[51] Assays for differentiation of glutathione S-Transferases

Habig, William H.; Jakoby, William B.

doi:10.1016/s0076-6879(81)77053-8

Cited by 2,167 publications

(980 citation statements)

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“…A atividade da enzima GST foi determinada em sangue total, empregando 1-cloro-2,4-dinitrobenzeno e glutationa reduzida como substrato pelo método espectrofotométrico de Habig (1981) 31 , que se baseia na quantificação da formação de 1 µmol do complexo analito GS-DNB/min, monitorada em um comprimento de onda de 340 nm por 60 segundos.…”

Section: Glutationa S-transferase (Gst)unclassified

Avaliação ambiental de BTEX (benzeno, tolueno, etilbenzeno, xilenos) e biomarcadores de genotoxicidade em trabalhadores de postos de combustíveis

Amaral

Carvalho

Pimentel

et al. 2017

Rev. bras. saúde ocup.

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Resumo Introdução: trabalhadores de postos de combustíveis estão expostos às diversas substâncias químicas presentes no ambiente de trabalho, destacando-se entre elas o benzeno, devido às suas propriedades carcinogênicas. Objetivo: avaliar os danos genotóxicos relacionados à exposição ocupacional ao BTEX (benzeno, tolueno, etilbenzeno, xilenos) em trabalhadores de cinco postos de combustíveis do município do Rio de Janeiro, RJ. Metodologia: foram analisadas concentrações de BTEX no ar; atividades das enzimas catalase e glutationa S-transferase; e ensaio cometa em amostras de sangue total de 97 trabalhadores. Resultados: as concentrações de BTEX estavam dentro dos valores preconizados pela NR 15, incluindo Anexo 13-A. Entretanto, uma oscilação nos resultados de ensaio cometa foi observada entre os trabalhadores dos diferentes postos de combustíveis, principalmente em trabalhadores de postos com menores concentrações de benzeno. Discussão: esse resultado está de acordo com a literatura científica atual, que indica uma curva dose-resposta supralinear para o benzeno, observando-se em baixas concentrações um aumento não linear do risco de leucemia, provavelmente relacionado à maior metabolização do benzeno e à maior produção de seus metabólitos tóxicos nessas concentrações. Conclusão: os resultados deste estudo sugerem que a exposição ao BTEX, mesmo em baixas concentrações, contribui para o risco genotóxico à saúde humana.

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Section: Glutationa S-transferase (Gst)unclassified

Avaliação ambiental de BTEX (benzeno, tolueno, etilbenzeno, xilenos) e biomarcadores de genotoxicidade em trabalhadores de postos de combustíveis

Amaral

Carvalho

Pimentel

et al. 2017

Rev. bras. saúde ocup.

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“…One unit of the enzyme activity was defined as the amount of enzyme required to result in a 50% inhibition of the rate of nitro blue tetrazolium reduction measured at 560 nm. GST activity was measured using the method of Habig et al (1974) and Habig and Jakoby (1981) by evaluating the conjugation of GSH (1 mM, Sigma) with the standard model substrate 1-chloro-2, 4-dinitroben-zene (1 mM, Sigma). GPx activity was determined according to Drotar et al (1985), using H 2 O 2 as substrate.…”

Section: Biochemical Analysismentioning

confidence: 99%

Microcystin extracts induce ultrastructural damage and biochemical disturbance in male rabbit testis

Liu¹,

Xie²,

Qiu³

et al. 2009

Environmental Toxicology

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In the present research, the changes of ultrastructures and biochemical index in rabbit testis were examined after i.p. injection with 12.5 lg/kg microcystin (MC) extracts. Ultrastructural observation showed widened intercellular junction, distention of mitochondria, endoplasmic reticulum, and Golgi apparatus. All these changes appeared at 1, 3, and 12 h, but recovered finally. In biochemical analyses, the levels of lipid peroxidation (MDA) and H 2 O 2 increased significantly at 1 h, indicating MC-caused oxidative stress. Finally, H 2 O 2 decreased to the normal levels, while MDA remained at high levels. The antioxidative enzymes (CAT, SOD, GPx, GST) and antioxidants (GSH) also increased rapidly at 1 h, demonstrating a quick response of the defense systems to the oxidative stress. Finally, the activity of CAT, SOD, and GPX recovered to the normal level, while the activity of GST and the concentration of GSH remained at a high level. This suggests that the importance of MCs detoxification by GST via GSH, and the testis of rabbit contained abundant GSH. The final recovery of ultrastructure and some biochemical indexes indicates that the defense systems finally succeeded in protecting the testis against oxidative damage. In conclusion, these results indicate that the MCs are toxic to the male rabbit reproductive system and the mechanism underlying this toxicity might to be the oxidative stress caused by MCs. Although the negative effects of MCs can be overcome by the antioxidant system of testis in this study, the potential reproductive risks of MCs should not be neglected because of their wide occurrence. #

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“…The method for GST assay is a modification of the method described by Habig [29] and has been previously described [27]. Cells were harvested, sonicated, and centrifuged as described previously, and protein concentration determined by the BCA assay.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Cytotoxicity and mutagenicity of dibenzo[a,l]pyrene and (±)-dibenzo[a,l]pyrene-11,12-dihydrodiol in V79MZ cells co-expressing either hCYP1A1 or hCYP1B1 together with human glutathione-S-transferase A1

Kushman

Kabler

Ahmad

et al. 2007

Mutation Research/Fundamental and Molecular Mechanisms of Mutag

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We have used V79MZ hamster lung fibroblasts stably transfected with human cytochrome P450-1A1 (hCYP1A1; cell line designated V79MZh1A1) or P450-1B1 (hCYP1B1; cell line designated V79MZh1B1) alone, or in combination with human GST alpha-1 (hGSTA1), in order to examine GST protection against cytotoxicity and mutagenicity of dibenzo [a,l]pyrene (DBP) and the intermediate dihydrodiol metabolite (+/−)- . At comparable expression levels of hCYP1A1 and hCYP1B1, both DBP and DBPD were more cytotoxic in V79MZ1A1 (IC 50 = 2.7 nM and 0.7 nM, respectively) than in V79MZh1B1 (IC 50 = 6.0 nM and 4.8 nM, respectively). In contrast, both DBP and DBPD were 2-fold to 4-fold more mutagenic in V79MZh1B1 than in V79MZ1A1. Co-expression of hGSTA1 with hCYP1A1 decreased DBP cytotoxicity 2-fold compared to V79MZh1A1 with hCYP1A1 alone, and provided a small, yet still statistically significant, 1.3-fold protection against DBPD. Protection against mutagenicity of these compounds was comparable to that for cytotoxicity in cells expressing hCYP1A1. In V79MZh1B1 cells, co-expression of hGSTA1 conferred up to 5-fold protection against DBP cytotoxicity, and up to 9-fold protection against the (+/−)-DBP-dihydrodiol cytotoxicity relative to the cells expressing hCYP1B1 alone. Co-expression of hGSTA1 also reduced mutagenicity of DBP or its dihydrodiol to a lesser extent (1.3-fold to 1.8-fold) than the protection against cytotoxicity in cells expressing hCYP1B1. These findings demonstrate that the protective efficacy of hGSTA1 against DBP and DBPD toxicity is variable, depending on the compound or metabolite present, the specific cytochrome P450 isozyme expressed, and the specific cellular damage endpoint examined.Keywords glutathione transferase; cytochrome P-450; polycyclic aromatic hydrocarbon; cytotoxicity; mutagenicity; transfection *Corresponding Author: Alan J. Townsend, Ph.D., Biochemistry Department, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem N.C. 27157, Phone (336)-713-7215; FAX (336)-716-7671, E-mail: atown@wfubmc.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. . These reactive products include diol-epoxides with potent mutagenic and carcinogenic activity, some of which are substrates for conjugation by isozymes of the glutathione-S-transferase (GST) superfamily of genes [5]. Conjugation with the nucleophilic thiol group of the tripeptide glutathione (GSH) at the electrophilic centers of activated PAH renders their reactive groups such as epoxides less toxic, more water-soluble and hence more readily excretable [5,6]. Conjugation with GSH also fla...

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[51] Assays for differentiation of glutathione S-Transferases

Cited by 2,167 publications

References 10 publications

Avaliação ambiental de BTEX (benzeno, tolueno, etilbenzeno, xilenos) e biomarcadores de genotoxicidade em trabalhadores de postos de combustíveis

Avaliação ambiental de BTEX (benzeno, tolueno, etilbenzeno, xilenos) e biomarcadores de genotoxicidade em trabalhadores de postos de combustíveis

Microcystin extracts induce ultrastructural damage and biochemical disturbance in male rabbit testis

Cytotoxicity and mutagenicity of dibenzo[a,l]pyrene and (±)-dibenzo[a,l]pyrene-11,12-dihydrodiol in V79MZ cells co-expressing either hCYP1A1 or hCYP1B1 together with human glutathione-S-transferase A1

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