• Title, Summary, Keyword: glutathione S-transferase

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Inhibition of glutathion-S-transferase and amidase by impurities in technical grade benfuracarb (Benfuracarb 원제에 함유된 불순물들의 glutathione-S-transferase와 amidase 저해 특성)

  • Yum, Chang-Sub;Kim, Song-Mun;Yu, Ji-Sook;Hur, Jang-Hyun
    • The Korean Journal of Pesticide Science
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    • v.6 no.1
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    • pp.31-35
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    • 2002
  • Objectives of this study were to determine if impurities in technical grade benfuracarb inhibit glutathione-S-transferase and amidase and to identify structures of impurities in technical grade benfuracarb. Technical grade benfuracarb, active ingredient, and impurity inhibited glutathione-S-transferase, and their $I_{50}$ were $9.7{\times}10^{-4}M,\;>1.0{\times}10^{-3}M,\;1.8{\times}10^{-4}M$, respectively. Such inhibition, however, was not higher than that by ethacrynic acid, a selective inhibitor to GST. Technical grade benfuracarb, active ingredient, and impurity also inhibited amidase, and their $I_{50}$ were $6.0{\times}10^{-5}M,\;4.3{\times}10^{-4}M,\;7.6{\times}10^{-5}M$, respectively. Our results show that the inhibition of both detoxifying enzymes by impurities in benfuracarb was 10-fold lower than that by active ingredient, suggesting that both active ingredient and impurities are involved in the inhibition of both detoxifying enzymes. Of four impurities (IM $1{\sim}4$) that were separated from technical grade benfuracarb, IM 2 and IM 3 inhibited GST and amidase. Based on data from IR, $^1H$-NMR, $^{13}C$-NMR and MS, it was determined that IM 2 is ethyl-N-isopropylamino propionate and IM 3 is ethyl-N-isopropyl-N(chlorosulfenyl)aminopropionate.

Characterization of a Novel Glutathione S-Transferase from Pseudomonas sp. DJ77

  • Jung, U-Hee;Cho, Young-Sik;Seong, Hark-Mo;Kim, Seong-Jae;Kim, Young-Chang;Chung, An-Sik
    • BMB Reports
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    • v.29 no.2
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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 $45^{\circ}C$.

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Excess Taurine Induced Placental Glutathione S-transferase Positive Foci Formation in Rat

  • Kweon, Sang-Hui;Kim, Yoon;Choi, Hay-Mie;Kwon, Woo-Jung;Chang, Kyung-Ja
    • BMB Reports
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    • v.33 no.6
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    • pp.469-475
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    • 2000
  • The purpose of this study was to examine the chemopreventive potential of taurine at various levels on the diethylnitrosamine (DEN)·induced hepatocarcinogenesis. Male Sprague-Dawley rats were fed on diets containing 0, 1, 2, 3% taurine or 5% ${\beta}-alanine$ for taurine depletion. Then they were treated with DEN and 2/3 partial hepatectomy. The number of placental glutathione S-transferase positive ($GST-P^+$) foci, as a preneoplastic marker in the 1 % taurine group was lower than the control diet group. However the difference was insignificant. Although taurine diets reduced the thiobarbituric acid reactive substance (TBARS) level, the number of $GST-P^+$ foci was increased in 3% taurine diet group. The 1 % taurine diet increased the glutathione (GSH) level and GST activity, however they unfortunately did not suppress the foci formation. In the 3% taurine group, the GSH level and GSH peroxidase (GPx) activity were significantly decreased. Excess taurine supplementation of the pharmaceutical dose worked against hepatic chemoprevention, which might result from modulation of GPx activity and GSH utility. On the contrary, taurine might work as an antioxidant against TBARS production as the 1 % taurine diet increased GSH level. The potency of the cancer preventive effect of taurine still remains and further studies should investigate the effect of taurine with less than 1 % levels on the prevention of hepatic cancer.

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The Effect of Green Tea on the Lipid Composition of Serum and Liver and the Activities of Antioxidative Enzymes in Rats (녹차가 흰쥐의 혈청 및 간의 지질성분과 항산화계 효소 활성도에 미치는 영향)

  • 정희정;유영상
    • Journal of the East Asian Society of Dietary Life
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    • v.6 no.1
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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.

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Preliminary X-Ray Diffraction Study of Glutathione S-Transferase from Pseudomonas sp. DJ77

  • Choi, Heung-Soo;Woo, Ju-Rang;Lee, Jung-Hee;Chung, An-Sik;Ryu, Seong-Eon;Kim, Young-Chang;Chung, Yong-Je
    • BMB Reports
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    • v.30 no.4
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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).

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Inhibitory Substances of a Tau-Type Pumpkin Glutathione S-Transferase: Their Existence and Chemical Properties

  • Hossain, Md. Daud;Suzuki, Toshisada;Fujita, Masayuki
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.117-122
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    • 2007
  • Distributions of physiological inhibitors of a tau-type pumpkin glutathione S-transferase(CmGSTU3) have been investigated in different organs of pumpkin plants, including the onion bulb and water hyacinth root. Inhibitory effects were observed in alcoholic extracts of all plant parts, but the extracts prepared from the roots of either water hyacinth or pumpkin plant showed the highest effect on CmGSTU3 toward 1-chloro-2,4- dinitrobenzene(CDNB). Results of various chromatographies indicated that a number of inhibitory substances were present in the alcoholic extract of each plant organ. Some macromolecules in the plant extracts exhibited inhibitory effects; however, the extracts might contain a large number of unknown low-molecular-weight inhibitory substances. Some of the low-molecular-weight inhibitors in water hyacinth root extract showed characteristics fluoresce under UV light.

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Effects of Phloroglucinol Isolated from Ecklonia stolonifera on the Acetaminophen-Metabolizing Enzyme System in Rat (해조류 곰피로부터 분리한 Phloroglucinol이 흰쥐의 아세트아미노펜 대사효소활성에 미치는 영향)

  • 박종철;허종문;박주권;김현주;전순실;최재수;최종원
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.3
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    • pp.448-452
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    • 2000
  • 실험동물에서 곰피로부터 분리한 phlorglucinol은 acetaminophen의 투여로 현저히 증가된 간조직에 있어서 지질과선화의 함량을 억제하였다. Acetaminophen 투여에 따른 간 cytochrome P-450, aminopyrine N-deme-thylase 및 aniline hydroxylase 활성변동은 관찰할 수 없었다. 곰피 성분 투여군은 glutathione S-transferase의 활성에서는 대조군의 수준에는 미치지 않으나 효소의 활성이 acetaminophen 단독 투여군보다 현저히 증가되었다. 그리고 간조직중 glutathione의 함량은 phlorglucionl을 전처리군에서 acetaminophen 단독 투여군보다 증가되었다. Glutathione reductase 활성에서는 acetaminophen 투여군은 대조군보다 활성이 감소되었으며, 성분으로 전처리한 군은 acetaminophen 단독 투여군보다 증가 되었다. 따라서 곰피에서 분리한 페놀성화합물인 phloroglucinol은 acetaminophen 투여로 증가되던 지질과 산화함량을 감소시키며, acetaminophen 대사효소활성에서는 glutathione S-transferase의 활성이 증가되어 acetaminophen 의 대사를 촉진시키는 것으로 추정된다.

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Isolation of Gglutatihone S-Ttransferase(ClGST) Gene from Codonopsis lanceolata (더덕에서 Glutathione S-transferase (ClGST) 유전자의 분리)

  • Kim Jin-Ju;Yang Deok-Chun
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.240-245
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    • 2005
  • A cDNA clone homologous to glutathione S-transferase gene was isolated and characterized from Codonopsis lanceolata(ClGST). The ClGST is 761 nucleotides long and has an open reading frame of 522 bp with a deduced amino acid sequence of 173 residues. The ClGST shows meaning homology to A. thaliana(AAC63629) $71\%$, C. chinense(CAI51314) $73\%$, E. esula(AAE65767) $75\%$, H. muticus(CAA55039) $70\%$, N. plumbaginifolia(CAA96431) $77\%$, S. commersonii(AAB65163).

Active-Site Mutants of Human Glutathione S-Transferase P1-1: Effects of the Mutations on Substrate Specificity and Inhibition Characteristics

  • Park, Hee-Joong;Yoon, Suck-Young;Kong, Kwang-Hoon
    • BMB Reports
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    • v.31 no.4
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    • pp.399-404
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    • 1998
  • In order to gain further insight on the relationship between structure and function of glutathione S-transferase (GST), the six active-site mutants, R13T, K44T, Q51A, Q64A, S65A, and D98A, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The active-site mutants showed marked differences in substrate specificity. The substitution of Gln51 with threonine resulted in a drastic decrease in the specific activities to <10% of the wild-type value. The substitution of Arg13 with threonine resulted in more decreased specific activity toward cumene hydroperoxide and in the $I_{50}$ values of S-(2,4-dinitrophenyl) glutathione and benanstatin A. These results suggest that the substitution of Arg13 with threonine changes the conformation of the active site to increase the affinity for the product or electrophilic substrate. Lys44 seems to be in the vicinity of the H-site of hGST P1-1 or may contribute to some extents to the electrophile binding.

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Antioxidant Activities of Citrus unshiu Extracts obtained from Different Solvents (추출용매에 따른 진피 추출물의 항산화 활성)

  • Lee, Sung-Gu;Oh, Sung-Cheon;Jang, Jae-Seon
    • The Korean Journal of Food And Nutrition
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    • v.28 no.3
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    • pp.458-464
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    • 2015
  • In this study, the total polyphenol content, electron donating ability (EDA) and inhibitory activity of glutathione S-transferase (GST) of freeze-dried Citrus unshiu extracts were examined. The Citrus unshiu extracts was obtained from four solvents such as ethyl acetate, acetone, methyl chloride and methanol, to evaluate its functional properties. Total polyphenol contents were measured in the two different extracts, and the extracts were screened for their potential antioxidant activities using tests such as electron donating ability (EDA), glutathione S-transferase (GST). The total polyphenol contents of Citrus unshiu extracts were $928.48{\pm}1.19{\mu}g\;GAE/mL$ in ethyl acetate (EA), $886.03{\pm}0.44{\mu}g\;RE/mL$ in acetone (AC), $413.08{\pm}1.39{\mu}g\;GAE/mL$ in methylene chloride (MC), $12,648.60{\pm}0.56{\mu}g\;GAE/mL$ in methanol (MeOH), respectively. Also, the total polyphenol contents of EtOH Citrus unshiu extracts were $664.64{\pm}0.74{\mu}g\;GAE/mL$ in EA, $702.67{\pm}0.85{\mu}g\;RE/mL$ in AC, $429.64{\pm}0.61{\mu}g\;GAE/mL$ in MC, $16,108{\pm}0.73{\mu}g\;GAE/mL$ in MeOH, respectively. The total polyphenol contents were significantly difference (p<0.05) between the solvents. The electron donating ability of Citrus unshiu extracts were $62.80{\pm}0.36%$ in EA, $97.43{\pm}0.51%$ in AC, $52.20{\pm}0.30%$ in MC, $97.63{\pm}0.46%$ in MeOH, respectively. Also, the electron donating ability of EtOH Citrus unshiu extracts were $51.49{\pm}0.26%$ in EA, $63.17{\pm}0.31%$ in AC, $67.68{\pm}0.55%$ in MC, $96.18{\pm}0.41%$ in MA, respectively. The electron donating ability were significantly difference (p<0.05) between the solvents. The inhibitory activity of glutathione S-transferase in Citrus unshiu extracts were $76.22{\pm}0.65%$ in EA, $31.73{\pm}0.48%$ in MC, $97.48{\pm}0.56%$ in MeOH, respectively. Also, inhibitory activity of glutathione S-transferase in EtOH Citrus unshiu extracts were $75.54{\pm}0.55%$ in EA, $73.53{\pm}0.38%$ in MC, $48.70{\pm}0.46%$ in MeOH, respectively. The inhibitory activity of glutathione S-transferase were significantly difference (p<0.05) between the solvents. These results indicated that the Citrus unshiu extracts is a high-valued food ingredient and the extraction with methanol will be useful as a nutritional source with natural antioxidant activities. Considering high consumer demand beneficial health effects, Citrus unshiu extracts can be utilized to develop functional food health- promoting and natural antioxidant agents.