Isolation and Some Properties of Human Metallothionein<sup>*</sup>

Pulido, Pablo; Kägi, Jeremias H. R.; Vallée, Bert L.

doi:10.1021/bi00869a046

Cited by 324 publications

(80 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the human, metallothionein was isolated from kidney (16) and from liver (1). The metallic constituents of the adult hepatic metallothionein is almost exclusively zinc, while the adult renal protein contains nearly equal amounts of cadmium and zinc, as well as trace amounts of copper and/or mercury in some cases (16).…”

Section: Discussionmentioning

confidence: 99%

“…The metallic constituents of the adult hepatic metallothionein is almost exclusively zinc, while the adult renal protein contains nearly equal amounts of cadmium and zinc, as well as trace amounts of copper and/or mercury in some cases (16). On the other hand, the search for copper-containing proteins in newborn liver, an organ extremely rich in copper, was initiated by Porter et al (14).…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, the search for copper-containing proteins in newborn liver, an organ extremely rich in copper, was initiated by Porter et al (14). Mitochondrocuprein, a particulate copper-protein, was fractionated in 1964 (16). Recently a copperbinding low molecular weight protein was purified after homogenization of human fetal liver (17).…”

Section: Discussionmentioning

confidence: 99%

“…Human metallothioneins were isolated from adult liver (1) and kidney (16). The hepatic protein contains almost exclusively zinc, but also both cadmium and zinc in about equal proportions are bound to renal metallothionein.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Immunohistological Studies of Metallothionein

Kojima

Hamashima

1980

Acta Histochem. Cytochem

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Immunohistological Studies of Metallothionein

Kojima

Hamashima

1980

Acta Histochem. Cytochem

View full text Add to dashboard Cite

“…Another feature of metallothionein is its characteristic low absorption at 280 nm, due to a lack of aromatic amino acids (10), and its relatively high absorption at 250 nm, attributed to metalmercaptide chromophores (18,25). When the material eluting at high ionic strength from the DEAE-cellulose column was desalted, concentrated, and brought to 50 mm Tris (pH 7.8), it yielded a UV-absorption spectrum characteristic of metallothionein (Fig.…”

mentioning

confidence: 99%

Partial Characterization of a Cadmium-binding Protein from the Roots of Cadmium-treated Tomato

1980

View full text Add to dashboard Cite

A Cd-binding protein has been isolated from the roots of Cd-treated tomato plants cv. Rutgers. Almost all the Cd from a high-speed supernatant fraction was recovered in a 10,000-dalton fraction from a gel filtration column coincident with 250-nanometer absorbing material. DEAE-cellulose chromatography of this 10,000-dalton material yielded one major component, which eluted at 0.34 molar NaCI, had an absorption spectrum characteristic of metaDlothioein, and showed absorption changes upon acidification typical of metallothionein. Although the Cd-binding protein did not behave like metallothioneins from animal sources during gel electrophoresis at pH 8.9, a single band containing Cd and staining with Coomassie brilliant blue could be detected following electrophoresis at pH 6.9. Synthesis of the Cd-binding protein appeared to be "induced" by treatment of the plants with Cd2". Recent concern over Cd as an environmental contaminant (23) has stimulated research on the response of plants to Cd (7,15,16). Although dose-response data have been obtained for many plant species, little is known concerning the biochemical mechanisms of Cd toxicity or tolerance (19,22,23 Preparation of Cd-Thionein. The high-speed supernatant was fractionated via gel filtration using Sephadex G-50 fine (Pharmacia Fine Chemicals). The column was equilibrated and the sample eluted with 50 mM Tris (pH 7.8). The appropriate fractions were combined and then subjected to DEAE-cellulose chromatography (Whatman DE52). The DEAE-cellulose column was equilibrated with 50 mM Tris (pH 7.8) and, after sample application, washed with the same buffer. The sample was eluted with 200 ml of a linear gradient of 0.20 to 0.40 M NaCl in 50 mm Tris (pH 7.8). The appropriate fractions were combined and then desalted with coarse Sephadex G-25 using deionized H20 as eluant. The sample then was concentrated by ultrafiltration using an Amicon UM-2 filter. Column dimensions are indicated in the figure legends.Polyacrylamide Gel Electrophoresis. Polyacrylamide gel electrophoresis was via tube gels, 10 cm long and 0.5 cm in diameter. The running gel was 12% acrylamide (0.4% bisacrylamide) and contained 0.03% (v/v) TEMED5, 0.0 175% (w/v) ammonium persulfate, and 0.38 M bis-Tris adjusted to pH 6.9 with HC1. The stacking gel was 2.5% acrylamide (0.625% bisacrylamide) and contained 23% (w/v) sucrose, 0.0625% (v/v) TEMED, 0.07% (w/v) ammonium persulfate, and 62.5 mM bis-Tris adjusted to pH 5.9 with HC1. The reservoir buffer was 20 mm bis-Tris adjusted to pH 6.5 with glutamic acid. Electrophoresis was at room temperature with DNP-aspartate as "tracking dye." Prior to electrophoresis, samples were mixed with an equal volume of sample buffer consisting of 32% (w/v) sucrose and 100 mm bis-Tris adjusted to pH 5.9 with HC1. Duplicate gels were run. One was stained at 5 Abbreviations: bis-Tris: bis(2-Hydroxyethyl)imino-tris(Hydroxymethyl) methane; DNP-aspartate: 2,4-dinitrophenyl aspartate;

show abstract

Quantitative determination of Cu-thionein from human fluids with application of solid-phase extraction on covalent affinity chromatography with thiol-disulphide interchange support

Kabziński

2000

Biomed. Chromatogr.

View full text Add to dashboard Cite

The aim of our investigation was to carry out quantitative isolation of Cu-thionein (Cu-Th) in human body fluids (urine, plasma and breast milk) in order to determine the level of exposure to heavy metals. In the experiment covalent affinity chromatography with thiol-disulphide interchange gel (CAC-TDI) was used as a solid phase extraction (SPE) support for preconcentration of Cu-thionein (Cu-Th) protein and Cu bonded with MT from water and human fluids samples. The present paper is a continuation of early experiments on the quantitation of Hg-thionein (Hg-Th), Cd-thionein (Cd-Th) and Zn-thionein (Zn-Th) in human body fluids such as urine, plasma and breast milk.

show abstract

Isolation and Some Properties of Human Metallothionein^*

Cited by 324 publications

References 32 publications

Immunohistological Studies of Metallothionein

Immunohistological Studies of Metallothionein

Partial Characterization of a Cadmium-binding Protein from the Roots of Cadmium-treated Tomato

Quantitative determination of Cu-thionein from human fluids with application of solid-phase extraction on covalent affinity chromatography with thiol-disulphide interchange support

Contact Info

Product

Resources

About

Isolation and Some Properties of Human Metallothionein*

Cited by 324 publications

References 32 publications

Immunohistological Studies of Metallothionein

Immunohistological Studies of Metallothionein

Partial Characterization of a Cadmium-binding Protein from the Roots of Cadmium-treated Tomato

Quantitative determination of Cu-thionein from human fluids with application of solid-phase extraction on covalent affinity chromatography with thiol-disulphide interchange support

Contact Info

Product

Resources

About

Isolation and Some Properties of Human Metallothionein^*