• Title/Summary/Keyword: Glutathione S-Transferase

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Generation and characterization of a monoclonal antibody with high species-specificity to Schistosoma japonicum glutathione S-transferase

  • Kim, Jung-Hwan;Park, Jung-Hyun;Ju, Sung-Kyu;Lee, Myung-Kyu;Kim, Kil Lyong
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.187-195
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    • 2001
  • The expression of recombinant proteins fused to 26 kDa glutathione S-transferase (GST) extracted from Schistosoma japonicum represents an attractive system for purifiying proteins of interest in a single step using GST-affinity chromatography. In addition, the GST-tag is used conveniently for detecting fused proteins since its high solubility as well as its relatively small size rarely interferes with the biological activity of the fused protein. In this regard, the GST system is frequently applied for tracing fusion proteins in both prokaryotic and eukaryotic cells to elucidate the physiological interactions and functional compartments of proteins. To provide a further tool in analyzing GST-fusion proteins, a new monoclonal antibody, with a high specificity to the S. japonicum GST was produced. Methods: BALB/c mice were immunized both with recombinant S. japonicum GST proteins, and by the fusion of splenocytes from these mice with myeloma cells. From this, a new anti -GST monoclonal antibody, termed SARAH, was generated. The specificity and reactivity of this antibody was confirmed by ELISA and by Western blot analysis. Results: SARAH showed a high reactivity to recombinant GST and GST fusion protein but not with native mammalian GST proteins as derived from other species including humans, cows, rabbits and rats. The applicability of SARAH was further demonstrated by confocal laser scanning microscopy, where GST proteins that were expressed transiently in mouse fibroblast cells, were specifically detected without interference of endogenous GST. Conclusion: SARAH is new monoclonal antibody with a high specificity to recombinant GST proteins but not to endogenous GST in mammalian cells.

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Preliminary X-ray Studies of a New Crystal form of 28 kDa Clonorchis sinensis Glutathione S-Transferase

  • Cho, Youn-Hye;Kim, Young-Kwan;Kim, Seung-Joon;Hong, Seong-Jong;Chung, Yong-Je
    • Korean Journal of Crystallography
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    • v.16 no.2
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    • pp.138-140
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    • 2005
  • A new crystal of helminth glutathione S-transferase, 28 kDa isozyme from Clonorchis sinensis has been grown from a 20% PEG MME 550 solution containing 50 mM $CaCl_{2}$ in 0.1 M bis-Tris buffer (pH 6.5) in $2{\sim}3$ days. The crystals diffract to $3.0{\AA}$ resolution and belong to the orthorhombic space group $P2_{1}2_{1}2_{1}$ with cell parameters $a=62.58{\AA},\;b=69.92{\AA},\;and\;c=339.67{\AA}$.

Ingredients of Tyrosinase and Elastase Inhibitory Activity from Calyx of Diospyros kaki Thunberg (감꼭지의 Tyrosinase와 Elastase 저해 활성 성분)

  • Cha, Bae Cheon;Lee, Eun Hee
    • Korean Journal of Pharmacognosy
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    • v.44 no.2
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    • pp.182-187
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    • 2013
  • This study was conducted to investigate glutathione S-transferase(GST) activity, tyrosinase inhibitory effect and elastase inhibitory effect in persimmon calyx(calyx of Diospyros kaki Thunberg) for screening of functional materials from natural products. As a result, EtOAc extract of persimmon calyx turned out to be having tyrosinase inhibitory effect and elastase inhibitory effect. The active constituents of tyrosinase and elastase inhibitory effect were isolated from EtOAc extract of persimmon calyx. Their structure of compounds were identified as ursolic acid and (-)-daucosterol by spectroscopic evidence, respectively.

Effects of Bombyx mori Larvae Extracts on Carbon Tetrachloride-Induced Hepatotoxicity in Mice (사염화탄소에 의해 유발된 생쥐의 간독성에 미치는 누에 추출물의 영향)

  • 류강선;이희삼;김선여
    • Journal of Life Science
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    • v.9 no.4
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    • pp.375-381
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    • 1999
  • To evaluate the protective effects of extracts of silkworm on carbon tetrachloride-induced hepatotoxicity, serum glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase and lactic dehydrogenase activities, malondialdehydes values and glutathione-S-transferase activity were measured in ICR mice. Extracts of silkworm was administered orally at 30min after the administration of CCl4 Mice were sacrificed at 24h after the administration of extracts of silkworm. The activities of serum aminotransferase and the hepatic content of lipid peroxide after carbon tetrachloride treatment were markedly increased than normal control but those levels were decreased by the treatment of butanol soluble fraction of silkworm methanol extract. Glutathione S-transferase activity was decreased by carbontetrachloride than control, but also inhibited by the treatment of butanol soluble fraction of silkworm methanol extract.

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The effect of Korean ginseng on diethylnitrosamine-initiated hepatic altered foci in a mid-term induction system (고려인삼이 diethylnitrosamine에 의한 유도되는 preneoplastic hepatic altered foci의 형성에 미치는 효과)

  • Kim, Sung-ho
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.45-50
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    • 1989
  • 홍삼 및 수삼이 랫트의 간조직에서 diethylnitrosamine(DEN)에 의해 유도되는 preneoplastic altered foci 형성에 미치는 영향을 관찰한 바 다음과 같은 결과를 얻었다. Altered foci의 지표로 사용되는 glutathione S-transferase (GST-P)-positive foci의 숫자는 DEN 단독투여군($9.07{\pm}5.69$)에 비하여 수삼병행 투여군 ($4.77{\pm}3.23$)에서, 면적은 DEN 단독투여군($0.93{\pm}0.65$)에 비하여 홍삼병행투여군 ($0.50{\pm}0.31$)에서 각각 현저한 감소를 나타냈다. 이러한 결과는 홍삼 및 수삼이 간암 발생을 억제하는 작용이 있음을 암시하였다.

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Protective Effects of Extracts of Mori Cortex Radicis on Carbon Tetrachloride-induced Hepatotoxicity (사염화탄소에 의해 유발된 간독성에 대한 상백피 추출물의 간보호효과)

  • 김선여;이희삼;류강선;이은주;김영중
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.391-396
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    • 1999
  • In order to evaluate the protective effects of extracts of Mori cortex radicis on carbon tetrachloride-induced hepatotoxicity, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactic dehydrogenase, malondiadehyde values and glutathione S-transferase activity were measured in ICR mice. The activities of serum aminotransferase and the hepatic content of lipid peroxide after carbon tetrachloride-treatment were markedly increased than normal control but those levels were significantly decreased by the treatment of butanol fraction of Mori cortex radicis mathanol extract. Glutathione S-transferase activity was decreased by carbon tetrachloride than control, but that also inhibited by the treatment of butanol fraction of Mori cortex radicis methanol extract. These results demonstrate a possible hepatoprotective role of extract role of extract of Mori cortex radicis against ${CCl_4}-induced$ hepatoxicity in vivo.

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PRELIMINARY X-Ray DIFFRACTION STUDY OF Pseudomonas sp. DJ77 GLUTATHIONE S-TRANSFERASE

  • Park, Heung-Soo;Chung, An-Sik;Ryu, Seong-Eon;Suh, Se-Won;Kim, Young-Chang;Chung, Yong-Je
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.20-20
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    • 1996
  • Crystals of a bacterial glutathione S-transferase(pGST) from pseudomonas sp. DJ 77 have been grown by hanging drop method of vapour diffusion from ammonium sulfate solution. The low concentration of polyethylene glycol 400 as additive were found to be essential for the reproducible growth of large single crystal of pGST. (omitted)

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Crystallization and Preliminary Crystallographic Study of 26kDa Clonorchis sinensis glutathione S-transferase Complexed with Inhibitors

  • Chung, Yong-Hak;Chung, Yong-Je
    • Korean Journal of Crystallography
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    • v.11 no.4
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    • pp.238-240
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    • 2000
  • A helminth glutathione S-transfease, 26 kDa isozyme from Clonorchis sinensis was cocrystallized with inhibitors by the hanging-drop method with monomethylether 550 as a precipitant. The crystals, grown in the presence of S-methylglutathione or trans-4-phenyl-3-buten-2-one, suitable for X-ray analysis, belong to the hexagonal space group P6₂(or P6₄), with unit cell parameters a= b= 97.6Å, c=117.4Å. X-ray diffraction data were collected with 2.5Å resolution.

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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.