• 농약과학회지
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• 제2권1호
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• pp.97-103
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• 1998

제초제 alachlor와 metolachlor가 처리된 수수의 생장에 대한 약해경감제 fluxofenim의 약해 경감효과와 경감기작의 하나로 추정되는 GST 활성에 대하여 조사하였다. 제초제 metolachlor와 alachlor는 수수(품종;G522DR)의 유묘생장을 크게 억제하였는데, 지상부 및 뿌리에 대한 50% 생장억제 농도가 각각 30.8, 28.8 ${\mu}M$과 4.48, 6.23 ${\mu}M$로 두 약제 모두 수수의 지상부에 대한 억제보다 뿌리에 대한 억제가 컸다. Fluxofenim을 종자에 처리하여 파종하고 metolachlor또는 alachlor을 처리하면 수수의 유묘생장이 회복되어 fluxofenim처리에 의한 약해경감 효과가 크게 나타났다. 약해경감제 fluxofenim을 처리한 것과 처리하지 않은 수수 유묘로부터 추출한 GST의 활성을 비교한 결과, fluxofenim을 처리한 수수의 유묘로부터 추출한 GST의 활성이 CDNB를 기질로 사용하였을때 70% 증가되었고, [$^{14}C$]-metolachlor을 기질로 사용하였을 때에도 82% 증가되었다. 따라서 약해경감제 fluxofenim을 처리한 수수와 처리하지 않은 수수의 metolachlor또는 alachlor에 대한 선택성의 차이는 fluxofenim 처리로 증가된 GST에 의한 metolachlor-glutathione 또는 alachlor-glutathione conjugation되기 때문인 것으로 생각된다.

• BMB Reports
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• 제38권3호
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• pp.300-306
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• 2005

Tamoxifen citrate is an anti-estrogenic drug used for the treatment of breast cancer. It showed a degree of hepatic carcinogenesis, when it used for long term as it can decrease the hexose monophosphate shunt and thereby increasing the incidence of oxidative stress in liver rat cells leading to liver injury. In this study, a model of liver injury in female rats was done by intraperitoneal injection of tamoxifen in a dose of 45 mg/kg body weight for 7 successive days. This model produced a state of oxidative stress accompanied with liver injury as noticed by significant declines in the antioxidant enzymes (glutathione-S-transferase, glutathione peroxidase and catalase) and reduced glutathione concomitant with significant elevations in TBARS (thiobarbituric acid reactive substance) and liver transaminases; sGPT (serum glutamate pyruvate transaminase) and sGOT (serum glutamate oxaloacetate transaminase) levels. The oral administration of dimethyl dimethoxy biphenyl dicarboxylate (DDB) in a dose of 200 mg/kg body weight daily for 10 successive days, resulted in alleviation of the oxidative stress status of tamoxifen-intoxicated liver injury in rats as observed by significant increments in the antioxidant enzymes (glutathione-S-transferase, glutathione peroxidase and catalase) and reduced glutathione concomitant with significant decrements in TBARS and liver transaminases; sGPT and sGOT levels. The administration of DDB before tamoxifen intoxication (as protection) is more little effective than its curative effect against tamoxifen-induced liver injury. The data obtained from this study speculated that DDB can mediate its biochemical effects through the enhancement of the antioxidant enzyme activities and reduced glutathione level as well as decreasing lipid peroxides.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.938-943
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• 2004

A gene responsible for the chlorothalonil-biotransformation was cloned from the chromosomal DNA of Ochrobactrum anthropi SH35B, an isolated bacterium strain from soil. We determined the nucleotide sequences and found an open reading frame for glutathione S-transferase (GST). The drug-hypersensitive Escherichia coli KAM3 cells transformed with a plasmid carrying the GST gene can grow in the presence of chlorothalonil. The GST of O. anthropi SH35B was expressed in E. coli and purified by affinity chromatography. The fungicide chlorothalonil was rapidly transformed by the purified GST in the presence of glutathione. No significant difference in the chlorothalonil-biotransformation effect was observed among the thiol compounds (cysteine, reduced glutathione, and $\beta$-mercaptoethanol). Thus, the result reported here is the first evidence on the chlorothalonil-biotransformation by conjugation with the cellular free thiol groups, especially glutathione, catalyzed by the bacterial GST.

• 한국환경농학회지
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• 제16권1호
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• pp.37-43
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• 1997

율무의 simazine에 대한 내성이 GSH와의 conjugation에 의한 약제의 불활성화 반응에 기인하는지 여부를 검정하기 위하여 화본과 작물인 옥수수와 비교하여 약제의 흡수 및 대사속도, GSH 함량 및 GST 활성, GST isozyme의 발현 양상 및 작용점에서의 약제에 대한 감수성의 차이를 조사하였다. 율무와 옥수수 모두 배양액중에 포함된 simazine에 대하여 약제처리 초기부터 빠른 흡수를 보였으며, 처리 12시간 후에는 두 초종 간의 흡수속도에 있어서 차이가 인정되지 않았다. 율무에 흡수된 simazine은 GSH-simazine conjugate의 형태로 신속히 대사되었으며, 특히 약제처리 $1{\sim}6$시간 까지의 처리 초기에는 율무에서의 대사속도가 옥수수에 비하여 약 2배정도 빠르게 나타났다. 두 초종 모두 초엽부에서 가장 높은 GSH 함량 분포를 보였다. 초종별로는 율무에서의 GSH 함량이 옥수수에 비하여 전반적으로 높게 나타났으며, 특히 율무의 초엽부와 근부에서 옥수수에 비하여 GSH함량이 각각 1.5배와 2.3배씩 높게 나타났다. GST[CDNB] 활성은 초종 간에 차이가 인정되지 않았으나, GST[simazine] 활성에 있어서는 옥수수에 비하여 율무의 모든 조직에서 약 2배 정도 높은 활성을 보였다. 두 초종 모두 지상부에서의 GST[simazine]활성이 근부에 비하여 $20{\sim}30%$ 정도 높게 나타났다. 율무와 옥수수에서 나타난 simazine에 대하여 특이성을 갖는 GST 활성의 차이를 이해하기 위하여 FPLC-anion exchange 컬럼을 이용하여 GST isozyme 활성을 조사하였다. 율무에 있어서 GST[CDNB] 활성을 보이는 1개 피크와 GST[simazine] 활성을 보이는 2개의 피크가 옥수수의 활성 피크와 같은 NaCl 농도범위에서 나타났으며, 이들의 활성은 옥수수에 비하여 높게 나타났다. 옥수수에서는 나타나지 않은 1개의 GST[simazine] 활성 피크가 율무에서 검출되었으며, 이 GST[simazine] 활성은 CDNB에 대하여 전혀 활성을 나타내지 않았기 때문에 율무에 simazine 특이성의 다른 GST isozyme이 존재함을 시사해 주고 있다. 율무와 옥수수의 완전 엽으로부터 추출한 엽록체 thylakoid막에 의한 전자전달 활성은 simazine $1{\sim}100\;{\mu}M$ 수준에서 매우 유사한 경향으로 억제되었다. 이상의 결과들은 율무의 simazine 내성이 흡수된 약제가 GSH와의 conjugation에 의하여 신속히 불활성화됨으로써 나타나며, 이러한 simazine 대사활성의 차이는 식물체에 내재하는 GSH 함량 및 GST 활성과 밀접한 상관성이 있음을 시사해 주고 있다.

• BMB Reports
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• 제31권1호
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• pp.77-82
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• 1998

To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal1 gene, encoding the ${\beta}$ subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene, the ${\alpha}$ subunit gene of GGPT-I. GGPT-I was highly expressed as a fusion protein with glutathione S-transferase (GST) in E. coli, purified to homogeneity by glutathione-agarose affinity chromatography, and the GST moiety was excised by thrombin treatment. The purified yeast GGPT-I showed a dose-dependent increase in the transferase activity, and its apparent $K_m$ value for an undecapeptide fused with GST (GST-PEP) was $0.66\;{\mu}M$ and the apparent value for geranylgeranyl pyrophosphate (GGPP) was $0.071\;{\mu}M$.

• 농약과학회지
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• 제6권1호
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• pp.31-35
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• 2002

본 논문의 목적은 benfuracarb 원제(90.2%)에 함유된 불순물의 glutathione-S-transferase와 amidase에 대한 저해 특성과 해당 불순물의 구조를 밝히는데 있다. Benfuracarb 원제, 유효성분 및 불순물은 glutathione-S-transferase(GST)를 효과적으로 저해하였으나, 그 저해력은 GST 효소 저해제인 ethacrynic acid의 저해력보다는 낮았다. 즉, GST에 대한 benfuracarb 원제, 유효성분 및 불순물의 $I_{50}$은 각각 $9.7{\times}10^{-4}M,\;>1.0{\times}10^{-3}M,\;1.8{\times}10^{-4}M$이었으나, ethacrynic acid의 $I_{50}$은 $1.7{\times}10^{-5}M$이었다. Benfuracarb 원제, 유효성분 및 불순물은 amidase를 저해하였는데, 이들의 효소저해력은 iprobenfos의 저해력($I_{50},\;8.2{\times}10^{-7}M$)보다는 낮은 $6.0{\times}10^{-5}M,\;4.3{\times}10^{-4}M,\;7.6{\times}10^{-5}M$이었다. Benfuracarb 원제에는 4종의 불순물(IM $1{\sim}4$)이 검출되었는데, 이들 중 IM 2와 3은 GST와 amidase의 활성을 저해하였던 반면, IM 4는 효소활성을 저해하지 않았다. 이들 불순물 중 효소활성 저해특성을 갖는 IM 2와 3을 IR, $^1H$-NMR, $^{13}C$-NMR, LC-MS를 이용하여 구조를 분석한 결과, IM 2는 ethyl-N-isopropylamino propionate로, 그리고 IM 3은 ethyl-N-isopropyl-N-(chlorosulfenyl) aminopropionate로 확인되었다.

• 대한한방내과학회지
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• 제25권1호
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• pp.81-91
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• 2004

Objectives : The aim of this study is to investigate the inhibitory effect of Chungganhaeju-Tang on alcohol induced human hepatic cell apoptosis by synthesis of glutathione. Methods : The amount of glutathione in HepG2 cell was measured with colorimetric glutathione assay kit and glutathione-conjugated CDNB(1-chloro-2,4-dinitrobenzene) at $37^{\circ}C$ and then measured by spectrometry to assess the activity of glutathione S-transferase. Results : The synthesis of glutathione and the activity of glutathione S-transferase in HepG2 cell were promoted by Chungganhaeju-Tang and increased in dose/time-dependent manner. Chungganhaeju-Tang inhibited apoptosis induced by ethanol and acetaldehyde dependent to treatment dosage. In Buthione sulfoximine, a glutathione synthesis inhibitor, treated case, the synthesis of glutathione was inhibited and in Chungganhaeju-Tang treated case, the synthesis of glutathione is promoted with or without Buthione sulfoximine. The present findings suggest that Chungganhaeju-Tang inhibits alcohol induced apoptosis by synthesis of glutathione in HepG2 cell. Conclusions : The result indicates that Chungganhaeju-Tang protects human hepatic cell by glutathione synthesis and made the liver recover from alcohol induced damage.

• BMB Reports
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• 제29권2호
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• pp.111-115
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• 1996

A novel glutathione S-transferase from Pseudomonas sp. DJ77 was expressed in E. coli and purified by glutathione-affinity chromatography. The enzyme was composed of two identical subunits. The molecular size of the enzyme was 42 kDa by sephadex G-150 gel permeation chromatography and Mr of each subunit was 23 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. pI value of the enzyme was approximately 5.8 by isoelectric focusing. This enzyme showed the highest activity toward 1-chloro-2,4-dinitrobenzene as the electrophilic substrate. The relative activities toward p-nitrobenzyl chloride and 1,2-dichloro-4-nitrobenzene were 3.8% and 1.3% of the activity toward 1-chloro-2,4-dinitrobenzene, respectively. $K_m$ and $V_{max}$ values for 1-chloro-2,4-dinitrobenzene calculated by Lineweaver-Burk plot were 0.76 mM and $14.81\;{\mu}mol/min/mg$, respectively, and those for glutathione were 6.23 mM and $64.93\;{\mu}mol/min/mg$, respectively. The enzyme showed highest glutathione S-transferase activity at pH 8.0 and was stable between pH 6.0 and 9.0. The enzyme retained its activity up to $35^{\circ}C$ for 90 min but was unstable above $45^{\circ}C$.

• 동아시아식생활학회지
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• 제6권1호
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• pp.41-49
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• 1996

This research was performed to Investigate the effects of green tea on the lipid composition of serum and liver and the specific activities of antioxidative enzymes. Male Sprague Dawley rats were fed 10% fat diet with lard and fish oil. Powdered green tea was added to the lard and fish oil diet at the level of 0.1% and 1%. After 6 weeks of feeding, serum and liver were obtained from experimental rats. Then we measured the concentration of total cholesterol, HDL-cholesterol and triglyceride. From liver cytosolic fraction, we analized the specific activities of superoxide dismutase, glutathione peroxidase and glutathione S-transferase. The level of total cholesterol and triglyceride were decreased and the ratio of HDL-cholesterol to total cholesterol was increased by the fish oil in the serum. But in the liver, the level of total cholesterol was increased by the fish oil and green tea than the lard. The specific activities of glutathione S-transferase were more increased in the fish oil than the lard. There was not effect of the green tea of daily dose on the lipid composition of serum and liver and the specific activities of antioxidative enzymes in rats.

• BMB Reports
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• 제30권4호
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• pp.296-298
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• 1997

A bacterial glutathione S-transferase from Pseudomonas sp. DJ77 has been crystallized. The crystals diffract to at least $2.3\;\AA$ resolution, and belong to the orthorhombic space group $P2_{1}2_{1}2_{1}$, with cell parameters $a=97.4\;\AA,\;b=100.3\;\AA$, and $c=46.0\;\AA$. There is one dimer molecule of pGST per crystallographic asymmetric unit. with the crystal volume per protein mass of $2.34\;\AA^3/dalton$ and a solvent content of about 47% (v/v).