• Title/Summary/Keyword: GST(glutathione S-transferase)

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Molecular Characterization and Expression Analysis of a Glutathione S-Transferase cDNA from Abalone (Haliotis discus hannai) (북방전복 (Haliotis discus hannai)에서 분리한 Glutathione S-transferase 유전자의 분자생물학적 고찰 및 발현분석)

  • Moon, Ji Young;Park, Eun Hee;Kong, Hee Jeong;Kim, Dong-Gyun;Kim, Young-Ok;Kim, Woo-Jin;An, Cheul Min;Nam, Bo-Hye
    • The Korean Journal of Malacology
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    • v.30 no.4
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    • pp.399-408
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    • 2014
  • Glutathione S-transferases (GSTs) are a superfamily of detoxification enzymes that primarily catalyze the nucleophilic addition of reduced glutathione to both endogenous and exogenous electrophiles. In this study, we isolated and characterized a full-length of alpha class GST cDNA from the abalone (Haliotis discus hannai). The abalone GST cDNA encodes a 223-amino acid polypeptide with a calculated molecular mass of 25.8 kDa and isoelectric point of 5.69. Multiple alignments and phylogenetic analysis with the deduced abalone GST protein revealed that it belongs to the alpha class GSTs and showed strong homology with disk abalone (Haliotis discus discus) putative alpha class GST. Abalone GST mRNA was ubiquitously detected in all tested tissues. GST mRNA expression was comparatively high in the mantle, gill, liver, and digestive duct, however, lowest in the hemocytes. Expression level of abalone GST mRNA in the mantle, gill, liver, and digestive duct was 182.7-fold, 114.8-fold, 4675.8-fold, 406.1-fold higher than in the hemocytes, respectively. Expression level of abalone GST mRNA in the liver was peaked at 6 h post-infection with Vibrio parahemolyticus and decreased at 12 h post-infection. While the expression level of abalone GST mRNA in the hemocytes was drastically increased at 3 h post-infection with Vibrio parahemolyticus. These results suggest that abalone GST is conserved through evolution and may play roles similar to its mammalian counterparts.

Effect of Cadmium on the Expression of ABC Transporters and Glutathione S-transferase in the Marine Ciliate Euplotes crassus (카드뮴이 해양 섬모충(Euplotes crassus)의 ABC Transporters와 GST 유전자 발현에 미치는 영향에 관한 연구)

  • Kim, Hokyun;Kim, Se-Hun;Kim, Ji-Soo;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.1 no.2
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    • pp.79-87
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    • 2016
  • Heavy metals such as cadmium (Cd) are highly toxic to aquatic organisms and human, even at trace concentration. Herein we investigated the effect of Cd on the gene expression of ATP-binding cassette (ABC) transporters and glutathione S-transferase (GST) in marine ciliate Euplotes crassus. Seven ABC transporters and one GST genes were partially cloned and sequences, and thereafter, transcriptional modulation of these genes after exposure to Cd for 8 h was investigated using quantitative real time RT- PCR (qRT-PCR). As results, sequence analysis and phylogenetic study revealed that E. crassus ABCs are likely typical ABC transports, in particular, B/C family, and GST gene may be similar to GST theta isoform. A significant increase in the expression of ABCs, except for ABCB21 was observed in a concentration dependent manner after exposure to Cd (0.1 and 0.5 mg/l) for 8 h. The GST mRNA level was the highest at 0.5 mg/l Cd and then reduced until control level. These findings suggest that ABCs and GST may be involved in a protective mechanism against Cd-mediated toxicity in E. crassus.

Induction of Glutathione S-transferase and NAD(P)H:Quinone Reductase by Astragali Radix Aqua-acupuncture Solution (황기(黃耆) 약침액(藥鍼液)의 Glutathione S-transferase 와 NAD(P)H: Quinone Reductase 유도)

  • Ryu Jun-Seon;Lim Jong-Kook
    • Korean Journal of Acupuncture
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    • v.18 no.1
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    • pp.21-26
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    • 2001
  • Induction of phase II enzymes such as quinone reductase (QR) or glutathione S-transferase (GST) is considered a major mechanism of protection against initiation of carcingenesis. This study was desinged to investigate the potential of Astragali Radix Aqua-acupuncture Solution (ARAS) to induce phase II enzymes and glutathione (GSH) in murine hepatoma cells grown in microtiter plate wells. ARAS was potent inducers of QR activity. ARAS was induced about 2.6-fold at concentration of $5{\times}$. In addition, GST activity was increased with ARAS. GSH levels were increased about 1.2-fold with ARAS at concentration of $0.1{\times}$. These results suggested that ARAS may act as blocking agents against carcinogenesis by induction of phase II marker enzymes.

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Increase of Salt and Low Temperature Tolerance by Overexpressing Glutathione S-Transferase (GST) Gene (염분과 저온에 대한 내성증진을 위한 GST 유전자의 과발현)

  • Jun Chol Kim;Il Seop Kim;Won Hee Kang
    • Journal of Bio-Environment Control
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    • v.11 no.3
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    • pp.139-143
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    • 2002
  • Cotton Glutathione S-Transferase (GST: EC 2.5.1.18) was cloned and overexpressed in tobacco (Nicotiana tabacum) plants. Northern blot analysis confirmed the successful transformation of cotton gst gene in tobacco plant. Type I and Type ll transcript patterns were identified in transgenic tobacco plants and only Type I transcripts were discussed in this paper, The activity of GST in the type II transgenic plants was about 1.5-fold higher than those of the wild type and non-expresser by using 1-chloro-2,4-dinitrobenzene (CDNB) and reduced glutathione as the substrate. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type II transgenic plants produced functional GST in the cells. The effects of cotton GST in the seedlings was evaluated by growing the control and transgenic seedlings at $15^{\circ}C$ in the growth chamber in the light. Overexpressors were grown well compared to the control plants (non-expressors). lo test far tolerance to salinity, seeds of Gh-5 overexpressors and the wild type Xanthi seedlings were grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings on 50 and 100 mM NaCl solution. There was no difference in growth rate at 150 and 200mM NaCl concentration.

Changes of Glutathione S-transferase Activity in MDCK Cells with Influenza Virus Type A (Influenza Virus Type A 감염 MDCK세포에 있어서 Glutathione S-transferase의 활성 변동)

  • 김병렬;박남표;윤종국;전태원;이상일
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.19-28
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    • 2000
  • This study was performed to evaluate the activities of glutathione S-transferase (GST) and lactate dehydrogenase (LDH) in Maddin-Darby canine kidney (MDCK) cells infected with virus and/or treated with amantadine. On cell morphological findings, monolayer fractions in MDCK cells infected with virus were exfolated more than 80% in 1 TCID$_{50}$ group and that in 10 TCID$_{50}$ were completely exfolated after 3 days during infectious process. In proportion to the dose of amantadine, activities of GST and LDH of MDCK cells were significantly decreased and those of LDH in medium fraction were more significantly increased compared with control. According to in both dose and time of virus innoculation, activities of GST and LDH in MDCK cells were significantly decreased in 1 and 10 TCID$_{50}$ infected cells after 3 days. LDH activities in infectious medium were remarkably rised at 10 fold. In case of the cell line inoculated with type A 100 TCID$_{50}$ and additionally treated with amantadine, the decreasing rate to the control in activities of GST and LDH was lower than that in those in case of that infected with virus only. These results suggested that virus infection and amantadine treatment may effect the activity of the detoxicating enzyme in the target cells.

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Different Sources of $\omega3$ Fatty Acids at the Fixed Ratio of p/s Affect Glutathione Dependent Enzymes in Rat Hepatocarcinogenesis (간세포 암화과정에서 p/s 고정비율과 $\omega3$ 지방산 급원에 따른 전암성 병변과 Glutathione 의존 해독화 효소계에 미치는 영향)

  • 이해정;김혜경;최혜미
    • Journal of Nutrition and Health
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    • v.36 no.8
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    • pp.785-792
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    • 2003
  • This study is conducted to determine the effects of dietary source of $\omega$3 fatty acids on preneoplastic foci and the glutathione dependent enzymes in rat hepatocarcinogenesis initiated by diethylnitrosamine (DEN). Male Sprague-Dawley rats were fed one of three diets containing 10% (w/w) fats fixed p/s = -1.0 and $\omega$6/$\omega$3 ratio = -0.4 or 4.0 ; fish oil-com oil blended (FC), com oil-beef tallow-fish oil blended (CF), com oil-beef tallow-perilla oil blended (CP), from gestation period. At 10 weeks, animals of experimental groups were injected intraperitoneally with DEN (200 mg/kg body weight) and two-thirds partial hepatectomy was carried out 3 weeks later and were sacrificed 8 weeks after DEN initiation. The area and number of glutathione S-transferase placenta (GST-P) positive foci were significantly decreased in rats fed diets containing fish oil (FC and CF) than those fed perilla oil diet (CP). Fish oil feeding significantly increased the activities of glutathione dependent enzymes. Rats fed diets containing fish oil (FC and CF) significantly increased the glutathione (GSH) content and the activities of glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST). Glutathione dependent enzymes had significantly negative correlation with GST-P positive foci. Glucose 6-phosphatase (G6Pase) was increased in rats feeding fish oil. Thiobarbituric acid reactive substances were not different among groups. Therefore, the preventive effect against hepatocarcinogenesis might be explained by induction of the glutathione dependent enzymes and G6Pase. (Korean J Nutrition 36(8): 785∼792, 2003)

Association between the Polymorphism of Glutathione S-transferase Genes and Chronic Myeloid Leukemia in Asian Population: a Meta-analysis (아시아인종에서 만성골수성백혈병과 Glutathione S-transferase 유전자 다형성의 메타분석)

  • Kim, Hee Sung
    • The Journal of the Korea Contents Association
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    • v.17 no.10
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    • pp.289-299
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    • 2017
  • To verify the association between susceptibility to chronic myeloid leukemia (CML) and GSTM1, GSTT1 gene polymorphisms in Asian populations, 9 papers published until July 2017 were cited in a meta-analysis. The null present types of the GSTM1, GSTT1 gene were analyzed individually. The significant association was found between CML and GST polymorphism (GSTM1; OR=1.306, 95% CI=1.091-1.563, p=0.004, GSTT1; OR=1.987, 95% CI=1.438-2.746, p=0.000). In addition, there was association between CML and the null type of the combination GSTM1-GSTT1 polymorphisms (OR=4.191, 95% CI=2.833-6.201, p=0.000). Thus, genetic polymorphisms of the GSTM1, GSTT1 and combination GSTM1-GSTT1 polymorphism in Asian populations may be risk factors for CML.

Comparison of Antioxidant and Glutathione S-Transferase Activities of Extracts from Acer mono and A. okamotoanum (고로쇠와 우산고로쇠 나무의 항산화능 및 glutathione S-transferase 활성 비교)

  • Jin, Ling;Han, Jae-Gun;Ha, Ji-Hye;Jeong, Hyang-Suk;Kwon, Min-Chul;Jeong, Myoung-Hoon;Lee, Hak-Ju;Kang, Ha-Young;Choi, Don-Ha;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.6
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    • pp.427-433
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    • 2008
  • This study was performed to investigate antioxidant activities and glutathione S-transferase (GST) activity according to parts of the Acer mono and A. okamotoanum. Most extracts showed high scavenging activities on DPPH. Especially, the bark of A. okamotoanum showed higher activity as 98.4% than the control, BHA as 96.5%. A. mono and A. okamotoanum showed high ability on nitrite scavenging, but decreasing tendency according to decreasing of pH. On SOD-like test, the wood of A. okamotoanum showed highest activity as 35.4% at 1.0mg/ml concentration. Also, the extracts obtained high activity on GST test. Therefore, the water extracts from the bark of A. mono and A. okamotoanum have relatively good antioxidant activity and GST activity. Especially, the bark of A. okamotoanum showed the highest activity on all of extracts, could be the use of functional foods and biomaterials.

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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Site-directed Mutagenesis of Arginine 13 Residue in Human Glutathione S-Transferase P1-1

  • Koh, Jong-Uk;Cho, Hyun-Young;Kong, Kwang-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.28 no.5
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    • pp.772-776
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    • 2007
  • In order to study the role of residue in the active site of glutathione S-transferase (GST), Arg13 residue in human GST P1-1 was replaced with alanine, lysine and leucine by site-directed mutagenesis to obtain mutants R13A, R13K and R13L. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. Mutation of Arg13 into Ala caused a substantial reduction of the specific activity by 10-fold. Km GSH, Km DCNB and Km EPNP values of R13A were approximately 2-3 fold larger than those of the wild type. Mutation of Arg13 into Ala also significantly affected I50 values of S-methyl-GSH that compete with GSH and ethacrynic acid, an electrophilic substrate-like compound. These results appeared that the substitution of Arg13 with Ala resulted in significant structural change of the active site. Mutation of Arg13 into Leu reduced the catalytic activity by approximately 2-fold, whereas substitution by Lys scarcely affected the activity, indicating the significance of a positively charged residue at position 13. Therefore, arginine 13 participates in catalytic activity as mainly involved in the construction of the proper electrostatic field and conformation of the active site in human GST P1-1.