• Title/Summary/Keyword: glutathione S-transferase activity

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Pseudomonas sp. strain DJ77에 존재하는 Glutathione S-Transferase 아미노 말단잔기의 Site-directed Mutagenesis

  • Woo, Hee-Jong;Park, Yong-Chjun;Kim, Sungje;Chung, Yong-Je;Chung, An-Sik;Kim, Young-Chang
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.374-378
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    • 1997
  • Glutathione S-transferase (GST) was purified from Pseudomonas sp. DJ77, and its N-terminal sequence was determined to be MKLFISPGACSL. A specific tyrosyl residue in the vicinity of the N terminus is conserved in all the known cytosolic GSTs and has been shown to function as a catalytic residue in $\alpha$, $\mu$, $\pi$ class GSTs from mammals. However, Pseudomonas sp. DJ77 GST has the Phe-4 and Ile-5 instead of Tyr in N-terminus. Its replacement with tyrosine did not significantly affect the enzyme activity. Results from in vitro biochemical analyses were confirmed by the in vivo activity-based CDNB growth inhibition analyses. Our results clearly indicate that GST of Pseudomonas sp. DJ77 has a novel reaction mechanism different from that of mammalian GSTs.

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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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Human Glutathione S-Transferase P1 Suppresses MEKK1-mediated Apoptosis by Regulating MEKK1 Kinase Activity in HEK293 Cells

  • Zhao, Xin;Fan, Yumei;Shen, Jiayin;Wu, Yifan;Yin, Zhimin
    • Molecules and Cells
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    • v.21 no.3
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    • pp.395-400
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    • 2006
  • Glutathione S-transferase P1 (GSTP1) plays an important role in detoxification and the metabolism of xenobiotics. Here we show that GSTP1 also regulates the MEKK1-MKK7 signaling pathway. Over-expression of GSTP1 in HEK293 cells inhibited both ${\Delta}MEKK1$- and etoposide-induced apoptosis, and inhibited procaspase-3 activation and PARP cleavage. MEKK1- induced apoptosis requires both its kinase activity and proteolytic cleavage. ${\Delta}MEKK1$ activity was inhibited by over-expression of GSTP1 in vivo and MEKK1 kinase activity was also inhibited by GSTP1 in vitro when assayed with bacterially-expressed MKK7(KM) protein as substrate. GSTP1 inhibition of etoposide-induced cell apoptosis was mainly due to its ability to suppress MEKK1 kinase activity. The glutathione-conjugating activity of GSTP1 was essential for the above effects. These findings provide insight into the mechanism by which GSTP1 protects cells from genotoxin-induced apoptosis.

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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S-(N,N-Diallyldithiocarbamoyl)-N-acetylcysteine: Synthesis and Biochemical Properties Associated with Chemoprevention (S-(N,N-Diallyldithiocarbamoyl)-N-acetylcysteine의 합성 및 발암억제와 관련된 생화학적 특성)

  • 이병훈
    • Toxicological Research
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    • v.14 no.2
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    • pp.177-181
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    • 1998
  • Dithiocarbamate and mixed disulfide containing allyl functions were designed and synthesized as putative chemopreventive agents, i.e. N,N-diallyldithiocarbamate (DATC) and S-(N,N-diallyldithiocarbamoyl)-N-acetylcysteine (AC-DATC). DATC and AC-DATC were administered and the activities of cytosolic glutathione S-transferase (GST), glutathione reductase (GR) and microsomal N-nitrosodiethylamine (NDEA) deethylase were assayed in order to test the effects of these organosulfur com-pounds on the detoxification and metabolic activation system of NDEA. The amounts of hepatic glutathione (GSH and GSSG) was also determined. The administration of DATC to rats led to an increase in the activity of GR and to an inhibition of CYP2E1-mediated NDEA deethylation. AC-DATC induced the activity of GR and GST, increased the hepatic GSH content and inhibited the rate of NDEA deethylation. The level of GSSG was decreased as a consequence of the increased activity of GR. These effects may contribute to possible antimutagenic and anticarcinogenic action of the dithiocarbamates investigated.

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Suppressive Effects of Citron Tea on Induction of Placental Glutathione S-transferase(GST-P) Positive Foci (유자차가 랫드 간암화과정에서 태반형 Glutathione S-transferase(GST-P) 양성 병소에 미치는 영향)

  • 김형숙;김희선;신길상;최혜미
    • Toxicological Research
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    • v.13 no.4
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    • pp.409-415
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    • 1997
  • The influences of dietary supplement of citron tea on the hepatocellular chemical carcinogenesis have been studied by examining placental glutathione S-transferase(GST-P) positive foci area in a liver tissue, contents of total cytochrome P450, thiobarbituric acid reactive substances(TBARS) and glucose 6-phosphatase(G6Pase) in hepatic microsome and glutathione S-transferase(GST) activity. Weaning Sprague-Dawley male rats were fed AIN76 diet with or without citron tea supplement. Rats of CTR and CTR+ groups were fed diet without citron tea supplement while CDI and CDI+ groups were fed diet with citron tea supplement for the entire experimental period(13 weeks). Rats of CDP and CDP+ groups were fed diet without citron tea supplement for the first 7 weeks and swiched to citron tea containing diet for the last 6 weeks of experimental period. CTR+, CDI+ and CDP+ groups were carcinogen treated group. Diethylnitrosamine(DEN) was used as a carcinogen initiator and injected to the rats of carcinogen treated groups as a single dose of 200 mg/kg body weight intraperitoneally after 4 weeks of feeding. 2-Acethylaminofiuorene(AAF) was used as a carcinogen promoter and supplied in the diets of carcinogen treated rats as 0.02% content for the last 6 weeks starting from 2 weeks after DEN injection. Rats were sacrificed after 13 weeks of feeding. Liver/body weight ratio and GST activities were increased by carcinogen treatment. However, they were not changed by citron tea supplement. Total cytochrome P450 contents were not changed by carcinogen treatment or citron tea supplement. TBARS contents of carcinogen treated rats showed tendency to decrease by citron tea supplement. G6Pase activity decreased by carcinogen treatment and citron tea supplement. The area of GST-P positive foci detected in carcinogen treated rats were decreased by citron tea supplement and not affected by the timing and the duration of citron tea supplement. These results suggest that citron tea has suppressive effects on hepatocellular chemical carcinogenesis probably through antioxidant compounds by decreasing TBARS contents.

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Effect of Cyclohexanone Treatment on the Serum Levels of Glutathione S-Transferase Activities in Acute Liver Damaged Rats

  • Park, Hye-Jung;Yoon, Chong-Guk
    • Biomedical Science Letters
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    • v.7 no.4
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    • pp.191-196
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    • 2001
  • To evaluate an effect of cyclohexanone (CHO) treatment on the serum levels of glutathione S-transferase (GST) activity in acute liver damaged animals, acute liver damage was induced in rats with pretreatment of 50% $CCl_4$ in olive oil (0.1 ml/100 g body wt) intraperitoneally 14 times every other day. To liver damaged rats, CHO (1.56 g/kg body wt, i. p.) was injected once and then rats were sacrificed at 4 hours after injection of CHO. Increasing rate of GST activity to the control in serum was higher in CHO-treated rats pretreated with CCL$_4$ than the $CCl_4$-pretreated those. All the more, the injection of CHO to the liver damaged rats led to more enhanced liver damage on the basis of liver functional findings, i. e., serum levels of alanine aminotransferase (ALT) activity, liver weight per body weight, and malondialdehyde content. The changing pattern of serum ALT activity was similar with that of GST activity, whereas that of liver in both enzymes differed more or less from each other; the liver GST activity in CHO-treated rats pretreated with $CCl_4$ being more increased tendency than that of $CCl_4$-pretreated rats. Concomitantly the injection of CHO showed a increasing tendency of liver GST activity compared with the control. Furthermore, CHO injection to the liver damaged rats showed somewhat higher Vmax in the kinetics of liver GST enzymes. In conclusion, injection of CHO to the liver damaged animals led to more increased activity of serum GST, and it may be chiefly caused by the alteration of membrane permeability.

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Growth-Dependent Variations in Antioxidant and Redox Enzyme Activities of Schizosaccharomyces pombe

  • Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Dae-Myung;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.3
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    • pp.278-283
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    • 2001
  • Antioxidant and redox enzyme activities are known to be involved in the cellular responses to various stresses. Their variations were observed according to the growth cycle of the fission yeast Schizosaccharomyces pombe. Peroxidase activity appeared to be notably higher in the early exponential phase than in the mid-exponential and stationary phases. However, catalase activity showed a variation pattern resembling the growth curve. Glutathione S-transferase activity was higher in the early exponential and late stationary phases. Activities of the two redox enzymes, thioredoxin and thioltransferase (glutaredoxin), were high in the stationary phase. However, their activities appeared to increase from the early exponential to mid-exponential phase. Total glutathione content had a varying pattern similar to that of thioredoxin and thioltransferase. However, its content in the early exponential phase was high. These results propose that antioxidant and redox enzymes tested are also involved in the mechanism of cell growth.

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Effect of Taurine Supplementation on Hepatic Lipid Peroxide Metabolism in Streptozotocin-induced Diabetic Rats

  • You, Jeong-Soon;Chang, Kyung-Ja
    • Journal of Community Nutrition
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    • v.2 no.2
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    • pp.164-169
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    • 2000
  • The purpose of this study was to examine the effect of taurine supplementation time on the activity of the enzymes metabolizing lipid peroxide in the liver of streptozotocin(STZ)-induced diabetic rats, Sprague-Dawley male rats were fed the purified diet for 7 weeks. They were supplemented with or without 1% taurine in drinking water before or after STZ injection or during all experimental procedure. In comparison to diabetic group without taurine, glucose-6-phosphatase(G6Pase) activity was decreased in diabetic group supplemented with taurine before STZ injection, and it was increased in diabetic group supplemented with taurine after STZ injection, but the difference was not significant. Glutathione S-transferase(GST) activity was significantly increased by the injection of STZ. However, the GST activities of diabetic groups exposed to taurine after STZ injection or during all experimental procedure were significantly decreased. Glutathione peroxidase(GPx) activities was significantly decreased by STZ injection. However, only in diabetic group supplemented with taurine before STZ injection, GPx activities was not decreased by the STZ injection. These results suggest that taurine supplementation may change the activities of GSH-related enzymes metabolizing lipid peroxide in the liver of streptozotocin(STZ)-induced diabetic rats and that may be helpful for the prevention of diabetic complication.

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