Glutathione S-transferases of human lung: Characterization and evaluation of the protective role of the α-class isozymes against lipid peroxidation

Singhal, Sharad S.; Saxena, Manju; Ahmad, Hassan; Awasthi, Sanjay; Haque, Abida K.; Awasthi, Yogesh C.

doi:10.1016/0003-9861(92)90269-3

Cited by 109 publications

(53 citation statements)

References 40 publications

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“…Although no information is available about the function of this gene, its sequence homology suggests that it may have similar properties to the hydroxyacid-oxidase enzyme in its oxidation of substrates such as long-chain fatty acids. Consistent with this is the finding that GST, an enzyme induced by lipid peroxides (Singhal et al 1992;Khan et al 1995;Tjalkens et al 1998), was increased in the SR-BI transgenics. Furthermore, the oxoglutarate carrier protein, which facilitates the transport of glutathione into the mitochondria (Chen & Lash 1998), was also observed to be decreased in the SR-BI transgenic (adjusted 0.05 <P <0.2).…”

Section: Discussionsupporting

confidence: 76%

Microarray Expression Profiling Identifies Genes with Altered Expression in HDL-Deficient Mice

Callow

Dudoit²,

Gong³

et al. 2000

Genome Research

211

174

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Based on the assumption that severe alterations in the expression of genes known to be involved in high-density lipoprotein (HDL) metabolism may affect the expression of other genes, we screened an array of >5000 mouse expressed sequence tags for altered gene expression in the livers of two lines of mice with dramatic decreases in HDL plasma concentrations. Labeled cDNA from livers of apolipoprotein AI (apoAI)-knockout mice, scavenger receptor BI (SR-BI) transgenic mice, and control mice were cohybridized to microarrays. Two-sample t statistics were used to identify genes with altered expression levels in the knockout or transgenic mice compared with control mice. In the SR-BI group we found nine array elements representing at least five genes that were significantly altered on the basis of an adjusted P value < 0.05. In the apoAI-knockout group, eight array elements representing four genes were altered compared with the control group (adjusted P < 0.05). Several of the genes identified in the SR-BI transgenic suggest altered sterol metabolism and oxidative processes. These studies illustrate the use of multiple-testing methods for the identification of genes with altered expression in replicated microarray experiments.

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Section: Discussionsupporting

confidence: 76%

Microarray Expression Profiling Identifies Genes with Altered Expression in HDL-Deficient Mice

Callow

Dudoit²,

Gong³

et al. 2000

Genome Research

211

174

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“…4-HNE was purchased from Cayman Chemical Co. (Ann Arbor, MI). Dilinoleoylphosphatidylcholine hydroperoxide (PC-OOH), Dilinoleoylphosphatidylethanolamine hydroperoxide (PE-OOH), 9-hydroperoxylinoleic acid (9-LOOH), and 13-hydroperoxylinoleic acid (13-LOOH) were synthesized as described previously (40). Lipid hydroperoxides and 4-HNE were stored at Ϫ70°C under nitrogen atmosphere.…”

Section: Methodsmentioning

confidence: 99%

“…All reagents for SDS-PAGE and Western transfer were purchased from Bio-Rad. The polyclonal antibodies raised against the Alpha, Mu, and Pi class GSTs were the same as those used in our previous studies (40). Human tissue samples were obtained from the autopsy service at the University of Texas Medical Branch with no diagnosed disorders related to investigated organs, and blood was obtained from the University of Texas Medical Branch blood bank.…”

Section: Methodsmentioning

confidence: 99%

“…The BL21 cells were lysed by sonication in 10 mM potassium phosphate buffer, pH 7.0, containing 1.4 mM ␤-mercaptoethanol (buffer A) and centrifuged for 45 min at 28,000 ϫ g at 4°C. The supernatants were collected and subjected to affinity chromatography using GSH linked to epoxy-activated Sepharose 6B (40). The recombinant GSTs were eluted from the GSH affinity resin with 50 mM TrisHCl, pH 9.6, containing 10 mM GSH and 1.4 mM ␤-mercaptoethanol.…”

Section: Methodsmentioning

confidence: 99%

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Role of Glutathione S-Transferases in Protection against Lipid Peroxidation

Yang

Cheng

Singhal

et al. 2001

Journal of Biological Chemistry

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284

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The physiological significance of the selenium-independent glutathione peroxidase (GPx) activity of glutathione S-transferases (GSTs), associated with the major Alpha class isoenzymes hGSTA1-1 and hGSTA2-2, is not known. In the present studies we demonstrate that these isoenzymes show high GPx activity toward phospholipid hydroperoxides (PL-OOH) and they can catalyze GSH-dependent reduction of PL-OOH in situ in biological membranes. A major portion of GPx activity of human liver and testis toward phosphatidylcholine hydroperoxide (PC-OOH) is contributed by the Alpha class GSTs. Overexpression of hGSTA2-2 in K562 cells attenuates lipid peroxidation under normal conditions as well as during the oxidative stress and confers about 1.5-fold resistance to these cells from H 2 O 2 cytotoxicity. Treatment with 30 M H 2 O 2 for 48 h or 40 M PC-OOH for 8 h causes apoptosis in control cells, whereas hGSTA2-2-overexpressing cells are protected from apoptosis under these conditions. In control cells, H 2 O 2 treatment causes an early (within 2 h), robust, and persistent (at least 24 h) activation of JNK, whereas in hGSTA2-2-overexpressing cells, only a slight activation of JNK activity is observed at 6 h which declines to basal levels within 24 h. Caspase 3-mediated poly(ADP-ribose) polymerase cleavage is also inhibited in cells overexpressing hGSTA2-2. hGSTA2 transfection does not affect the function of antioxidant enzymes including GPx activity toward H 2 O 2 suggesting that the Alpha class GSTs play an important role in regulation of the intracellular concentrations of the lipid peroxidation products that may be involved in the signaling mechanisms of apoptosis.

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“…It is well established that mammalian GSTs play an important role in the detoxification of electrophilic xenobiotics (Mannervik and Danielson, 1988). Although endogenous substrates for mammalian GSTs have not been clearly defined, there is evidence that ␣-GSTs protect against oxidative stress by detoxifying reactive products generated by lipid peroxidation (Ålin et al, 1985;Ketterer and Coles, 1991;Singhal et al, 1992).In general, plant GSTs have not been as well characterized as mammalian GSTs. Plant cytosolic GSTs belong to the archaic class of GSTs (Meyer et al, 1991;Marrs, 1996).…”

mentioning

confidence: 99%

Isolation and Characterization of GlutathioneS-Transferase Isozymes from Sorghum1

1998

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Two glutathione S-transferase (GST) isozymes, A1/A1 and B1/B2, were purified from etiolated, O-1,3-dioxolan-2-yl-methyl-2,2,2,-trifluoro-4 -chloroacetophenone-oxime-treated sorghum (Sorghum bicolor L. Moench) shoots. GST A1/A1, a constitutively expressed homodimer, had a subunit molecular mass of 26 kD and an isoelectric point of 4.9. GST A1/A1 exhibited high activity with 1-chloro-2, 4,dinitrobenzene (CDNB) but low activity with the chloroacetanilide herbicide metolachlor. For GST A1/A1, the random, rapidequilibrium bireactant kinetic model provided a good description of the kinetic data for the substrates CDNB and glutathione (GSH). GST B1/B2 was a heterodimer with subunit molecular masses of 26 kD (designated the B1 subunit) and 28 kD (designated the B2 subunit) and a native isoelectric point of 4.8. GST B1/B2 exhibited low activity with CDNB and high activity with metolachlor as the substrate. The kinetics of GST B1/B2 activity with GSH and metolachlor fit a model describing a multisite enzyme having two binding sites with different affinities for these substrates. Both GST A1/A1 and GST B1/B2 exhibited GSH-conjugating activity with ethacrynic acid and GSH peroxidase activity with cumene hydroperoxide, 9-hydroperoxy-trans-10,cis-12-octadecadienoic acid and 13-hydroperoxy-cis-9,trans-11-octadecadienoic acid. Both GST A1/A1 and GST B1/B2 are glycoproteins, as indicated by their binding of concanavalin A. Polyclonal antibodies raised against GST A1/A1 exhibited cross-reactivity with the B1 subunit of GST B1/B2. Comparisons of the N-terminal amino acid sequences of the GST A1, B1, and B2 subunits with other type I -GSTs indicated a high degree of homology with the maize GST I subunit and a sugarcane GST.GSTs (EC 2.5.1.18) are dimeric enzymes found in mammals, insects, plants, and microbes that catalyze nucleophilic attack by the thiolate anion of GSH at electrophilic centers of hydrophobic molecules (Mannervik and Danielson, 1988). In addition to catalyzing GSH conjugation, GSTs also exhibit GSH peroxidase activity and ligandbinding functions (Mannervik and Danielson, 1988;Marrs, 1996). Mammalian GSTs compose a multigene family; in rat liver at least 13 different cytosolic GST subunits are found as either heterodimers or homodimers . Mammalian cytosolic GSTs have been divided into four classes (␣, , , and ) based on immunological, biochemical, and sequence similarities (Buetler and Eaton, 1992). It is well established that mammalian GSTs play an important role in the detoxification of electrophilic xenobiotics (Mannervik and Danielson, 1988). Although endogenous substrates for mammalian GSTs have not been clearly defined, there is evidence that ␣-GSTs protect against oxidative stress by detoxifying reactive products generated by lipid peroxidation (Ålin et al., 1985;Ketterer and Coles, 1991;Singhal et al., 1992).In general, plant GSTs have not been as well characterized as mammalian GSTs. Plant cytosolic GSTs belong to the archaic class of GSTs (Meyer et al., 1991;Marrs, 1996). This class, which is very heterog...

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Glutathione S-transferases of human lung: Characterization and evaluation of the protective role of the α-class isozymes against lipid peroxidation

Cited by 109 publications

References 40 publications

Microarray Expression Profiling Identifies Genes with Altered Expression in HDL-Deficient Mice

Microarray Expression Profiling Identifies Genes with Altered Expression in HDL-Deficient Mice

Role of Glutathione S-Transferases in Protection against Lipid Peroxidation

Isolation and Characterization of GlutathioneS-Transferase Isozymes from Sorghum1

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