• Title/Summary/Keyword: GST

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SPR-based Antibody-Antigen Interaction for Real Time Analysis of Carbamate Pesticide Residues

  • Yang, Gil-Mo;Kang, Suk-Won
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.15-19
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    • 2008
  • This research was conducted to develop a quick and sensitive method of detecting carbamate residues using the immobilization of antibody-antigen interactions with surface plasmon resonance (SPR). We have used commercialized surface plasmon resonance equipment (Biacore 3000). The antibody used for the immunoassay was specific for glutathione-s-transferase (GST) and the antigens included several carbamate pesticides (carbofuran, carbaryl, and benfuracarb). When antigens were applied to the protein GST, the detection limit was 2 ng/mL of carbamate pesticide. The fabricated protein GST maintained its activity for over 200 measurements. Thus we determined that the SPR biosensors could detect the specific reversible binding of a reactant in solution to a binding partner immobilized on the surface of the sensor and allow real-time detection and monitoring.

Effect of Bromobenzene Pretreatment on the Hepatic Glutathione Content and Glutathione S-transferase Activity in Bromobenzene Treated Rats (흰쥐에 있어서 Bromobenzene전처치가 간조직 중 Glutathione 및 Glutathione S-transferase활성에 미치는 영향)

  • 신중규
    • Journal of Environmental Health Sciences
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    • v.23 no.2
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    • pp.83-88
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    • 1997
  • To evaluate the effect of bromobenzene pretreatment on the bromobenzene metabolism, the animal group was induced the stage of slight liver damage with 7 times bromobenzene injection every two days (400 mg/kg body wt. i.p.). In the present experimental animal model, the single dose of bromobenzene(400 mg/kg body wt. i.p.) was injected to the bromobenzene-pretreated rats and the hepatic aniline hydroxylase(AH) activity, glutathione(GSH) content and glutathione S-transferase (GST) activity were determined at the intervals of 2, 4, 8, 24 hours throughout 24 hr. The activities of hepatic AH and GST were generally higher in bromobenzene-pretreated rats than those in normal group throughout the whole course of experiment. Furthermore, the decreasing rate of hepatic GSH content was also higher in bromobenzene pretreated rats than in normal rats. Moreover, the value of V$_{max}$ in hepatic GST was higher in bromobenzene pretreated rats than that in the normal rats. In conclusion, these results indicate that the pretreatment of bromobenzene may rather enhance the bromobenzene metabolism.

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Expression and Purification of Delta Sleep-Inducing Peptide in Escherichia coli

  • Oh, Kwang-Seok;Na, Do-Kyung;Kweon, Mee-Hyang;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.620-623
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    • 2003
  • The delta sleep-inducing peptides (DSIP, Trp-Ala-Gly-Gly-Asp-Ala-Ser-Gly-Glu) is an important regulatory hormone, controlling hypothalamus and pituitary functions. In the current study, an expression system was designed for the rapid and economic expression oi recombinant DSIP for biophysical studies. Artificially synthesized oligonucleotides encoding DSIP were cloned into a pGEX-KG vector and expressed in E. coli (BL21). The recombinant GST-DSIP was then readily purified using a GST affinity column. To obtain intact DSIP from the GST-DSIP, thrombin cleavage and a CNBr reaction were successively carried out. The DSIP in the CNBr reaction mixture was subjected to RP-HPLC purification to yield 1.2 mg DSIP from a 1 liter culture of E. coli. Identification of the DSIP was peformed using MALDI-MS and an amino acid composition analysis.

Glutathione S-transferase Activity and Hyaluronidase Inhibitory Effect of Medicinal Plants (생약의 Glutathione S-transferase 활성과 Hyaluronidase 저해효과)

  • Lee, Eun-Hee;Cho, Jae-Yong;Cha, Bae-Cheon
    • Korean Journal of Pharmacognosy
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    • v.35 no.3 s.138
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    • pp.184-188
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    • 2004
  • This study was carried out to evaluate glutathione S-transferase (GST) activity and hyaluronidase inhibitory effect of medicinal plants. The EtOH extracts of 20 species plants were tested. As the result, Acorus gramineus and Pueraria lobata exhibited GST activity. On the continuous experiment, the n-BuOH fraction of Acorus gramineus and the $H_2O$ fraction of Pueraria lobata showed the elevation of GST activity. On the experiment of hyaluronidase inhibitory effect, Acorus gramineus exhibited a potent inhibitory activity. These results suggest that the extract of Acorus gramineus can be applicable for the development of a new anti-inflammatory agent.

The Phase Transition with Electric Field in Ternary Chalcogenide Thin Films

  • Yang, Sung-Jun;Lee, Jae-Min;Shin, Kyung;Chung, Hong-Bay
    • Transactions on Electrical and Electronic Materials
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    • v.5 no.5
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    • pp.185-188
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    • 2004
  • Phase transitions from the amorphous to crystalline states, and vice versa, of GST(GeSbTe) and AST(AsSbTe) thin films by applying electrical pulses have been studied. These materials can be used as nonvolatile memory devices. The thickness of ternary chalcogenide thin films is approximately 100 nm. Upper and lower electrodes were made of AI. I-V characteristics after impressing the variable pulses to GST and AST films. Tc(crystallization temperature) of AST system is lower than that of the GST system, so that the current pulse width of crystallization process can be decreased.

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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Isolation of Myrosinase and Glutathione S-transferase Genes and Transformation of These Genes to Develop Phenylethylisothiocyanate Enriching Chinese Cabbage (배추에서 항암물질 phenylethylisothiocyanate의 다량 합성을 위한 myrosinase와 glutathione S-transferase 유전자 분리 및 이를 이용한 형질전환체 육성)

  • Park, Ji-Hyun;Lee, Su-Jin;Kim, Bo-Ryung;Woo, Eun-Teak;Lee, Ji-Sun;Han, Eun-Hyang;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.623-632
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    • 2011
  • To increase the anti-carcinogens phenylethylisothiocyanate (PEITC), myrosinase (MYR), and glutathione S-transferase (GST), genes related to PEITC pathway were isolated and the gene expressions were regulated by Agrobacterium transformation. Isolated cDNAs, MYR, and GST genes were 1,647 bp and 624 bp, respectively, and the protein expression was confirmed through pET system. Thereafter, we constructed a sense-oriented over-expressing myrosinase (pBMY) and RNAi down-regulated GST (pJJGST) binary vectors for the Chinese cabbage transformation. After the transformation, thirteen over-expressing transgenic Chinese cabbage plants (IMS) with pBMY and five down-regulated ones (IGA) with pJJGST were selected by PCR analysis. Selected $T_0$ transgenic plants were generated to $T_1$ plants by self-pollination. Based on the Southern blot analysis on these $T_1$ transgenic plants, 1-4 copies of T-DNA were transferred to Chinese cabbage genome. Thereafter, RNA expression level of myrosinase gene or GST gene was analyzed through real-time RT PCR of IMS, IGA, and non-transgenic inbred lines. In case of IMS lines, myrosinase gene was increased 1.03-4.25 fold and, in IGA lines, GST gene was decreased by 26.42-42.22 fold compared to non-transgenic ones, respectively. Analysis of PEITC concentrations using GC-MS it showed that some IMS lines and some IGA lines increased concentrations of PEITC up to 4.86 fold and up to 3.89 fold respectively compared to wild type. Finally in this study IMS 1, 3, 5, 12, and 15 and IGA 1, 2, and 4 were selected as developed transgenic lines with increasing quantities of anti-carcinogen PEITC.

Glutamate-rich 4 Binds to Kinesin Superfamily Protein 5A (Glutamate-rich 4와 kinesin superfamily protein 5A와의 결합)

  • Se Young Pyo;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Sang Jin Kim;Mooseong Kim;Jung Goo Lee;Dae-Hyun Seog
    • Journal of Life Science
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    • v.33 no.1
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    • pp.1-7
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    • 2023
  • Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.

Characterization of the Monoclonal Antibody Specific to Human S100A6 Protein (인체 S100A6 단백질에 특이한 단일클론 항체)

  • Kim, Jae Wha;Yoon, Sun Young;Joo, Joung-Hyuck;Kang, Ho Bum;Lee, Younghee;Choe, Yong-Kyung;Choe, In Seong
    • IMMUNE NETWORK
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    • v.2 no.3
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    • pp.175-181
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    • 2002
  • Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.

Analyzing Spatial and Temporal Variation of Ground Surface Temperature in Korea (국내 지면온도의 시공간적 변화 분석)

  • Koo Min-Ho;Song Yoon-Ho;Lee Jun-Hak
    • Economic and Environmental Geology
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    • v.39 no.3 s.178
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    • pp.255-268
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    • 2006
  • Recent 22-year (1981-2002) meteorological data of 58 Korea Meteorological Adminstration (KMA) station were analyzed to investigate spatial and temporal variation of surface air temperature (SAT) and ground surface temperature (GST) in Korea. Based on the KMA data, multiple linear regression (MLR) models, having two regression variables of latitude and altitude, were presented to predict mean surface air temperature (MSAT) and mean ground surface temperature (MGST). Both models showed a high accuracy of prediction with $R^2$ values of 0.92 and 0.94, respectively. The prediction of MGST is particularly important in the areas of geothermal energy utilization, since it is a critical parameter of input for designing the ground source heat pump system. Thus, due to a good performance of the MGST regression model, it is expected that the model can be a useful tool for preliminary evaluation of MGST in the area of interest with no reliable data. By a simple linear regression, temporal variation of SAT was analyzed to examine long-term increase of SAT due to the global warming and the urbanization effect. All of the KMA stations except one showed an increasing trend of SAT with a range between 0.005 and $0.088^{\circ}C/yr$ and a mean of $0.043^{\circ}C/yr$. In terms of meteorological factors controlling variation of GST, the effects of solar radiation, terrestrial radiation, precipitation, and snow cover were also discussed based on quantitative and qualitative analysis of the meteorological data.