Purification and characterization of three distinct glutathione transferases from mouse liver

Warholm, Margareta; Jensson, Helgi; Tahir, Muhammad Aslam; Mannervik, Bengt

doi:10.1021/bi00362a020

Cited by 81 publications

(73 citation statements)

References 30 publications

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“…Notwithstanding the difficulties in identifying classes of glutathione transferase in different species, it was recently noted (4) that the amino-terminal amino acid sequences of two homologous mouse transferases (15) showed extensive similarities to those of two homologous rat transferases (16). This finding prompted studies of the glutathione transferase isoenzymes in mouse liver (17) and in human tissues (7), as well as an attempt at interspecies classification ofthe multiple forms of the enzyme (18). The present report correlates enzymatic and structural distinctions and shows that glutathione transferases from several species can be assigned to three classes comprising isoenzymes with common characteristic properties.…”

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confidence: 57%

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Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties.

Mannervik¹,

Ålin²,

Guthenberg³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

947

518

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The major isoenzymes of cytosolic glutathione transferase (EC 2.5.1.18) from rat, mouse, and man are shown to share structural and catalytic properties that can be used for species-independent classification. Rat, mouse, and human isoenzymes were grouped with respect to aminoterminal amino acid sequences, after correlation of seven structures analyzed in the present investigation with structures determined earlier. The isoenzymes were also characterized by substrate specificities and sensitivities to inhibitors, and the data were subjected to pattern recognition analysis. In addition, the various isoenzymes were tested for cross-reactivity by immunoprecipitation with antibodies raised against rat and human transferases. The different types of data were clearly correlated and afforded an unambiguous division of the isoenzymes into three classes named alpha, mu, and pi. Each of the three mammalian species studied contains at least one isoenzyme of each class. It is suggested that the similarities of the isoenzymes in a class reflect evolutionary relationships and that the classification applies generally.

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confidence: 57%

“…Glutathione transferases from human placenta (19), human liver (20), rat liver (21), rat kidney (14), rat testis (13), and mouse liver (17) were purified to homogeneity by procedures as indicated. Antibodies to purified transferases were raised in rabbits.…”

mentioning

confidence: 99%

Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties.

Mannervik¹,

Ålin²,

Guthenberg³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

947

518

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“…On the other hand, GSTs purified from rat and mouse livers exhibited selenium-independent glutathione peroxidase activity (19,22), indicating that the properties of catalytic sites between mammalian and microbial enzymes may be different. The cDNAs responsible for GSTs of corn (GST III [14]) and rat (Ya and Yc [21]) have been cloned, sequenced, and expressed in E. coli cells.…”

Section: Resultsmentioning

confidence: 99%

Purification and some properties of glutathione S-transferase from Escherichia coli B

et al. 1989

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Glutathione S-transferase was purified approximately 2,300-fold from cell extracts of Escherichia coli B with a 7.5% activity yield. The molecular weight of the enzyme was 45,000, and the enzyme appeared to consist of two homogeneous subunits. The enzyme was almost specific to 1-chloro-2,4-dinitrobenzene (K,. 1.43 mM) and glutathione (K,,, 0.33 mM). The optimal pH and optimal temperature for activity were 7.0 and 50°C, respectively, and the enzyme was stable from pH 5 to 11. The activity of the enzyme for 1-chloro-2,4-dinitrobenzene (3.2 ,umol/min per mg of protein) was significantly lower than those of the enzymes from mammals, plants, and fungi.Glutathione S-transferases (GSTs) constitute an important class of detoxifying enzymes. The enzyme catalyzes the conjugation of glutathione (GSH) with various compounds having electrophilic and/or hydrophobic sites (6). The enzyme is thought to protect the cells against foreign compounds such as pesticides, drugs, and carcinogens (7). The enzymes have been extensively purified from mammals such as human (8), mouse (12), cattle (3), and rat (21), and their properties have been characterized in detail. However, the data on microbial GSTs are largely lacking, and the enzyme has been purified only from Fusarium oxysporum f. sp. melonis (5), Mucorjavanicus (1), and Tetrahymena thermophila (18). Although attempts to detect GST activity in cell extracts of bacteria (2, 11, 21), yeasts (4, 9), mold (1, 5), and protozoa (18) have been made, these results are fragmentary and not enough to compare the properties of microbial enzymes with those of enzymes from mammals. Here we report the purification and partial characterization of GST from Escherichia coli B.MATERIALS AND METHODS Chemicals. GSH was purchased from Kohjin Co., Ltd., Tokyo, Japan. 1-Chloro-2,4-dinitrobenzene was from Tokyo Kasei Kogyo Co., Ltd., Tokyo, Japan. Glutathione agarose was prepared from AH-Sepharose CL-4B according to the vendor's specifications (Pharmacia).Assays. The activity of GST was assayed essentially by the method of Habig et al. (6). The reaction mixture (1.0 ml) consisted of 0.1 M potassium phosphate buffer (KPB) (pH 7.0), 1.0 mM EDTA, 1.0 mM GSH, 1.0 mM 1-chloro-2,4-dinitrobenzene (CDNB), and enzyme. The enzyme activity was calculated by using a molar extinction coefficient of S-(2,4-dinitrophenyl)glutathione as 9.6 M-1 cm-' at 340 nm and 25°C. One unit of enzyme activity was defined as the amount producing 1 nmol of conjugate of GSH with CDNB per min. Protein was determined by the method of Lowry et al. (13). GSH in cells was determined by the method of Murata et al. (17).Growth experiments. In order to investigate the effect of electrophiles on the formation of GST activity, cells of E. coli B were grown in a test tube containing 5 ml of nutrient medium (1.0% glucose, 0.1% yeast extract, 1.0% peptone, 0.5% meat extract, 0.1% MgSO4-7H20, and 0.5% K2HPO4 * Corresponding author.[pH 7.0]) with reciprocal shaking at 37°C for 20 h. The cells were transferred into a 2-liter Sakaguchi flask containing...

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“…Multiple GST forms have been described in the rat (Habig et al, 1974;Hayes, 1984Hayes, , 1986Kitahara et al, 1984;Tu & Reddy, 1985;Guthenberg et al, 1985), the mouse (Lee et al, 1981;Pearson et al, 1983;Warholm et al, 1986;Hayes et al, 1987) and man (Awasthi et al, 1980;Stockman et al, 1985). Both cytosolic and microsomal GST exist.…”

Section: Introductionmentioning

confidence: 99%

“…Cytosolic and microsomal GST from the mouse have been studied in several laboratories; the physical properties of the enzymes isolated by separate groups of .workers differ (Lee et al, 1981;Lee & McKinney, 1982;Pearson et al, 1983;Morgenstern et al, 1984;Warholm et al, 1986: Hayes et al, 1987. The reasons for the lack of consistency of results from different research groups are not clear, but they suggest that strain-specific variations in mouse GST might occur.…”

Section: Introductionmentioning

confidence: 99%

Sex-specific constitutive expression of the pre-neoplastic marker glutathione S-transferase, YfYf, in mouse liver

McLellan

Hayes

1987

Biochemical Journal

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Hepatic glutathione S-transferase isoenzyme content has been investigated in both sexes of three inbred strains of mice (DBA/2, C3H/He, C57BL6). A polypeptide (Mr 24,800), which is immunologically related to Yf purified from rat lung, was found to be expressed as a major form in all male mouse livers but represented only a minor enzyme form in female mouse liver. Glutathione S-transferases comprising subunits with molecular masses of 25,800 (Ya) or 26,400 (Yb) were present in males and females of the three strains under investigation. Cytosolic isoenzymes from all strains and sexes were purified to apparent homogeneity and no significant inter-strain differences in the properties of the individual forms were observed. In addition, no differences were detected in the microsomal glutathione S-transferase content of the different strains or sexes.

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Purification and characterization of three distinct glutathione transferases from mouse liver

Cited by 81 publications

References 30 publications

Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties.

Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties.

Purification and some properties of glutathione S-transferase from Escherichia coli B

Sex-specific constitutive expression of the pre-neoplastic marker glutathione S-transferase, YfYf, in mouse liver

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