• Title/Summary/Keyword: glutathione transferase

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Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Expression of Recombinant Hybrid Peptide Gaegurin4 and LL37 using Fusion Protein in E. coli (Glutathione S-Transferase에 융합한 재조합 Hybrid Peptide Gaegurin-LL37의 대장균에서의 발현)

  • Bayarbat, Ishvaanjil;Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.92-97
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    • 2012
  • Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1-16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

Effect of Insecticide Carbofuran and Phenobarbital Sodium and 3-Methylcholanthrene on Activity of Enzyme in Israeli Carp(Cyprinus israeli carpio L.) (살충제 Carbofuran과 Phenobarbital Sodium 및 3-Methylcholanthrene이 이스라엘 잉어의 효소활성에 미치는 영향)

  • Rim, Yo-Sup;Jeong, Jae-Hun;Han, Seong-Soo
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.77-83
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    • 1996
  • Effects of insecticide carbofuran and Phenobarbital sodium(PB) or 3-methylcholanthrene(3-MC) on activities of several enzymes in israeli carps were investigated. Survival number of Israeli carp was the same as that of control when PB and 3-MC only was treated, individually and that was low compared to control when carbofuran only was treated. But survival rate of Israeli carp was high compared to individual treatment of carbofuran when combination treatment of carbofuran and PB or 3-MC was carried out. These results indicate that PB and 3-MC can intervene to detoxify carbofuran exposed to israeli carp. In in vivo test for the effect of this chemicals on activity of enzyme in israeli carp, activities of acetylcholinesterase(AChE) and glutathione S-transferase(GST) were inhibited in carbofuran treatment, but did not in combination treatment of carbofuran and P3 or 3-MC. Activities of UDP-glucuronosyltransfe-rase (UDPGT) and cytochrome P-450-dependent monooxygenase increased in individual or combined treatments of carbofuran and PB or 3-MC. These results suggest that a simultaneous application of carbofuran and PB or 3-MC is critical for the enhancement of activity of AChE, GST, UDPGT and monooxygenase and the protection of Israeli carp from carbofuran toxicity.

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Detoxification Effect of Microcluster-Water on Bromobenzene-Induced Liver Damaged Mice (Bromobenzene으로 유도된 간 손상 마우스에 대한 Microcluster수의 효과)

  • Park, Bum-Ho;Back, Kyung-Yern;Lee, Sang-Il;Kim, Soon-Dong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.3
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    • pp.291-297
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    • 2007
  • This study was conducted to investigate the hepatic detoxification effect of microcluster-water (McW). Animal experiments were divided into 4 groups: distilled water intake group (DC), distilled water intake-bromobenzene treated group (DB), McW intake group (MC), and McW intake-bromobenzene treated group (MB). There were no significant differences in alanine aminotransferase and aspartate aminotransferase activities between DC and MC groups, but the activities in MB group were significantly (p<0.05) lower than those in DB group. No apparent changes of aniline hydrolase activity were shown in all experimental groups, while glutathione S-transferase activity in MC and MB groups was higher than that in DC and DB, respectively. The content of hepatic lipid peroxide in DC group was similar to that of MC group. In addition, the contents in DB and MB groups were significantly (p<0.05) increased than that of DC group. The increasing rate in MB group was lower than that of DB group. Also, the electron donating activity of McW was significantly (p<0.05) higher than that of distilled water. From these results, it could be suggested that McW has the possibility of having detoxification effect of bromobenzene induced hepatic injury by increasing glutathione S-transferase, which is known as a kind of hepatiic detoxification enzyme.

Lack of Any Association of GST Genetic Polymorphisms with Susceptibility to Ovarian Cancer - a Meta-analysis

  • Han, Li-Yuan;Liu, Kui;Lin, Xia-Lu;Zou, Bao-Bo;Zhao, Jin-Shun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6131-6136
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    • 2014
  • Objective: Epidemiology studies have reported conflicting results between glutathione S-transferase Mu-1 (GSTM1), glutathione S-transferase theta-1 (GSTT1) and glutathione S-transferase pi-1 (GSTP1) and ovarian cancer (OC) susceptibility. In this study, an updated meta-analysis was applied to determine whether the deletion of GSTM1, GSTT1 and GSTP1 has an influence on OC susceptibility. Methods: A published literature search was performed through PubMed, Embase, Cochrane Library, and Science Citation Index Expanded database for articles published in English. Pooled odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated using random or fixed effects models. Heterogeneity between studies was assessed using the Cochrane Q test and $I^2$ statistics. Sub-group analysis was conducted to explore the sources of heterogeneity. Sensitivity analysis was employed to evaluate the respective influence of each study on the overall estimate. Results: In total, 10 published studies were included in the final analysis. The combined analysis revealed that there was no significant association between GSTM1 null genotype and OC risk (OR=1.01, 95%CI: 0.91-1.12). Additionally, there was no significant association between GSTT1 genetic polymorphisms and OC risk (OR=0.98, 95% CI: 0.85-1.13). Similalry, no significant associations were found concerning the GSTP1 rs1695 locus and OC risk. Meanwhile, subgroup analysis did not show a significant increase in eligible studies with low heterogeneity. However, sensitivity analysis, publication bias and cumulative analysis demonstrated the reliability and stability of the current meta-analysis. Conclusions: These findings suggest that GSTs genetic polymorphisms may not contribute to OC susceptibility. Large epidemiological studies with the combination of GSTM1 null, GSTT1 null and GSTP1 Ile105Val polymorphisms and more specific histological subtypes of OC are needed to prove our findings.

Association Study of Glutathione-S-Transferase M1/T1 Gene Polymorphism with Deficiency-Excess Differentiation-syndrome in Korean Bronchial Asthmatics (한국인 기관지 천식 환자에서 허설변증과 Glutathione-S-Transferase 유전자의 다형성 연구)

  • Yu, Seung-Ryeol;Jeong, Seung-Yeon;Jung, Ju-Ho;Kim, Jin-Ju;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.28 no.3
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    • pp.453-463
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    • 2007
  • Backgrounds : Glutathione-s-transferase (GST) is a kind of phase II metabolism enzyme and plays an important role in the detoxification of various toxic chemicals. It was reported that the genetic polymorphism of GSTM1 and GSTT1 genes may be responsible for asthma development and susceptibility to allergy. Traditional oriental medicine uses a unique diagnostic technique. differentiation-syndrome. to analyze signs and symptoms of patients synthetically. Through differentiation-syndrome. asthma patients can be divided into two groups: the deficiency syndrome group (DSG) and the excess syndrome group (ESG). Objectives : The purpose of this study was to investigate the possible association of GST gene polymorphism with clinical phenotype by differentiation-syndrome of bronchial asthma patients. Materials and Methods : One hundred and ten participants were evaluated by pulmonary function test. Patients with 53 DSG and 31 ESG by differentiation-syndrome were assessed for genetic analysis. GSTM1 and GSTT1 deletion polymorphism was performed by polymerase chain reaction (PCR). Results : GSTM1 gene deletion was detected in 43.4% of individuals in the DSG and in 38.71 % in the ESG. The distribution of GSTM1 polymorphism between DSG and ESG was not significantly different [$x^2$=0.1767, p=0.6742; OR(95% CI)=1.2139(0.4915-2.9979)]. The proportion of GSTT1 null genotypes was 41.51% in the DGS and 45.16% in the ESG. The distribution of GSTT1 polymorphism between DSG and ESG was also not significantly different [$x^2$=0.1065, p=0.7442; OR(95% CI)=0.8618(0.3525-2.1065)]. In the combined analysis of GSTM1 and GSTT1 genes, the frequency of both null type of GSTM1/GSTT1 genes was not significantly different from both positive type of GSTM1/GSTT1 genes[$x^2$=0.0768, p=0.7817; OR(95% CI)=1.2000(0.3303-4.3602)] Conclusions : These results indicate that polymorphism of the GST gene might not be associated with the symptomatic classification of DSG and ESG by differentiation-syndrome in Korean asthmatics.

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The Anticancer Effects of Doenjang Made with Various Kinds of Salt (소금 종류를 달리하여 제조한 된장에서의 항암효과)

  • Lee, Kyeoung-Im;Park, Kun-Young;Ahn, Hyung-Ki
    • Culinary science and hospitality research
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    • v.17 no.5
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    • pp.241-252
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    • 2011
  • The study was carried out to evaluate the anticancer effects in Korean soybean paste(doenjang) added with sun-dried salt(S-D), roasted salt(R-D), bamboo salt roasted once(B1-D) or bamboo salt roasted nine times(B9-D). In MTT assay, S-D, R-D, B1-D and B9-D exhibited a significant inhibitory activity(64-87% and 68-92%) against the proliferation of AGS human gastric adenocarcinoma cells by adding 0.05% and 0.1% of methanol extract. Among Balb/c mice injected with sarcoma-180 cells, the bodies, livers, spleens, kidneys and heart weight of the mice administered with 4 kinds of doenjang extracts were heavier than the non-administered ones of the group. However, no difference was found between the control and doenjang administered groups. Four kinds of doenjang inhibited significantly the tumor growth, especially R-D and B1-D showing an inhibition of tumor cell growth up to 97% by the administration of 1.0 mg/kg methanol extracts of doenjang. The activity of natural killer(NK) cells was relatively high in mice administered with four kinds of doenjang. Particularly, mice administered with R-D methanol extract showed a stronger activity of 88.2%. The activity of glutathione S-transferase(GST) in mice administered with four kinds of doenjang was higher than that of the non-administered group. In particular, the GST activity was the strongest in the group with B1-D. As these results indicate the various kinds of salt have different effects on the anticancer activity of doenjang.

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Protective Effect of Whagan-Jeon (huaganjian) on Acetaminophen-induced Hepatotoxicity (화간전이 아세트아미노펜에 의한 간독성에 미치는 영향)

  • 박철수;김기열;이채중;안중환;김종대;남경수
    • The Journal of Korean Medicine
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    • v.23 no.3
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    • pp.33-42
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    • 2002
  • Objective : This study was performed to investigate the activity of Whagan-Jeon (huaganjian) in protection against acetaminophen (AAP)-induced hepatotoxicity and the possible mechanisms in vivo. Methods : The following were performed : Serum ALT, depletion of hepatic glutathione (GSH) levels, the microsomal p. nitrophenol hydroxylation activity, microsomal aniline hydroxylation activity, genomic DNA fragmentation and its reversal, hepatic glutathione-S-transferase (GST) activity, and hepatic NAD(P)H:quinone oxidoreductase (QR) activity Results : Whagan-Jeon (huaganjian) protected against AAP-inducedhepatotoxicity by the increase of GSH levels, inhibition of P450 2E1-specific metabolic activities, attenuation of hepatic DNA damage, and induction of GST and QR activities in vivo. Conclusions : In conclusion, Whagan-Jeon (huaganjian) was effective in protection against AAP-induced hepatoxicity.

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The Effects of NOG-YONG Aqua-Aqupuncture Solution on Cytotoxicities in Primary Culture of Adult Rat Hepatocytes

  • Byun, Boo-Hyeong;Jeong, Hye-Kwang
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 2002.05a
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    • pp.147-147
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    • 2002
  • This experimental study was carried out to investigate on the safety of Nog-yong aqua-acupuncture solution in primary culture of adult rat hepatocytes. Primary culture of adult rat hepatocytes has been considered as a ideal model for toxicological studies because cultured hepatocytes maintained many liver-specific functions. In this research, we investigated the effects of Nog-yong aqua-aqupunture(1-10$\mu\textrm{g}$/ml) on cytotoxicites in primary culture of adult rat hepatocytes using LDH release assays. Hepatic glutathione level. glutathione-S-transferase activity, and albumin synthesis were not affected by treatment with Nog-yong aqua-aqupuntur alone. Nog-yong aqua-aqupuncoure solution(0.5-$10{\mu}\textrm{g}$/ml) on cytotoxicites in primary culture of adult rat hepatocytes using LDH release not significantly affected normal functional charaterists.

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Cutaneous Toxicity of Xylene Application to Rat Skin

  • Jeon, Tae-Won;Lee, Sang-Hee;Yoon, Chong-Guk
    • Biomedical Science Letters
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    • v.8 no.2
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    • pp.83-88
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    • 2002
  • To investigate the skin toxicity of xylene, xylene (25 mg/$\textrm{cm}^2$) has been sequentially applied to the rat skin for four days. On the light microscopic examination, epithelium was left out with infiltration of inflammatory cells in border with dermis, and formation of new epithelial layer was shown under the inflammatory zone. Application of xylene to the rat skin showed the marked rise of cutaneous xanthine oxidase activity whereas, He activities of oxygen free radical scavenging enzymes, superoxide dismutase and glutathione S-transferase, were significantly declined. Furthermore, the content of cutaneous glutathione was more and less decreased in rat skin applied with xylene. In conclusion, these results suggest that a part of oxygen free radical may be responsible for morphological changes in skin by applying xylene to the rat skin.

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