• Title, Summary, Keyword: Glutathion-S-transferase

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Effect of Timing of IPTG Addition on Expression of Turnip Mosaic Virus Coat Protein Gene in Escherichia Coli (IPTG의 첨가 시간이 대장균(Escherichia coli)에서 순무 모자이크 바이러스(TuMV)의 외피단백질 발현에 미치는 영향)

  • Kim, Su-Joong;Park, Won-Mok;Ryu, Ki-Hyun;Lee, Sang-Seon;Lee, Se-Yong
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.248-254
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    • 1997
  • Expression vector (pGEX-Tu) for the coat protein (CP) gene of turnip mosaic virus Ca strain (TuMV-Ca) was constructed by incorporation of TuMV CP gene into pGEX-KG vector which had ${\beta}$-galactosidase gene and IPTG (isopropylthio-${\beta}$-D-galactoside) induction site. The results of ELISA and western hybridization indicated that optimal condition of the expression were when IPTG and western hybridization indicated that optimal condition of the expression were when IPTG induction was carried out on YTA medium with ampicillin in 2 hours after the E. coli seed inoculation ($A_{595}$=0.1/ml). TuMV CP gene was expressed with GST (Glutathion S-Transferase) gene fusion system, and the size of fusion protein was estimated to be 59kDa, for TuMV CP was 33 kDa and GST was 26 kDa.

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The Effect of Ginseng on the Hepatic Glutathione S-Transferase Activity (인삼이 간 Glutathione S-Transferase 활성에 미치는 효과)

  • 김낙두;김승희;김신근
    • YAKHAK HOEJI
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    • v.25 no.4
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    • pp.153-160
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    • 1981
  • The investigation aimed to study the effect of ginseng on the hepatic glutathion S-transferase activity. The ginseng methanol extract was administered to rats and mice for 10 days and their hepatic gluthatione S-transferase activities were measured by the method of Habig et al. Glutathione S-transferase activities in the rat treated with 100 and 500mg/kg ginseng methanol extract were increased by 13.4% and 17.10%, respectively and their increases were statistically significant. Similar results were also found in the mouse treated with ginseng 100mg/kg methanol extract. To investigate components of the extract which induce the enzyme, the methanol extract was fractionated into ether and butanol fraction and their effect on the enzyme was compared. Glutathione S-transferase activities in the rat treated with ether fraction were increased by 13.1%, similar to that obtained with ginseng methanol extract, whereas, butanol fraction did not show any increase in the enzyme activities. In the rats treated with maltol, one of the components in ether fraction, 5mg/kg for 10 days, activity of glutathione S-transferase was increased by 7.89%, but its increase was not significantly different from control group. Therefore, it may be concluded that ginseng methanol extract and its ether soluble fraction had effect on the elevation of glutathione S-transferase activities, whereas, butanol fraction of ginseng methanol extract had no effect on the enzyme.

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Induction of Quinone Reductase and Glutathion S-transferase in Hepatoma Cells by Citrus aurantium Linn (Jikak) Produced in Cheju Island (지각 추출물이 quinone reductase 및 glutathion s-transferase의 유도활성에 미치는 영향)

  • Yu, Mi-Hee;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.261-267
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    • 2005
  • Phase II enzymes are transcriptionally induced by synthetic chemical agents and natural products, and such induction plays critical roles in protection against chemical carcinogens and other toxic xenobiotics. To discover natural products for use as cancer chemopreventive agents, the ability of Citrus aurantium Linn (Jikak) to induce activities of quinone reductase (QR) and glutathione S-transferase (GST) in wild-type murine hepatoma cell line (Hepa 1c1c7) and Ah-receptor-defective mutant of the same cell line (Bprcl) was investigated. Hexane and chloroform fractions of C. aurantium Linn (Jikak) at doses not exhibiting cytotoxicity were effective inducers of QR (${\sim}1.8-fold$) and GST (${\sim}1.5-fold$) in Hepa 1c1c7 cells, whereas showed low QR induction potency in Bprcl cells, which indicates they have weak monofunctional action. Results suggest C. aurantium Linn (Jikak) as potentially useful cancer chemopteventive agent.

Effect of Cyclohexane Treatment on Serum Level of Glutathione S-Transferase Activity in Liver Damaged Rats ($CCl_4$ 에 의한 간손상 모델 실험동물에 있어서 cyclohexane 투여가 혈청 glutathione S-transferase 활성에 미치는 영향)

  • 오정대;윤종국
    • Journal of Environmental Health Sciences
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    • v.29 no.2
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    • pp.80-86
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    • 2003
  • To evaluate the effect of cyclohexane(CH) treatment on the serum levels of glutathion S-transferase(GST) activity in liver damaged animals, damaged liver was induced with pretreatment of 50% $CCl_4$ dissolved in olive oil (0.1 m1/100g body weight) intraperitoneally 17 times every other day. To $CCl_4$-treated rats, CH (1.56 g/kg body weight, i.p) was injected once and then the animals were sacrificed at 4 hours after injection of CH. The $CCl_4$-treated animals were identified as severe liver damage on the basis of liver functional findings, 1,e, increased serum levels of alanine aminotransferase(ALT), alkaline phosphate(ALP) and xanthine oxidase(XO) activities. On the other hand, $CCl_4$-treated animals injected with CH once($CCl_4$-pretreated animals) showed more decreased serum levels of ALT and XO, and more increased those of ALP rather than $CCl_4$-treated animals. In case of comparing the GST with ALT activity in liver, both $CCl_4$-treated and pretreated animals showed similar changing pattern of enzyme actvity. Especially $CCl_4$-pretreated animals showed significantly increased serum level of GST actvity compared with the $CCl_4$-treated those, whereas those of ALT showed reversed tendency. In aspects of GST enzyme kinetics, $CCl_4$-pretreated animals showed higher Vmax of liver GST enzyme than $CCl_4$-treated animals. In conclusion, injection of CH to the liver damaged rats led to enhanced liver damage and more increased activity of serum GST which may be chiefly caused by the enzyme induction.

Protective Effects of Angelica tenuissima Nakai on Hepatotoxicity by Carbon Tetrachloride in Rats (사염화탄소에 의한 간손상에 미치는 고본의 보호작용)

  • 정춘식;정기화
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.211-217
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    • 2002
  • Hepatoprotective activity of methanol extract of Angelica tenuissima Nakai on the $CCl_4$-induced hepatotoxicity was investigated. To elucidate the hepatoprotective activity and free radical scavenging effect, we examined alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, total protein, cholesterol, malondialdehyde (MDA) levels in serum and activities of superoxide dismutase (SOD), catalase (CAT) in hepatic tissue as compared with those of carbon tetrachloride-induced rats. The action mechanism also has been estimated by quantative analysis of cytochrome P450 (CYP), NADPH-CYP reductase for phase I metabolism and glutathion (GSH), glutathion S-transferase (GST) level for phase II metabolsim. Treatment of Angelica tenuissima methanol extract significantly lowered the levels of alanine aminotransferase and aspartate aminotransferase. In addition, the levels of cholesterol, triglyceride, MDA, CAT were decreased, and SOD was activated. This result indicates that the hepatoprotective effect of Angelica tenuissima methanol extract on the CCl4-induced hepatotoxicity would be originated from reduction of the NADPH-CYP reductase, GSH and the enhancement of the activities of GST, CYP.

Studies on the Antimutagenic Effect of Acanthopanax sessiliflorum Components (오가피 성분의 항돌연변이원성에 관한 연구)

  • Chung, Kyu-Charn;Baek, Suk-Hwan;Nam, Kyung-Soo
    • YAKHAK HOEJI
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    • v.32 no.1
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    • pp.14-19
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    • 1988
  • Effects of acanthopanax cortex extracts on glutathion S-transferase (GST), glutathion peroxidase (GSH-px) and superoxide dismutase (SOD) activities related to 7,12-dimethyl-benz(a)anthracene(DMBA) metabolism and on DMBA-induced mutagenicity were investigated in this study. From the comparative study of three extracts, it was found that butanol extract was more potent than other extracts in increment of GST, GSH-px and SOD activities and in inhibitory effects of lipoperoxide formation of liver. Also ether and butanol extracts inhibited DMBA-induced mutagenicity, showing 33% to 36% of inhibition at maximum, when ether and butanol extracts were administered to rats intraperitoneally.

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Cloning and Prokaryotic Expression of C-type Lysozyme Gene from Agrius convolvuli

  • Kim, Jong-Wan;Yoe, Sung-Moon
    • Animal cells and systems
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    • v.12 no.3
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    • pp.149-155
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    • 2008
  • We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

Effect of Pretreatment with Nicotinamide on Changes in the Hepatic Metabolizing Enzyme Systme Induced by Streptozotocin (Streptozotocin에 의해 유도된 간 대사효소계의 변화에 미치는 Nicotinamide의 영향)

  • 최종원;손기호;김석환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.203-208
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    • 1991
  • The present study was undertaken in order to elucidate the effects of pretreatment with nicotinamide on changes in the hepatic metabolizing enzyme system inducted by streptozotocin (STZ). In rats, STZ(50mg/kg) administered by tail vein caused a significant rise in hepatic aniline hydroxylase and a decrease in aminopyrine N-demethylase when compared to control (p<0.05). Pretreatment with nicotinamice inhibited these effects (p<0.05). Similarly, STZ induced changes in hepatic microsomal cytochrome P-450 activity were inhibited by pretreatment with nicotinamide (p<0.05). However, changes in UDP-glucuronyl transferase and sulfortransferase activity were not significantly different(p>0.05). Pretreatment with nicotinamide also prevented STZ induced increases in glutathion S-tranferase activity when compared to the control(p<0.05). There results suggest that nicotinamide pretreatment suppresses STZ-induced changes in the hepatic metabolizing enzyme system.

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The effect of caffeine on initiation step of diethylnitrosamine-initiated hepatic altered foci in a mid-term induction system (Caffeine이 diethylnitrosamine에 의해 유도되는 preneoplastic hepatic altered foci 형성의 initiation 단계에 미치는 효과)

  • Kim, Sung-ho;Lee, Cha-soo
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.337-342
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    • 1991
  • Caffeine이 랫트의 간조직에서 diethylnitrosamine(200mg/kg B.W., DEN)에 의해 유도되는 preneoplastic altered foci형성의 initiation단계에 미치는 효과를 관찰한 바 다음과 같은 결과를 얻었다. Altered foci의 지표로 사용되는 glutathione S-transferase(GST-P)-positive foci의 수는 caffeine 음수수 ml 당 2mg병행투여군 $(7.48{\pm}3.33)$ 및 1mg 병행 투여군 $(7.50{\pm}3.32)$ 모두에서 DEN 단독투여대조군$(14.08{\pm}5.16)$에 비하여 현저히 낮게 나타났으며, 면적 또한 caffeine 2mg병행투여군 $(0.29{\pm}0.17)$, 1mg병행투여군 $(0.30{\pm}0.13)$에서 DEN단독투여대조군 $(0.46{\pm}0.21)$에 비하여 유의성 있는 낮은 수치가 관찰되었다. 이러한 결과는 caffeine이 간암발생의 initiation 단계에 작용하여 억제효과를 나타냄을 암시하였다.

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Upregulation of Glutathion S-Transferase mu 1 in Bovine Cystic Follicles

  • Kang, Da-Won;Kim, Chang-Woon;Han, Jae-Hee
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.273-279
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    • 2010
  • Follicular cystic follicles (FCFs) show delayed regression with persistent follicle growth. However, the mechanism by which follicles are persistently grown remains unclear. Glutathione S-transferases (GSTs) are drug-metabolizing and detoxification enzymes that are involved in the intracellular transport and metabolism of steroid hormones. In this study, a proteomic analysis was performed to identify whether GST expression is changed in bovine FCFs and to predict the interactions between GST and other proteins. Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and 25 mm). In bovine follicles, GST mu 1 (GSTM1) was detected as a differentially expressed protein (DEP) and significantly up-regulated in FCFs compared to normal follicles (p<0.05). Consistent with the proteomic results, semi-quantitative PCR data and western blot analysis revealed an up-regulation of GSTM1 in FCFs. Expression levels of aromatase and dehydrogenase proteins were changed in FCFs. These results show that the up-regulation of GSTM1 that is observed in bovine FCFs is likely to be responsible for the persistent follicle growth in FCFs as the activity of aromatase and the dehydrogenases.