Isolation and Characterization of Cadmium-Binding Protein From Rat Testes

Singh, Kartar; Nath, R.; Chakravarti, R. N.

doi:10.1530/jrf.0.0360257

Cited by 21 publications

(9 citation statements)

References 4 publications

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“…The TMBP does display several striking similarities to MT. In agreement with several previous studies, the present results demonstrate that the TMBP behaves similarly to MT on gel-filtration chromatography (Nordberg, 1971;Singh et al, 1974;Chen & Ganther, 1975;Brady & Webb, The gels were scanned at 650nm and relative mobility (Bromphenol Blue).…”

Section: Discussionsupporting

confidence: 91%

“…Among several other differences, the TMBPs also contained approximately half the amount of serine and lysine found in the two forms of MT. The presence of aromatic amino acids in TMBP may account for the u.v.-spectral differences between it and MT found in present and previous studies (Chen et al, 1972;Singh et al, 1974).…”

Section: Discussionsupporting

confidence: 48%

“…Chen et al (1972) and Singh et al (1974) have reported that the u.v.-absorption spectra of the testicular MT-like protein differs markedly from that of hepatic or renal MT. It has also been reported that, unlike hepatic MT, the testicular MT-like protein does not accumulate Zn after Cd exposure (Whanger et al, 1980).…”

Section: Vol 220mentioning

confidence: 99%

“…Work by Nordberg (1971), Chen et al (1974, Singh et al (1974) and Chen & Ganther (1975) have provided evidence for rodent testicular MT after Cd treatment. Webb (1975) has also detected a Znbinding MT-like protein in untreated rat testes.…”

mentioning

confidence: 98%

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Cadmium-binding proteins of rat testes. Characterization of a low-molecular-mass protein that lacks identity with metallothionein

Waalkes¹,

Chernoff²,

Klaassen³

1984

Biochemical Journal

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Cadmium-binding proteins in the cytosol of testes from untreated rats were separated by Sephadex G-75 gel filtration. Three major testicular metal-binding proteins (TMBP), or groups of proteins, with relative elution volumes of approx. 1.0 (TMBP-1), 1.7 (TMBP-2) and 2.4 (TMBP-3) were separated. Elution of Zn-binding proteins exhibited a similar pattern. TMBP-3 has previously been thought to be metallothionein (MT), and hence this protein was further characterized and compared with hepatic MT isolated from Cd-treated rats. Estimation of M, by gel filtration indicated a slight difference between MT (Mr 10000) and TMBP-3 (Mr 8000). Two major forms of MT (MT-I and MT-II) and TMBP-3 (TMBP-3 form I and TMBP-3 form II) were obtained after DEAE-Sephadex A-25 anion-exchange chromatography, with the corresponding subfractions being eluted at similar conductances. Non-denaturing polyacrylamide-gel electrophoresis on 7% acrylamide gels indicated that the subfractions of TMBP-3 had similar mobilities to those of the corresponding subfractions of MT. However, SDS (sodium dodecyl sulphate)/12% (w/v)-polyacrylamide-gel electrophoresis resulted in marked differences in migration of the two corresponding forms of MT and TMBP-3. Co-electrophoresis of MT-II and TMBP-3 form II by SDS/polyacrylamide-gel electrophoresis revealed two distinct proteins. Amino acid analysis indicated much lower content of cysteine in the testicular than in the hepatic proteins. TMBP-3 also contained significant amounts of tyrosine, phenylalanine and histidine, whereas MT did not. U.v.-spectral analysis of TMBP-3 showed a much lower A250/A280 ratio than for MT. Thus this major metal-binding protein in testes, which has been assumed to be MT is, in fact, a quite different protein.Metallothioneins (MT) are metal-binding proteins containing high levels of cysteine (approx. 18

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 48%

Section: Vol 220mentioning

confidence: 99%

mentioning

confidence: 98%

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Cadmium-binding proteins of rat testes. Characterization of a low-molecular-mass protein that lacks identity with metallothionein

Waalkes¹,

Chernoff²,

Klaassen³

1984

Biochemical Journal

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“…Transgenic mice overexpressed MT-1 in the testes was still not sufficient to protect the tissue from Cd-induced testicular damage (Dalton et al, 1996). One plausible explanation could be that Cd cannot be easily distributed to the testes and is present at levels much lower than in other tissues such as livers (Singh et al, 1974). However, an acute dose of Cd has been shown to produce testicular damage within one day after intraperitoneal administration.…”

Section: Discussionmentioning

confidence: 99%

Tissue-Dependent Induction of Heme Oxygenase-1 and Metallothionein-1/2 by Methyl Methanesulfonate

Ashino¹,

Ozawa

Numazawa³

et al. 2003

J. Toxicol. Sci.

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-Methyl methanesulfonate (MMS), a methylating agent, is known to be a genotoxicant in testis. The purpose of this study was to investigate roles of oxidative stress-responsive proteins, heme oxygenase-1 (HO-1) and metallothionein-1/2 (MT-1/2), in genotoxicity of MMS. Cadmium, a potent genotoxicity inducer, induced HO-1 and MT-1/2 in rat livers and kidneys. Then we comparatively investigated MMS-induced HO-1 and MT-1/2 in rat livers, kidneys and testes. We found that a single administration of MMS (40 mg/kg) resulted in the induction of MT-1/2 mRNA in the liver, but not HO-1 mRNA, reaching maximum level at 6 hr and returning to the control levels by 24 hr. Interestingly, MMS induced both HO-1 and MT-1/2 mRNAs in the kidney. In contrast, MMS induced HO-1 mRNA, but not MT-1/2 mRNA in the testis. Since HO-1 and MT-1/2 have been recognized to respond to various oxidative stimuli, we further examined the inducing effect of MMS on these two proteins. MMS at dosages of 20 to 40 mg/kg for 2 consecutive weeks induced HO-1 mRNA (123 to 187% of the control) and protein (274 to 404% of the control) in rat testes. However, MT-1/2 mRNA was not induced by MMS administration, although a high level of expression was observed in comparison with the liver and kidney. These findings suggest that MMS induces HO-1 and/or MT-1/2 mRNA and its protein tissue-dependently, and the heme catabolites by HO-1 in the testis may contribute in some manner to its genotoxicity.

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Functional integrity of metallothionein genes in testicular cell lines

Wang¹,

Chen²,

Lin³

1994

J of Cellular Biochemistry

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The presence and inducibility of the major cadmium (Cd) chelating protein metallothionein (MT) in testicular cells has been controversial. In this study, the induction and production of MT in testicular cells were studied using mouse Leydig and Sertoli cell lines. Metal accumulation was studied by subjecting the cells to increasing levels of Cd. The presence of transcription factors for MT synthesis was analyzed by transfecting the cells with a reporter gene under the control of the MT promoter. The dose- and time-dependent induction of MT were conducted by Northern analyses. Expression of MT genes occurred in both Leydig and Sertoli cells. To avoid cross hybridization of the MT probe with mRNAs encoding testicular metal binding proteins and to investigate the integrity of MT mRNA, isoMT mRNA identification and primer extension experiments were performed. Those studies show that the induced mRNA indeed encodes MT. The biosynthesis of MT was confirmed by following 35S-cysteine incorporation into the protein. Finally, cadmium tolerance of testicular cells is compared with that of fibroblast cells. By these studies, we conclude that the MT genes are functional and inducible in testicular cells.

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Isolation and Characterization of Cadmium-Binding Protein From Rat Testes

Cited by 21 publications

References 4 publications

Cadmium-binding proteins of rat testes. Characterization of a low-molecular-mass protein that lacks identity with metallothionein

Cadmium-binding proteins of rat testes. Characterization of a low-molecular-mass protein that lacks identity with metallothionein

Tissue-Dependent Induction of Heme Oxygenase-1 and Metallothionein-1/2 by Methyl Methanesulfonate

Functional integrity of metallothionein genes in testicular cell lines

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