• Journal of Life Science
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• 제9권1호
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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.

• Bulletin of the Korean Chemical Society
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• 제33권12호
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• pp.4169-4172
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• 2012

To elucidate the roles of cysteine residues in rice Phi-class GST F3, in this study, all three cysteine residues were replaced with alanine by site-directed mutagenesis in order to obtain mutants C22A, C73A and C77A. Three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitutions of Cys73 and Cys77 residues in OsGSTF3 with alanine did not affect the glutathione conjugation activities, showing non-essentiality of these residues. On the other hand, the substitution of Cys22 residue with alanine resulted in approximately a 60% loss of specific activity toward ethacrynic acid. Moreover, the ${K_m}^{CDNB}$ value of the mutant C22A was approximately 2.2 fold larger than that of the wild type. From these results, the evolutionally conserved cysteine 22 residue seems to participate rather in the structural stability of the active site in OsGSTF3 by stabilizing the electrophilic substrates-binding site's conformation than in the substrate binding directly.

• 한국환경농학회지
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• 제8권1호
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• pp.47-54
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• 1989

동양종(東洋種)꿀벌 (Apis cerana F.)에 대(對)한 살충제(殺蟲劑)의 독성(毒性) 및 해독능력(解毒能力)을 조사(調査)하고 농약한계 사용량 결정에 기여하기 위하여 7가지 대표적인 살충제의 꿀벌에 대한 독성 및 해독효소의 활성을 조사하였다. 효소 활성은 해독효소로 알려진 microsomal oxidases, glutathione S-transferasecs, esterase와 DDT-dehydrochlorinase를 조사했고 성충(成蟲)일벌의 중장(中腸)을 사용하여 측정하였다. $LC_{50}$치의 측정 결과는 다음과 같다. 1. 공시 살충제중 DDT가 19ppm으로 독성(毒性)이 가장 낮았고 EPN이 0.75ppm으로 독성(毒性)이 가장 강(强)했다. 2. 준치사농도(準致死濃度)의 농약(農藥)이 성충(成蟲)일벌의 microsomal oxidase에 미치는 영향은 malathion 및 demeton S-methyl 처리가 aldrin epoxidase활성을 저해시켰고 N-demethylase활성은 carbayl 처리구에서 증대(增大)되었다. 3. Glutathione S-transferase(DCNB conjugation)활성은 diazinon과 malathion처리구에서 증대되었다. 4. Esterase는 malathion 및 permethrin처리구에서 ${\alpha}-NA$ esterase 활성(活性)의 저해(沮害)를 보였고 carboxylesterase와 AchE 활성은 거의 영향이 없었다. 5. DDT-dehydrochlorinase 활성은 carbaryl, malathion과 demeton S-methyl 처리구에서 저해를 보였다.

• 대한한의학방제학회지
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• 제9권1호
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• pp.355-366
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• 2001

Objectives : Oldenlandiae Diffusae herba has been used as a natural drug for tumor, inflammation and liver disease in traditional medicine. This study was performed in order to investigate the antioxidative effects of Oldenlandiae Diffusae herba methanol extract(ODHM) on acetaminophen induced acute liver injury in mice. Methods : In order to investigate the protective effect of ODHM on acute hepatic injury in vivo, ICR mice were pretreated with ODHM, and then treated with acetaminophen(500mg/kg). And the levels of LPO and glutathione(GSH), antioxidative enzyme activities were measured. The levels of LPO were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) was assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. And Total SH and GSH levels were measured. Results : In vivo study, LPO levels of acetaminophen treatment group were significantly higher than other groups. This increased level was significantly reduced by ODHM pretreatment. The acetaminophen treatment resulted in a decrease of catalase, GPX, SOD and GST activities. By contrast, ODHM pretreatment markedly increased compare to those of untreated groups. Total SH and GSH levels were reduced by of acetaminophen treatment, and ODHM pretreatment significantly increased GSH levels.

• Toxicological Research
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• 제11권2호
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• pp.295-302
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• 1995

Menadione-ghitathione conjugate (MEN-SG), a metabolite of menadione, is known to be a redoxcycler in rat hepatocyte subcellular fraction. Therefore, it was assumed that MEN-SG could exert cytotoxlclty to ral platelets, another target tissue of menadione. We first synthesized MEN-SG, the identity of which was verified by mass, $^1{H}$-NMR and UV-visible spectra. In addition, the stability of MEN-SG was investigated in biological assay system. MEN-SG was degraded in a time-dependent manner in DMSO which had been used as a vehicle and thus, tris-HCl buffer was used as a vehicle of MEN-SG despite the low solubility in it. Perchloric acid as well as platelets itself did not affect the stability of MEN-SG. Our next attempt was the evaluation of cytotoxicity of MEN-SG in rat platelets. MEN-SG did not induce cytotoxicity to rat platelets measured by two different methods, LDH release and turbidity changes. The extents of oxygen consumption by MEN-SG in intact platelets were significantly lower than those by menadione, though it had been observed that oxygen consumptions by menadione and MENSG were similar in subcellular fractioas of platelets. These results suggest that MEN-SG is not toxic to rat platelets despite its redox cycling capacity and glutathione conjugation reaction of menadione could be regarded as a detoxification process.

• Journal of Ginseng Research
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• 제36권4호
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• pp.449-460
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• 2012

Plants have versatile detoxification systems to encounter the phytotoxicity of the wide range of natural and synthetic compounds present in the environment. Glutathione S-transferase (GST) is an enzyme that detoxifies natural and exogenous toxic compounds by conjugation with glutathione (GSH). Recently, several roles of GST giving stress tolerance in plants have demonstrated, but little is known about the role of ginseng GSTs. Therefore, this work aimed to provide further information on the GST gene present in Panax ginseng genome as well as its expression and function. A GST cDNA (PgGST) was isolated from P. ginseng cDNA library, and it showed the amino acid sequence similarity with theta type of GSTs. PgGST in ginseng plant was induced by exposure to metals, plant hormone, heavy metals, and high light irradiance. To improve the resistance against environmental stresses, full-length cDNA of PgGST was introduced into Nicotiana tabacum. Overexpression of PgGST led to twofold increase in GST-specific activity compared to the non-transgenic plants, and the GST overexpressed plant showed resistance against herbicide phosphinothricin. The results suggested that the PgGST isolated from ginseng might have a role in the protection mechanism against toxic materials such as heavy metals and herbicides.

• Fisheries and Aquatic Sciences
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• 제27권5호
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• pp.314-328
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• 2024

The mechanism for the elimination of xenobiotics undergoes three different phases of reactions in organisms. Among these, glutathione S-transferases (GSTs) are classified as phase II detoxification enzymes, catalyzing the conjugation of electrophilic substrates to glutathione or reduced hydroperoxides. This study aimed to investigate the molecular characteristics, detoxification functions, and immune responses of GST omega (LhGSTO1) and kappa (LhGSTK1) in redlip mullet. The open reading frames of LhGSTO1 (720 bp) and LhGSTK1 (687 bp) encoded proteins of 239 and 228 amino acids, respectively. Sequence analysis revealed that LhGSTO1 and LhGSTK1 possessed GSH-binding sites in their N-terminal domains. Substrate-binding sites in the C-terminal domain were exclusively identified in LhGSTO1. In the tissue-specific transcription profile analysis, both LhGSTO1 and LhGSTK1 were ubiquitously expressed in all tissues of healthy mullets. Temporal expression analysis of LhGSTO1 and LhGSTK1 in the blood showed that their expression was significantly modulated by polyinosinic:polycytidylic (poly I:C), lipopolysaccharide (LPS), and Lactococcus garvieae. Different chemical and cellular assays were performed to assess the detoxification and cellular protective abilities of the two proteins. A substrate specificity test using the recombinant proteins revealed that both proteins possessed specific activity toward 1-chloro-2,4-dinitrobenzene (CDNB). In the disk diffusion assay, the smallest clearance zones were observed for LhGSTO1 and LGSTK1 against CdCl₂. In the cell protection assay, both LhGSTO1 and LhGSTK1 showed significant Cd detoxification ability compared to the control. Collectively, these results demonstrate that GST omega and kappa are involved in host defense against immune stimulants and xenobiotics in redlip mullet.

• Applied Biological Chemistry
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• 제31권3호
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• pp.241-248
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• 1988

서양종(西洋種) 꿀벌(Apis mellifera L)에 대(對)한 살충제의 독성(毒性) 및 해독능력(解毒能力)을 조사(調査)하고 농약한계사용량(農藥限界使用量) 결정(決定)에 기여(寄與)하기 위하여 7가지 대표적(代表的)인 살충제의 꿀벌에 대한 독성(毒性)및 해독효소(解毒酵素)의 활성(活性)을 조사(調査)하였다. 효소활성(酵素活性)은 해독효소(解毒酵素)로 알려진 microsomal oxidases, glutathione S-transferases, esterases 와 DDT-dehydrochlorinase를 조사(調査)했고 성충(成？)일별의 중장(中腸)을 사용(使用)하여 측정(測定)하였다. $LC_{50}$치(値)의 측정결과(測定決果) 공시(供試) 살충제중(殺？劑中) DDT가 58ppm으로 독성(毒性)이 가장 낮았고 EPN이 1.61ppm으로 독성(毒性)이 가장 강(强)했다. 준치사농도(準致死濃度)의 농약(農藥)이 성충(成？)일벌의 microsomal oxidase에 미치는 영향은 malathion 및 permethrin 처리(處理)가 aldrin epoxidase 활성(活性)을 저해(沮害)시켰고 N-demethylase 활성(活性)은 diazinon 처리구(處理區)에서 증대(增大)되었다. Ghtathione S-transferarse(DCNB conjugation) 활성(活性)은 diazinon과 permethrin 처리구(處理區)에서 증대(增大)되었다. Esterase는 malathion 및 permethrin 처리구(處理區)에서 ${\alpha}-NA$ esterase활성(活性)의 저해(沮害)를 보였고 diazinon처리구(處理區)에서 carboxylesterase활성(活性)이 증대(增大)되었으며 AChE 활성(活性)은 거의 영향(影響)이 없었다. DDT-dehy drochlorinase활성(活性)은 carbaryl 및 permethrin 처리구(處理區)에서 증대(增大)되었다.

• Bulletin of the Korean Chemical Society
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• 제26권3호
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• pp.433-439
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• 2005

To gain further insight on the relationship between structure and functions of glutathione S-transferase (GST), the three tyrosine 108 mutants, Y108A, Y108F, and Y108W, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitution of Tyr 108 with alanine resulted in significant decrease of the GSH-conjugation activity and the GSH peroxidase activity, but approximately 63% increase of steroid isomerase activity toward ${\Delta}^5$–[androstene 3,17-dione. On the other hand, the substitution of Tyr 108 with phenylalanine resulted in decreases of $k_{cat}\;and\;k_{cat}/K_m{^{EPNP}}$ by 2 orders of magnitude, suggesting that Tyr 108 residue of hGSTP1-1 are considered to be important for the catalysis and the binding of the epoxide substrates. The substitution of Tyr 108 with tryptophan resulted in significant decreases of the specific activities toward EPNP, cumene hydroperoxide and ${\Delta}^5$–ndrostene 3,17-dione, but approximately 2-fold increase on the enzyme-catalyzed addition of GSH to DCNB. We conclude from these results that Tyr 108 in hGST P1-1 plays very different roles depending upon the nature of the electrophilic substrates.

• Archives of Pharmacal Research
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• 제29권2호
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• pp.172-177
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• 2006

Hepatotoxic potential of 2, 3-dibromopropene (2, 3-DBPE) and its conjugation with glutathione (GSH) were investigated in male ICR mice. Treatment of mice with 20, 50, and 100 mg/kg of 2, 3-DBPE for 24 h caused elevation of serum alanine aminotransferase and aspartate aminotransferase activities. The hepatic content of GSH was not changed by 2, 3-DBPE. Meanwhile, the GSH content was slightly reduced when mice were treated with 2, 3-DBPE for 6 h and significantly increased 12 h after the treatment. Subsequently, a possible formation of GSH conjugate of 2, 3-DBPE was investigated in vivo. After the animals were treated orally with 20, 50, and 100 mg/kg of 2, 3-DBPE, the animals were subjected to necropsy 6, 12, and 24 h later. A conjugate of S-2-bromopropenyl GSH was identified in liver and serum treated with 100 mg/kg of 2, 3-DBPE by using liquid chromatography-electrospray ionization tandem mass spectrometry. The protonated molecular ions $[M+H]^+$ of S-2-bromopropenyl GSH were observed at m/z 425.9 and 428.1 in the positive ESI spectrum with a retention time of 6.35 and 6.39 min, respectively. In a time-course study in livers following an oral treatment of mice with 100 mg/kg of 2, 3-DBPE for 6, 12, and 24 h, the 2, 3-DBPE GSH conjugate was detected maximally 6 h after the treatment. The present results suggested that 2, 3-DBPE-induced hepatotoxicity might be related with the production of its GSH conjugate.