• Environmental Mutagens and Carcinogens
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• v.24 no.1
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• pp.19-24
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• 2004

In order to understand the mechanism of the regulation of drug metabolizing enzyme gene expression, we have studied the induction of CYP1A1 and $GST\alpha,$ $\mu,$ $\pi$ enzymes in Japanese monkey and rhesus monkey after the treatment with 3-methylcholanthrene (3MC) and di-n-butyl phthalate (DBP) and bisphenol A (BPA). The levels of mRNA were measured by RT-PCR in brain, intestine and liver. In the case of adult monkey, treatment with 3MC induced CYP1A1 mRNA in intestine by 11-fold. The treatment with DBP induced CYP1A1 mRNA. Effects of 3MC and DBP on GST mRNA expression was not clear. But $GST\mu$ was slightly inhibited by the treatment with 3MC and DBP. $GST\alpha$ was induced in intestine by 1.5-fold. $GST\pi$ was slightly induced by the treatment with 3MC and DBP in intestine. In the case of fetus monkey, the basal levels of fetus CYP1A1 mRNA and GSTs mRNA were relatively low compared to adult monkey. As the age of monkey increased, the basal levels of CYP1A1 mRNA were also increased. 3MC induced the expression of CYP1A1 mRNA didn't significantly induce CYP1A1 mRNA in intestine. The levels of $GST\mu$ and $GST\pi$ were not changed by the treatment with 3MC and DBP. $GST\pi$ was slightly induced by the treatment with 3MC and DBP.

• Journal of Bio-Environment Control
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• v.11 no.3
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• pp.139-143
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• 2002

Cotton Glutathione S-Transferase (GST: EC 2.5.1.18) was cloned and overexpressed in tobacco (Nicotiana tabacum) plants. Northern blot analysis confirmed the successful transformation of cotton gst gene in tobacco plant. Type I and Type ll transcript patterns were identified in transgenic tobacco plants and only Type I transcripts were discussed in this paper, The activity of GST in the type II transgenic plants was about 1.5-fold higher than those of the wild type and non-expresser by using 1-chloro-2,4-dinitrobenzene (CDNB) and reduced glutathione as the substrate. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type II transgenic plants produced functional GST in the cells. The effects of cotton GST in the seedlings was evaluated by growing the control and transgenic seedlings at $15^{\circ}C$ in the growth chamber in the light. Overexpressors were grown well compared to the control plants (non-expressors). lo test far tolerance to salinity, seeds of Gh-5 overexpressors and the wild type Xanthi seedlings were grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings on 50 and 100 mM NaCl solution. There was no difference in growth rate at 150 and 200mM NaCl concentration.

• Environmental Mutagens and Carcinogens
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• v.24 no.2
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• pp.73-78
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• 2004

In order to understand the mechanism of the regulation of drug metabolizing enzyme gene expression, we have studied the induction of CYP1A1 and GST$\alpha$, $\mu$, $\pi$ enzymes in Japanese monkey and rhesus monkey after the treatment with 3-methylcholanthrene (3MC) and di-n- butyl phthalate (DBP) and bisphenol A (BPA). The levels of mRNA were measured by RT-PCR in brain, intestine and liver. In the case of adult monkey, treatment with 3MC induced CYP1A1 mRNA in liver by 10-fold. The treatment with DBP induced CYP1A1 mRNA. Effects of 3MC and DBP on GST mRNA expression was not clear. But GST$\mu$ was slightly inhibited by the treatment with 3MC and DBP. GST$\pi$ was not induced by the treatment with 3MC and DBP in liver. GST$\alpha$ was slightly induced by the treatment with 3MC and DBP in liver. In the case of fetus monkey, the basal levels of fetus CYP1A1 mRNA and GSTs mRNA were relatively low compared to adult monkey. As the age of monkey increased, the basal levels of CYP1A1 mRNA were also increased. 3MC induced the expression of CYP1A1 mRNA in liver. The levels of GST$\mu$ and GST$\alpha$ were not changed by the treatment with 3MC and DBP. GST$\pi$ was slightly induced by the treatment with 3MC and DBP.

• Sleep Medicine and Psychophysiology
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• v.22 no.1
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• pp.25-29
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• 2015

Objectives: The pathophysiology of restless legs syndrome (RLS) has not been fully elucidated. Oxidative stress might play a role in the development of RLS and other antipsychotic-induced side effects such as tardive dyskinesia. In the present study, we investigated whether the glutathione S-transferase (GST) gene polymorphisms are associated with antipsychotic-induced RLS in schizophrenia. Methods: We assessed antipsychotic-induced RLS symptoms in 190 Korean schizophrenic patients using the diagnostic criteria of the International Restless Legs Syndrome Study Group. The GST-M1, GST-T1 and GST-P1 loci were analyzed using PCR-based methods. Results: We divided the subjects into 2 groups: those with RLS symptoms (n = 96) and those without RLS symptoms (n = 94). There were no significant differences in the distributions of the GST-M1 genotypes (${\chi}^2=3.56$, p = 0.059), GST-T1 (${\chi}^2=0.51$, p = 0.476) and GST-P1 (${\chi}^2=0.57$, p = 0.821) between the 2 groups. Comparison of the RLS score among genotypes of the GST-M1 (t = -1.54, p = 0.125), GST-T1 (t = -0.02, p = 0.985) and GST-P1 (F = 0.58, p = 0.560) revealed no significant difference. Conclusion: These data suggest that GST gene polymorphisms do not confer increased susceptibility to RLS symptoms in schizophrenic patients. Future studies are necessary to evaluate the possible influences of other candidate genes involved in the reactive oxygen species system.

• The Korean Journal of Malacology
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• v.30 no.4
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• pp.399-408
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• 2014

Glutathione S-transferases (GSTs) are a superfamily of detoxification enzymes that primarily catalyze the nucleophilic addition of reduced glutathione to both endogenous and exogenous electrophiles. In this study, we isolated and characterized a full-length of alpha class GST cDNA from the abalone (Haliotis discus hannai). The abalone GST cDNA encodes a 223-amino acid polypeptide with a calculated molecular mass of 25.8 kDa and isoelectric point of 5.69. Multiple alignments and phylogenetic analysis with the deduced abalone GST protein revealed that it belongs to the alpha class GSTs and showed strong homology with disk abalone (Haliotis discus discus) putative alpha class GST. Abalone GST mRNA was ubiquitously detected in all tested tissues. GST mRNA expression was comparatively high in the mantle, gill, liver, and digestive duct, however, lowest in the hemocytes. Expression level of abalone GST mRNA in the mantle, gill, liver, and digestive duct was 182.7-fold, 114.8-fold, 4675.8-fold, 406.1-fold higher than in the hemocytes, respectively. Expression level of abalone GST mRNA in the liver was peaked at 6 h post-infection with Vibrio parahemolyticus and decreased at 12 h post-infection. While the expression level of abalone GST mRNA in the hemocytes was drastically increased at 3 h post-infection with Vibrio parahemolyticus. These results suggest that abalone GST is conserved through evolution and may play roles similar to its mammalian counterparts.

• BMB Reports
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• v.31 no.3
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• pp.287-295
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• 1998

Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of L-Ieucine, L-isoleucine, and L-valine. The sulfonylurea-resistant ALS gene from Nicotiana tabacum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS3 was used to transform Escherichia coli strain XL1-Blue, and the mutant tobacco ALS (mALS) was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). The fusion product GST-mALS was purified in a single step on a glutathione-Sepharose column. ALS activities of 0.9-2.5 ${\mu}mol/min/mg$ protein were observed in the GST-mALS, and the Km values for pyruvate, FAD, and TPP were 10.8-24.1, $(1.9-8.9){\times}10^{-3}$, and 0.14-0.38 mM, respectively. The purified GST-mALS was resistant to both the sulfonylurea and the triazolopyrimidine herbicides, and lost its sensitivity to end products, L-valine and L-leucine. For comparision, the tobacco wild-type recombinant ALS fused with GST, GST-wALS, was also characterized with respect to its pyruvate and cofactor bindings. These results suggest that the purified mutant recombinant tobacco ALS was functionally active, that the mutations resulting in herbicide resistance has affected pyruvate and cofactor bindings," and that the two classes of herbicides interact at a common site on the plant ALS.

• Journal of Life Science
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• v.9 no.1
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• pp.40-44
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• 1999

Glutathione S-transferase (GST) is a multifunctional protein that catalyzes the catalyzes the conjugation of glutathione with electrophilic compounds. It exists in a variety of isoenzy-matic froms with a wide range of substrate specificity and plays a pivotal role in detoxification of various drugs. In order to elucidate the GST-${\pi}$'s involvement of multidrug resistance (MDR) in drug-resistant tumor cell lines, we determined GST-${\pi}$ content by "1 step sandwich method". Consequently, adriamycin resistant cells of MCF-7 (MCF-7/ADM) have 7-fold increase of GST-${\pi}$ content than that of MCF-7 cells, while its {TEX}$IC_{50}${/TEX} was 116-fold greater than parent cell line. By northrn blotting, we compared whether MCF-7/ADM cells express GST-${\pi}$ mRNA. The GST-${\pi}$ mRNA expression in these cells was not inducible, but constitutive when treated for 24 h with a concentration of 0, 20, 200, and 2000 nM of adriamycin, respectively. Taken together, these results suggest that GST-${\pi}$ may not be directly associated with multidrug resistance in these human cancer cell lines.ell lines.

• Journal of Plant Biotechnology
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• v.4 no.3
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• pp.117-122
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• 2002

Cotton Glutathione S-Transferase(GST: EC 2.5.1.18) was cloned and Gh-5 cDNA was overexpressed in tobacco (Nicotiana tabacum) plants. The transformation of cotton GST in tobacco plant was confirmed by northern blot analysis. Type I and Type II transcript patterns were identified in Gh-5 transgenic tobacco plants. Type I transcripts was only discussed in this paper. Glutathione and 1-chloro-2,4-dinitrobenzene (CDNB) were used as the substrates, and the activity of GST in the type I transgenic plants was about 2.5-fold higher than the non-expressers and wild type tobacco plants. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type I transgenic plants produced functional GST in the cells. Type I showed higher GST specific activity than Type II in the transgenic plants. Control and transgenic seedlings were grown in the growth chamber and under the light at 15$^{\circ}C$, and the effects of cotton GST in the seedlings was evaluated. The growth rate of Gh-5 overexpressors was better than the control and non-transgenic tobacco plants. Salinity tolerance was also analyzed on the seeds of transgenic plants. Seeds of Gh-5 overexpressors and the wild type tobacco seedlings were germinated and grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings at both 50 and 100 mM NaCl solution. But at 0, 150, and 200 mM NaCl concentration, the difference in growth rate was not detected.

• Journal of Haehwa Medicine
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• v.14 no.2
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• pp.45-54
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• 2005

GST, an extract from 16 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present study was done to investigate whether GST has suppressive effects on activation of fibroblast-like sinoviocytes isolated from an RA patient. In tumor necrosis factor-a(TNF-a)/interleukin-1b(IL-1b) treated human sinoviocytes, The mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GST($100\;{\mu}g/ml$) suppressed the expression of proinflammatory cytokines and chemokines such as TNF-a, IL-1b, IL-6 and IL-8 compared with the control. The mRNA level of intracellular adhesion molecule-1(ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GST. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). In addition, GST considerably increased the mRNA levels of tissue inhibitors of matrix metalloproteinase-1(TIMP-1), while those of matrix metalloproteinase-3(MMP-3) were decreased. Taken together, these data suggested that GST might suppress the activation of sinoviocytes in hRA.

• BMB Reports
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• v.33 no.2
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• pp.179-183
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• 2000

The glutathione S-transferase (GST) activities of S-type and L-type thioltransferases (TTases), which are purified from the seeds and leaves of Arabidopsis thaliana, respectively, were identified and compared. The S-type and L-type TTases showed $K_m$ values of 9.72 mM and 3.18mM on 1-chloro-2,4-dinitrobenzene (CDNB), respectively, indicating the L-type TTase has higher affinity for CDNB. The GST activity of the L-type TTase was rapidly inactivated after being heated at $70^{\circ}C$ or higher. The GST activity of the S-type TTase remains active in a range of $30-90^{\circ}C$. $Hg^{2+}$ inhibited the GST activity of the S-type TTase, whereas $Ca^{2+}$ and $Cd^{2+}$ inhibited the GST activity of the L-type TTase. Our results suggest that the GST activities of two TTases of Arabidopsis thaliana may have different catalytic mechanisms. The importance of the co-existence of TTAse and GST activities in one protein remains to be elucidated.