The treatment of Cd-exposed rats with Se alone only partially protected from the Cd-induced decrease in serum T4 level. The treatment of Cd-exposed animals with Zn alone partially protected against Cd-induced thyroid dysfunction by maintaining normal RTW and by decreasing Cd concentration in the thyroid. It also partially prevents Cd-induced decrease in serum T4 level. The combined treatment of Cd-exposed animals with Se and Zn induced a more significant decrease in the thyroid Cd concentration than the Zn supplement and a total correction of the RTW. This treatment was also more effective than that with Se or Zn alone in reversing Cd-induced decrease in serum T4 level and Cd-induced increase in serum TSH level. Se and Zn can have a synergistic role against Cd-induced thyroid dysfunction.     

Comparative 113Cd-n.m.r. studies on rabbit 113Cd7-, (Zn1,Cd6)- and partially metal-depleted 113Cd6-metallothionein-2a.

Rabbit 113Cd7-metallothionein-2a (MT) contains two metal-thiolate clusters of three (cluster B) and four (cluster A) metal ions. The 113Cd-n.m.r. spectrum of 113Cd6-MT, isolated from 113Cd7-MT upon treatment with EDTA, is similar to that of 113Cd7-MT, but the cluster B resonances are lower in intensity, suggesting its co-operative metal depletion. (Zn1,113Cd6)-MT, formed upon addition of the Zn(II) ions to 113Cd6-MT, shows 113Cd-n.m.r. features characteristic of cluster B populations containing both Cd(II) and Zn(II) ions. The overall intensity gain of the mixed cluster B resonances per Cd as to those in 113Cd6- and 113Cd7-MT suggests a stabilization effect of the bound Zn(II) ions upon the previously established intramolecular 113Cd exchange within this cluster.     

Cadmium and zinc in plants and soil solutions from contaminated soils

In an experiment using ten heavy metal-contaminated soils from six European countries, soil solution was sampled by water displacement before and after the growth of radish. Concentrations of Cd, Zn and other elements in solution (K, Ca, Mg, Mn) generally decreased during plant growth, probably because of uptake by plants and the subsequent redistribution of ions onto soil exchange sites at lower ionic strength. Speciation analysis by a resin exchange method showed that most Cd and Zn in non-rhizosphere solutions was present as Cd2+ and Zn2+, respectively. The proportion of free ions was slightly lower in rhizosphere solutions, mainly due to an increase in dissolved organic carbon during plant growth. Solution pH increased during plant growth, although the bulk soil pH generally remained constant. Cd concentrations in leaves and tubers were more closely correlated with their total or free ionic concentrations in rhizosphere solutions (adjusted R2 0.90) than with their concentrations in soils (adj. R2 0.79). Cd concentrations in non-rhizosphere solutions were only poorly correlated with Cd concentrations in leaves and tubers. In contrast to Cd, there were no soil parameters that individually predicted Zn concentrations in leaves and tubers closely. However, multiple correlation analysis (including Zn concentrations in rhizosphere solutions and in bulk soils) closely predicted Zn concentrations in leaves and tubers (adj. R2 = 0.85 and 0.70, respectively). This suggests that the great variability among soils in the solubility of Zn affected the rate of release of Zn into solution, and thus Zn uptake. There was no such effect for Cd, for which solubility varied much less. Furthermore, the plants may have partly controlled Zn uptake, as they took up relatively less at high solution concentrations of Zn.Free ionic concentrations in soil solution did not predict concentrations of Cd or Zn in plants better than their total concentrations in solution. This suggests that with these soils, analysis of Cd and Zn speciation is of little practical importance when their bioavailability is assessed.     

Effects of land irrigation with partially-treated wastewater on Frankia survival and infectivity

Phytoextraction of Cd by some populations of Thlaspi caerulescens which have the ability to co-hyperaccumulate Cd and Zn requires information about the distribution of both metals within the plant at the organ-level. This work was conducted to determine whether the distribution and solubility of Cd and Zn in Thlaspi caerulescens are affected by the age of plant and organ, and whether Cd and Zn have a common distribution in the plant in soils contaminated by both metals. A series of pot experiments were conducted where a Cd- and Zn-hyperaccumulating population was grown on soils contaminated by Cd and Zn. Temporal changes in metal concentration of roots and of shoots was recorded, along with the water and CaCl₂ solubility of metals in the plant organs. Also, leaves were grouped according to their age and their respective content of Cd and Zn was measured. Both metals were present at higher concentrations in leaves than in roots. The whole-plant content of Zn decreased with time while that of Cd increased or remained unchanged. At harvest, young leaves exhibited higher Cd concentration than older, but the reverse was true for Zn. Both metals were more soluble in dry leaves and senescent leaves than in fresh material, and Zn was more water-soluble than Cd. In conclusion, the distribution of Cd and Zn in the hyperaccumulator T. caerulescensvaried according to the organ and plant age, and Cd and Zn were shown to have a different distribution within the plant.     

Immunological properties of low molecular-weight proteins binding heavy metals in rat kidney and liver

Two homogenous fractions of hepatic metallothioneins ((Cd,Zn) MT-1 and (Cd,Zn) MT-2) and renal metal binding proteins ((Bi,Cu) BP-1 and (Bi,Cu) BP-2) were isolated from rats exposed to heavy metals and specific antisera to them were produced in rabbits.These antisera were tested by immunodiffusion and immunoelectrophoresis for their ability to bind different fractions of hepatic Cd,Zn -metallothionein and renal (Bi,Cu)-, (Hg,Cu)- and (Cd,Cu)-binding proteins. It was found that anti (Bi,Cu) BP antisera did not cross-react with hepatic (Cd,Zn) MT-1 and (Cd,Zn) MT-2. Strong immunological cross-reactions were detected between anti (Bi,Cu) BP antisera and individual forms of (Cd,Cu)-, (Hg,Cu)- and (Bi,Cu)-binding proteins isolated from rat kidneys.     

Comparison of the solution conformations of human [Zn7]-metallothionein-2 and [Cd7]-metallothionein-2 using nuclear magnetic resonance spectroscopy.

The solution structure of native human [Zn7]-metallothionein-2 has been compared with the previously determined structure of human [Cd7]-metallothionein-2. The comparison was based on complete sequence-specific 1H nuclear magnetic resonance assignments for human [Zn7]-metallothionein-2 obtained using the sequential assignment method. The secondary structure was found to be very similar in the [Zn7]- and [Cd7]- forms of the protein. Only seven amide protons in [Zn7]- metallothionein-2 were found to have exchange rates lower than approximately 0.2 min-1 at pH 7.0 and 10 degrees C, which corresponds closely to the results of amide proton exchange studies with the [Cd7]- form of the protein. Finally, the 1H-1H distance constraints determined from nuclear Overhauser enhancement spectroscopy for human [Zn7]-metallothionein-2 were checked for compatibility with the [Cd7]-metallothionein-2 structure. Overall, although no direct method is available for identifying the metal-polypeptide co-ordinative bonds in the Zn(2+)-containing protein, these measurements provided several independent lines of evidence showing that the [Zn7]- and [Cd7]- forms of human metallothionein-2 have the same molecular architecture.