• Title/Summary/Keyword: GST

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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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Target extraction using divergent-direction-emphasis symmetry transform (발산 방향성 강조 대칭변환을 이용한 표적 검출)

  • Jun, Jun-Hyung;Lee, Hee-Yul;Choi, Byung-Jae;Park, Kil-Houm
    • Journal of the Korean Institute of Intelligent Systems
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    • v.20 no.5
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    • pp.665-671
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    • 2010
  • This paper propose the DDEGST(divergent-direction-emphasis generalized symmetry transform) which emphasis the symmetry of divergent intensity orientation for effective target extraction in FLIR(forward looking infra-red) images. In the proposed method, we use the exponential function instead of cosine function as a phase weight function in the generalized symmetry transform for effective target extraction in FLIR images which contain a target with higher intensity than a background intensity. To evaluate the performance of the proposed method, we compare the proposed mehtod with conventional GST method in experiments. We prove that the proposed method have better performance in IR images.

In vitro Interaction of Recombinantly Expressed Kringle 5 (rK5) with Ras Guanine Nucleotide Dissociation Stimulator-like Factor (Rgl2)

  • Lee, Jung-Whoi;Kim, Sun-Hee;Park, Yong-Sung;Woo, Je-Wan;Lim, Dong-Yeol;Lee, Kyung-Hee
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1863-1868
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    • 2004
  • Kringle 5 (K5), located outside of angiostain (K1-4) in human plasminogen, displays more potent antiangiogenic activity on endothelial cell proliferation than angiostatin itself. Using a yeast two-hybrid system in vivo, we have recently identified Rgl2 (guanine nucleotide dissociation stimulator (RalGDS)-like factor 2) as a binding protein of human K5. In order to confirm in vitro protein interaction between K5 and Rgl2, we developed bacterial recombinant expression systems for them. K5 and Rgl2 proteins were expressed in high yields and purified into pure forms with His tags and GST fusion, respectively. GST-pull down experiments clearly demonstrated that K5 interacts specifically with Rgl2 in vitro. These results indicate that Rgl2 functions as a receptor protein for K5 in vitro as well as in vivo, leading to anti-angiogenesis through regulating Ras signaling pathways.

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.

Properties of GST Thin Films for PRAM with Composition (PRAM용 GST계 박막의 조성에 따른 특성)

  • Jung, Myung-Hun;Jang, Nak-Won;Kim, Hong-Seung;Ryu, Sang-Ouk;Lee, Nam-Teal;Yoon, Sung-Min;Park, Young-Sam;Lee, Seung-Yun;Yu, Byoung-Gon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2005.07a
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    • pp.203-204
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    • 2005
  • PRAM (Phase change Random Access Memory) is one of the most promising candidates for next generation Non-volatile Memories. The Phase change material has been researched in the field of optical data storage media. Among the phase change materials $Ge_2Sb_2Te_5$(GST) is very well known for its high optical contrast in the state of amorphous and crystalline. However, the characteristics required in solid state memory are quite different from optical ones. In this study, the structural properties of GST thin films with composition were investigated for PRAM. The 100-nm thick GeTe and $Sb_2Te_3$ films were deposited on $SiO_2$/Si substrates by RF sputtering system. In order to characterize the crystal structure and morphology of these films, we performed x-ray diffraction (XRD) and atomic force microscopy (AFM).

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Induction of Hepatic Glutathione S-transferase Activity in Mice Administered with Various Vegetable Extracts

  • Kim, Mee-Ree;Lee, Kun-Jong;Kim, Yun-Bae;Sok, Dai-Eun
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.207-213
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    • 1997
  • The effect of various vegetables commonly consumed by Koreans on the induction of glutathione S-trasferase(GST) activity in mice was assessd. The extract of vegetable dissolved in propylene glycol (5ml/kg body wt.) was administered to ICR female mice 6 to 8 weeks old via gavage during 5 days. The changes of body weight and liver weight of all treated groups were not significantly different compared with control group. Hepatic protein contents of trated groups compared with control group were not significantly different except BHT treated group. The induction of GST activity in liver cytosol of mice was the greatest with broccoli, followed by radish, wild green onion, turnip, and green onion. The induction of GST activity in liver cytosol increased up to 1.5 to 1.8-folds at a dose of 24 g fresh vegetable/mouse. The induction of combination between vegetables was the highest with the combination of broccoli and radish (1.83-fold), followed by that of broccoli and green onion (1.72-fold), and that of broccoli and turnip (1.50-fold).

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Functional Studies of Cysteine Residues in Human Glutathione S-Transferase P1-1 by Site-Directed Mutagenesis

  • Park, Hui Jung;Lee, Gwang Su;Gong, Gwang Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.1
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    • pp.77-83
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    • 2001
  • To gain further insight into the relationship between structure and function of glutathione S-transferase (GST), the four cysteine mutants, C14S, C47S, C101S and C169S, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized glutathione (GSH). The catalytic activities of the four mutant enzymes were characterized with five different substrates as well as by their binding to four different inhibitors. Cys14 seems to participate in the catalytic reaction of GST by stabilizing the conformation of the active-site loop, not in the GSH binding directly. The substitution of Cys47 with serine significantly reduces the affinity of GSH binding, although it does not prevent GSH binding. On the other hand, the substitution of Cys101 with serine appears to change the binding affinity of electrophilic substrate by inducing a conformational change of the $\alpha-helix$ D. Cys169 seems to be important for maintaining the stable conformation of the enzyme. In addition, all four cysteine residues are not needed for the steroid isomerase activity of human glutathione S-transferase P1-1.

Effect of Dietary Zinc Levels on Cadmium-Induced Hepatotoxicity in Rats (식이성 아연 수준이 카드뮴 중독에 미치는 영향)

  • 조수열;김명주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.4
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    • pp.574-580
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    • 1994
  • The effect of dietary zinc(Zn) levels on cadmium (Cd)-induced hepatotoxicity was studied in serum and liver of rats. Adult male Spraque-Dawley rats were fed on diets containing one of three levels of zinc carbonate(0, 56, $560\mu\textrm{g}/kg$ diet) and Cd-treated groups were administrated oral intubation with cadmium chloride 95.0 mg/kg of body weight) at the sametime once a week. Net weight gain (NWG), feed intake (FI) and feed effciiency ratio (FER) in Zn deficiency groups significantly decreased as compared to that of control and excessive groups. Cd oral intubation caused a decrease in NWG and FI but an increase in Zn deficiency group in FER. GSH-Px, GST and catalase activity showed significant decrease in Zn deficiency and Zn excessive group. LPO content in liver significantly increased in Zn deficiency group. Cd oral intubation increased the content of LPO in Zn deficiency group as compared to control. GSH content and GST activity of hepatic tissue significantly decreased in Zn deficiency and excessive group. The activity of AST and ALT in serum were markedly increased in Zn deficiency, Zn excessive and Cd-treated groups. LDH and ALP activities significantly increased in Cd-treated group while ALP activity decreased by Zn deficiency. It was observed that the livers of rats exposed to Cd and Zn excessive group showed a marked increase of hepatic enzyme as compare to only Cd-treated in rats.

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Characterization of the Monoclonal Antibody Specific to Human S100A2 Protein (인체 S100A2 단백질에 특이적인 단일클론 항체)

  • Kim, Jae Wha;Yoon, Sun Young;Kim, Joo Heon;Joo, Jong-Hyuck;Kim, Jin Sook;Lee, Younghee;Yeom, Young Il;Choe, Yong-Kyung;Choe, In Seong
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.16-22
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    • 2003
  • Background: The S100A2 gene, also known as S100L or CaN19, encodes a protein comprised of 99-amino acids, is a member of the calcium-binding proteins of EF-hand family. According to a recent study, this gene was over-expressed in several early and malignant carcinomas compared to normal tissues. To elucidate the role of S100A2 protein in the process during carcinogenesis, production of monoclonal antibody specific to the protein is essential. Methods: First, cDNA sequence coding for ORF region of human S100A2 gene was amplified and cloned into an expression vector to produce GST fusion protein. Recombinant S100A2 protein and subsequently, monoclonal antibody to the protein were produced. The specificity of anti-S100A2 monoclonal antibody was confirmed by immunoblot analysis of cross reactivity to other recombinant proteins of S100A family (GST-S100A1, GST-S100A4 and GST-S100A6). To confirm the relation of S100A2 to cervical carcinogenesis, S100A2 protein in early cervical carcinoma tissue was immunostained using the monoclonal antibody. Results: GST-S100A2 recombinant protein was purified by affinity chromatography and then fusion protein was cleaved and S100A2 protein was isolated. The monoclonal antibody (KK0723; Korean patent pending #2001-30294) to the protein was produced and the antibody did not react with other members of EF-hand family proteins such as S100A1, S100A4 and S100A6. Conclusion: These data suggest that anti-S100A2 monoclonal antibody produced in this study can be very useful for the early detection of cervical carcinoma and elucidation of mechanism during the early cervical carcinogenesis.

Chemopreventive Effects of the Extracts from Soybean Fermented with Basidiomycetes (들송이(Agrocybe cylindracea)와 말똥진흙버섯(Phellinus igniarius)을 배양한 대두추출물의 암예방 효과)

  • Kim, So-Yeun;Son, Jun-Ho;Ha, Hyo-Cheol;Lee, Hang-Woo;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.124-130
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    • 2002
  • The inductions of phase II enzymes, such as NAD(P)H : quinone reductase (QR), glutathione S-transferase (GST), glutathione (GSH) level and the inhibition of polyamine metabolism were tested for the chemopreventive potentials of the extracts from the soybean fermented with Agrocybe cylindracea (AC) or Phellinus igniarius (PI). The soybean fermented with AC or PI was potent inducer of QR activity in murine hepatoma Hepa1c1c7 cells. GST activities of the extracts from soybean fermented with AC or PI were higher than that of the extract from soybean not fermented with basidiomycetes. In addition, GSH levels of the extracts from soybean fermented with AC or PI were increased about 1.2 fold or 1.4 fold, respectively. In addition, proliferation of Acanthamoeba castellanii in a broth medium was inhibited by the extracts from soybean fermented with AC or PI at the both concentration of 20 and 40 mg/3 ml. These results suggest that soybean fermented with AC or PI may have chemopreventive potentials by inducing QR activity, increasing GSH and GST levels and inhibiting polyamine metabolism.