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1.
M W Brown D Shurben J F Solbe A Cryer J Kay 《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1987,87(1):65-69
Two species of coarse fish that are relatively resistant to cadmium poisoning were exposed to sub-lethal concentrations of the metal in their aquarium water. Thus, roach were exposed to cadmium concentrations between 30 and 500 micrograms/l for periods of 14-70 days whereas stone loach were exposed to 1250 micrograms Cd/l for 21-77 days. Under all conditions of exposure, it was found upon analysis of the major organs of accumulation of cadmium in the two species that the toxic metal was sequestered by a single isoform of metallothionein. The amino acid compositions of roach and stone loach metallothionein were determined and found to be similar to those reported for other piscine metallothioneins. The two proteins were found to contain Cd:Zn:Cu in approximate ratios of 4:1:2 per mole of protein. The sequestration of Cd by metallothionein in the two resistant species of fish is contrasted with the situation observed previously in a cadmium-sensitive species, the rainbow trout. 相似文献
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A deficiency of metallothionein, a high-affinity metal-binding protein thought to detoxify cadmium, has been observed in rat and mouse testes, tissues that are highly susceptible to the necrotizing and carcinogenic effects of cadmium. Like the testes, the ovaries undergo a hemorrhagic necrosis when exposed to cadmium, and female Syrian hamsters have recently been shown to be highly susceptible to cadmium. However, the nature of cadmium-binding proteins in the ovary is unknown; thus, this study was undertaken to define the nature of any such proteins in the Syrian hamster ovary. A low molecular weight (Mr) zinc- and cadmium-binding protein was detected in cytosol derived from the ovaries after gel filtration that eluted with a relative elution volume similar to authentic metallothionein. This protein was extractable by heat-treatment and sequential acetone precipitation. When such extracts were further purified with a reverse phase high performance liquid chromatography (HPLC) technique developed for the isolation of metallothionein isoforms, two forms were separated. However, neither of these could be classified as metallothionein on the basis of amino acid composition, since both were particularly low in cysteine, a very common amino acid in metallothionein. The ovarian protein also contained significant amounts of aromatic amino acids, unlike metallothionein--which is devoid of aromatics, and contained much more glutamate than metallothionein. Hamsters were also made resistant to cadmium-induced ovarian necrosis by zinc treatment. Such zinc treatment, however, did not alter levels of this protein, yet caused a marked induction of hepatic metallothionein. Likewise, cadmium treatment did not increase the levels of the ovarian metal-binding protein yet markedly induced hepatic metallothionein.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
4.
Kotrba P Pospisil P de Lorenzo V Ruml T 《Journal of receptor and signal transduction research》1999,19(1-4):703-715
The lamB gene was inserted at with DNA fragments encoding N-terminal beta- and C-terminal alpha-domains of human metallothionein 1A (HMT1A). The hybrid LamB proteins were expressed as full-length products. Virtually whole pool of hybrid LamB proteins was found localized in the outer membrane of E. coli to and cells expressing LamB variants retained sensitivity to lambda phage, indicating their correct folding. Expression of hybrid LamB proteins increased natural ability of E. coli accumulate bivalent heavy metals ions with the highest efficiency observed for cadmium. The order of amount of cadmium accumulated is alpha-domain of HMT1A > HMT1A > beta-domain of HMT1A. This correlates with affinity for cadmium and stability of metallothionein and its individual domains. This confirms suitability of LamB vehicle for surface display of various bioactive molecules and suggests possibility of engineering of cell surface for bioremediation of heavy metals. 相似文献
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Mice were given cadmium chloride orally for 18 weeks, after which they were given two subcutaneous injections of cadmium chloride. Cadmium-and zinc-containing proteins were obtained from livers by ultracentrifugation, Sephadex chromatography and isoelectric focusing. Metallothionein with apI of 4.2 at 8 degrees C was separated from other proteins (possibly including other forms of metallothionein) by the isoelectric procedure. The metallothionein with pI = 4.2 had a high E250, reflecting abundant Cd-SH bonds, in accord with amino acid analysis, which gave a cysteine content of 34.6 residues % and showed an absence of aromatic amino acids. These results indicated a very high purity of the form of metallothionein isolated after isoelectric focusing. 相似文献
6.
The effectiveness of some chelating agents to mobilize cadmium from Chinese hamster ovary cells after chronic exposure (20 hr), as well as from cytosolic metallothionein, was studied. In the first protocol, the most effective substance was 2,3-dimercaptopropanol, followed by 2,3-dimercaptopropane-1-sulfonate and 2,3-dimercaptosuccinic acid, whereas CaNa33-diethylenetriamine pentaacetic acid × 5H2O showed less effect. Simultaneous incubation of cells with cadmium and the chelating agent resulted in a different order of effectiveness: CaNa3 DTPA prevented cadmium uptake almost totally, 2,3-mercaptopropanol by 75% and 2,3-dimercaptopropane-1-sulfonate by 35%. Neither CaNa3-diethylenetriamine pentaacetic acid × 5H2O nor 2,3-dimercaptosuccinic acid had altered the distribution of cadmium between the cytosolic protein fractions after a 2 hr incubation of cells, whereas after this period, 2,3-dimercaptopropanol had removed all cadmium from metallothionein, and 2,3-dimercaptopropane-1-sulfonate about 50%. None of the chelating agents had reduced the amount of Cd bound to high molecular weight proteins. In the cell free system, 2,3-dimercaptopropanol and 2,3-dimercaptopropane-1-sulfonate were equally effective and removed all cadmium from metallothionein within ten minutes. CaNa3-diethylenetriamine pentaacetic acid × 5H2O, however, even after 60 min, had removed only 50% of the cadmium. The remaining cadmium was found distributed to the high molecular weight and lower molecular weight protein fractions.Abbreviations BAL
2,3-dimercaptopropanol
- CHO
Chinese hamster ovary cells
- DMPS
2,3-dimercaptopropane-1-sulfonate
- DMSA
2,3-dimercaptosuccinic acid
- DTPA
CaNa3-diethylenetriaminepentaacetic acid × 5 H2O
- HMW proteins
high molecular weight proteins
- MT
metallothionein 相似文献
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Rainbow trout developed elevated hepatic metallothionein concentrations after 4 weeks in a solution containing zinc, copper and cadmium in a fixed ratio of 400:20:1. Resistance to a combination of these metals increased in proportion to the concentration to which they were exposed for 4 weeks. The concentration of copper but not zinc or cadmium in the low molecular weight proteins separated by gel filtration was related to the concentrations of metallothionein present. The combined toxicity of the metals in the mixture was additive. 相似文献
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The absorption, circular dichroism (CD) and magnetic circular dichroism (MCD) spectra of native rat liver and crab (Scylla serrata) Cd,Zn-metallothionein have been measured and the data are compared. The MCD data indicate that there are close similarities in the geometries of the cadmium-binding sites in both of these proteins; however, the CD spectra are quite different for the rat liver and crab proteins. The CD spectrum for the crab metallothionein is unlike any previously reported for a cadmium-containing metallothionein. This suggests that the CD spectrum is sensitive to the different bridging pattern used in the binding sites in the crab compared with the rat-liver metallothionein. Cadmium binding to the metal-free metallothionein is demonstrated for both proteins and it is shown that there are only minor structural differences between the native and remetallated proteins. The structural changes that occur near to the cadmium-binding sites during cadmium loading to the native proteins have been followed using absorption and CD spectroscopy. Marked changes are observed in the CD spectrum which can be associated with a two-phase reaction: initially Zn2+ is displaced by the Cd2+, then at higher concentrations of Cd2+ the tetrahedral geometry of the Cd2+-binding sites is lost as more Cd2+ is bound using the same thiolate groups. While this latter reaction results in considerable change to the CD spectrum, only minor changes are observed in the absorption spectrum. A significant red shift is observed in the S leads to Cd charge transfer transition region of the MCD spectrum (230-270 nm) following both cadmium loading of native rat liver, Cd,Zn-metallothionein and the metallation of metal-free metallothionein with cadmium. There are two contributions to this effect in Cd,Zn-metallothionein: (i) there is a S leads to Zn band underlying the S leads to Cd band; and (ii) the occupation of zinc sites by cadmium changes the energy of the S leads to Cd transition. 相似文献
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- 1.1. Metal binding proteins with mol. wt in the range 10,000 were isolated from liver and kidney of four different marine species by column chromatography using gel filtration and ionic exchange resins. The proteins were purified with respect to copper, cadmium and zinc. Amino acid analysis was performed on each of the purified proteins.
- 2.2. The amino acid compositions of these metal-binding proteins isolated from fish liver and crab hepatopancreases were very similar to that of metallothionein from rat tissues. Other metal-binding proteins were purified from sea lion kidney and liver, whale liver, crab tissues, and fish liver with amino acid composition similar to rat liver copper chelatin.
- 3.3. Even though some of these metal-binding proteins were found with mol. wt and amino acid compositions similar to rat metallothionein or Cu-chelatin, there were some metal-binding proteins present which appear to be unique to marine animals.
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The induction capacity of dexamethasone, a synthetic glucocorticoid, for the synthesis of metallothionein was about the same as that of 3-aminobenzamide, which is an inhibitor of ADP-ribosylation of chromosomal proteins, in cultured mouse mammary tumor cells. Both inductions of metallothionein were temporally correlated with a decrease in the amount of endogenous poly (ADP-ribose) on nonhistone high-mobility-group 14 and 17 proteins. In contrast, the extent of cadmium-induced metallothionein synthesis was 2-3-times that of dexamethasone or 3-aminobenzamide. However, cadmium had essentially no effect on de-ADP-ribosylation of these proteins. 相似文献
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The absorption, circular dichroism (CD) and magnetic circular dichroism (MCD) spectra of native rat liver and crab (Scylla serrata) Cd,Zn-metallothionein have been measured and the data are compared. The MCD data indicate that there are close similarities in the geometries of the cadmium-binding sites in both of these proteins; however, the CD spectra are quite different for the rat liver and crab proteins. The CD spectrum for the crab metallothionein is unlike any previously reported for a cadmium-containing metallothionein. This suggests that the CD spectrum is sensitive to the different bridging pattern used in the binding sites in the crab compared with the rat-liver metallothionein. Cadmium binding to the metal-free metallothionein is demonstrated for both proteins and it is shown that there are only minor structural differences between the native and remetallated proteins. The structural changes that occur near to the cadmium-binding sites during cadmium loading to the native proteins have been followed using absorption and CD spectroscopy. Marked changes are observed in the CD spectrum which can be associated with a two-phase reaction: initially Zn2+ is displaced by the Cd2+, then at higher concentrations of Cd2+ the tetrahedral geometry of the Cd2+-binding sites is lost as more Cd2+ is bound using the same thiolate groups. While this latter reaction results in considerable change to the CD spectrum, only minor changes are observed in the absorption spectrum. A significant red shift is observed in the S → Cd charge transfer transition region of the MCD spectrum (230–270 nm) following both cadmium loading of native rat liver, Cd,Zn-metallothionein and the metallation of metal-free metallothionein with cadmium. There are two contributions to this effect in Cd,Zn-metallothionien: (i) there is a S → Zn band underlying the S → Cd band; and (ii) the occupation of zinc sites by cadmium changes the energy of the S → Cd transition. 相似文献
13.
Isolation of a novel metal-binding protein from rat testes. Characterization and distinction from metallothionein 总被引:2,自引:0,他引:2
This study was undertaken to further establish the nature of the low molecular weight metal-binding proteins in rat testes. In all cases, control testes were compared to livers of zinc-treated rats, which are known to contain high concentrations of metallothionein. Gel filtration of testicular and hepatic cytosol revealed a major zinc- and/or cadmium-binding protein in the low molecular weight range in both tissues. This protein could be partially purified from either source by a combination of heat treatment and sequential acetone precipitation. When such partially purified preparations were further fractionated by high performance liquid chromatography using a linear gradient of 25%-40% acetonitrile in 0.1% trifluoroacetic acid, two major forms with similar retention times were seen in each tissue. The utility of this high performance liquid chromatography system for separating isoforms of metallothionein was verified by separation of commercially available purified rabbit hepatic metallothionein into a total of five separate forms. Amino acid analysis of the two proteins derived from rat liver was consistent with the known amino acid composition of metallothionein. However, the two testicular forms separated by high performance liquid chromatography were notably different in amino acid composition from metallothionein, with a distinctly lower content of cysteine. These results indicate that the major low molecular weight cadmium/zinc-binding proteins in rat testes are not metallothioneins. 相似文献
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《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》2000,125(1):17-24
Metallothionein is considered to be a potential biomarker for heavy metal exposure in the terrestrial environment. However, limited information is available on metallothioneins from insects, a major class of terrestrial invertebrates. In this study we have quantified metallothioneins in the springtail Orchesella cincta by determining metallothionein-bound cadmium after separation of these proteins using gel filtration and reversed phase chromatography from total body homogenates of animals dietary exposed to different concentrations of cadmium. Furthermore, we have studied in more detail where cadmium and metallothionein-bound cadmium is located within this animal. The concentration of metallothionein-bound cadmium increases with the exposure concentration in the same way as the total internal concentration. Both reach a plateau at an exposure concentration of approximately 1.0 μmol Cd/dry food. Cadmium is primarily located within the gut of O. cincta and isolation of metallothionein from this organ gives results identical to isolations from total bodies. Based on this results an estimation of the metallothionein level at the highest exposure concentration results in a concentration of about 115 μg metallothionein/g fresh gut. The O. cincta metallothionein gives the possibility of using this protein as a biomarker for heavy metal exposure in soil insects. 相似文献
15.
Properties of cadmium-binding proteins in rat testes. Characteristics unlike metallothionein.
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Since the exposure of rats to cadmium causes zinc to accumulate in metallothionein in liver and kidney but not in a similar protein in the testes, the properties of the low-Mr cadmium-binding proteins were investigated in rat testes. Weanling rats that had been given dietary cadmium for 6 weeks were injected with 109CdCl2 and subsequently killed, and the 109Cd-labelled low-Mr proteins from testes were purified. The pooled low-Mr cadmium-containing fractions from the gel-filtration (Sephadex G-75) columns were eluted through DEAE-Sephacel columns, yielding two peaks. Each of the individual peaks from this Sephacel column was further purified by rechromatography on DEAE-Sephacel and on Bio-Gel P-10 columns. Amino acid analysis of the two purified proteins revealed a low cysteine (about 3%) content, with aspartate, glutamate and glycine as the predominant amino acids. Thus these low-Mr cadmium-binding proteins induced by cadmium in rat testes do not appear to be metallothionein. 相似文献
16.
Smoryzanova OA Synzynys BI Danilin IA Baranova OA 《Radiatsionnaia biologiia, radioecologiia / Rossi?skaia akademiia nauk》2004,44(2):179-182
It was confirmed that the survival of gamma-exposed mice became higher after preliminarily subcutaneous injections of cadmium chloride. It is supposed that radioresistance of mice is connected with the induction of metallothionein proteins by cadmium chloride. 相似文献
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Cadmium is a toxic metal that induces the expression of metallothionein genes in many tissues and that binds avidly to metallothionein, a soluble transition metal binding protein. The present study examined the temporal pattern and magnitude of accumulation of metallothionein mRNA in liver of C57BL/6J mice of various ages treated with cadmium. In adult female mice, accumulation was dependent on the dosage level of cadmium and related to the concentration of this metal in liver. The accumulation of metallothionein mRNA in liver depended on age at exposure to cadmium. Intraperitoneal administration of 2 mg of cadmium per kg provoked small increases (two- to threefold) in levels of metallothionein mRNA in livers of 7- and 14-day-old mice. In contrast, cadmium treatment of 28- and 56-day-old mice resulted in 12- to 19-fold increases in levels of metallothionein mRNA in liver with maximum increases occurring 3 to 4 hr after treatment. Because similar patterns for the accumulation of cadmium of liver were found in 7-, 28-, and 56-day-old mice, observed age-dependent differences in induction of metallothionein mRNA in liver were probably not due to differences in the accumulation of cadmium in this organ. Taken together, these data suggest that tissue-specific factors controlling the expression of metallothionein genes may account for developmental variation in the inducibility of these genes by cadmium. Ontogenic variation in accumulation of metallothionein mRNA after cadmium treatment may be a factor in developmental variation in the acute lethality of cadmium in C57BL/6J mice. 相似文献
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The main binding protein for 109Cd was metallothionein after in vitro incubation of various tissue cytosol preparations obtained from rats supplemented with zinc. The exception was heart cytosol where the label was associated with higher molecular weight proteins. The metallothionein-bound 109Cd was sensitive to trithiomolybdate and moved too higher molecular weight proteins, presumably because of the creation of new stronger ligands by the association of thiomolybdate with these proteins. The 109Cd binding was affected by selenate, selenite, and selenide while molybdate, sulphate, and thiosulphate were ineffective. It is proposed that thiomolybdates should be investigated for use in the therapy of in vivo cadmium toxicity because they can remove the accumulated metal from metallothionein. 相似文献
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J Overnell 《Comp. Biochem. Physiol. C, Comp. Pharmacol. Toxicol.》1984,77(2):237-243
In the presence of 2-mercaptoethanol DEAE cellulose chromatography of desalted hepatopancreas cytosols of crabs rich in both copper and cadmium yielded three peaks, probably all due to metallothioneins, with very different copper to cadmium ratios. Addition of a copper glycine complex to a desalted crab cytosol containing cadmium metallothionein gave a chromatogram similar to that obtained by in vivo administration of copper and cadmium. The chromatographic distribution of the metals between the different charge-forms in the distal and proximal portions of the tubules was basically similar although the concentrations of the metals were different. 相似文献