• Title/Summary/Keyword: S-transferase

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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.

Lymphocyte DNA Damage and Anti-Oxidative Parameters are Affected by the Glutathione S-Transferase (GST) M1 and T1 Polymorphism and Smoking Status in Korean Young Adults (흡연 여부에 따른 Glutathione S-transferase (GST) M1 및 T1 유전자 다형성이 우리나라 젊은 성인의 임파구 DNA 손상과 항산화 영양상태 지표들 간의 관련성에 미치는 영향)

  • Han, Jeong-Hwa;Lee, Hye-Jin;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.366-377
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    • 2011
  • Glutathione S-transferase (GST) is a multigene family of phase II detoxifying enzymes that metabolize a wide range of exogenous and endogenous electrophilic compounds. GSTM1 and GSTT1 gene polymorphisms may account for inter-individual variability in coping with oxidative stress. We investigated the relationships between the level of lymphocyte DNA and antioxidative parameters and the effect on GST genotypes. GSTM1 and GSTT1 were characterized in 301 young healthy Korean adults and compared with oxidative stress parameters such as the level of lymphocyte DNA, plasma antioxidant vitamins, and erythrocyte antioxidant enzymes in smokers and non smokers. GST genotype, degree of DNA damage in lymphocytes, erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px), and plasma concentrations of total radical-trapping antioxidant potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene, and cryptoxanthin were analyzed. Lymphocyte DNA damage assessed by the comet assay was higher in smokers than that in non-smokers, but the levels of plasma vitamin C, ${\beta}$-carotene, TRAP, erythrocyte catalase, and GSH-Px were lower than those of non-smokers (p < 0.05). Lymphocyte DNA damage was higher in subjects with the GSTM1- or GSTT1-present genotype than those with the GSTM1-present or GSTT1- genotype. No difference in erythrocyte antioxidant enzyme activities, plasma TRAP, or vitamin levels was observed in subjects with the GSTM1 or GSTT1 genotypes, except ${\beta}$-carotene. Significant negative correlations were observed between lymphocyte DNA damage and plasma levels of TRAP and erythrocyte activities of catalase and GSH-Px after adjusting for smoking pack-years. Negative correlations were observed between plasma vitamin C and lymphocyte DNA damage only in individuals with the GSTM1-present or GSTT1- genotype. The interesting finding was the significant positive correlations between lymphocyte DNA damage and plasma levels of ${\alpha}$-carotene, ${\beta}$-carotene, and cryptoxanthin. In conclusion, the GSTM1- and GSTT1-present genotypes as well as smoking aggravated antioxidant status through lymphocyte DNA damage. This finding confirms that GST polymorphisms could be important determinants of antioxidant status in young smoking and non-smoking adults. Consequently, the protective effect of supplemental antioxidants on DNA damage in individuals carrying the GSTM1- or GSTT1-present genotypes might show significantly higher values than expected.

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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Chemopreventive activity of Prunella Herba Vulgaris L. Aqua-acupuncture Solution (댑싸리하고초(夏枯草) 약침액(藥鍼液)의 암예방 활성)

  • Park Shin-Hwa;Lim Jong-Kook
    • Korean Journal of Acupuncture
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    • v.18 no.1
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    • pp.11-20
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    • 2001
  • Cancer chemoprevention refer to the use of natural or synthetic substances to prevent the initiational and promtional events that occur during the process of carcinogenesis. The effect of Prunella Herba Vulgaris L. Aqua-acupuncture Solution (PVAS) and Prunella Herba Vulgaris L. Water-extracted Solution (PVWS) on the induction of phase II detoxification enzyme (quinone reductase, Glutathione S-transferase) and inhibition of phase I enzyme (cytochrome P4501A1) and benzo[a]pyrene-DNA adduct formation was examined. PVAS is potent inducers of quinone reductase activity. Glutathione levels were increased with PVAS, in cultured murine hepatoma Hepa1c1c7 cells. In addition glutathione S-transferase levels were increased with PVAS. However, there was 45.2% inhibition in the activity of cytochrome P4501A1 enzyme with the treatment of PVAS, $5{\times}$. At concentration of $1{\times}$ and $3{\times}$ of PVAS, the binding of $[^3H]B[a]P$ metabolites to DNA of NCTC-clone 1469 cell was inhibited by 25.3%, 45.0%, respectively. These results suggest that PVAS has chemopreventive potential by inducing quinone reductase and glutathione S-transferase activities, increasing GSH levels, inhibiting the activity of cytochrome P4501A1 and benzo[a]pyrene-DNA adduct formation.

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Effect of Carbon Tetrachloride on the Changes of Xanthine Oxidase Activity in Rate Previously Fed Low or High Protein Diet (식이성 단백질 함량에 따른 흰쥐에 사염화탄소 투여가 Xanthine Oxidase활성에 미치는 영향)

  • 윤종국;이상일;신중규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.6
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    • pp.527-537
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    • 1991
  • To evaluate an effect of liver xanthine oxidase on the induction of liver damage, carbon tetrachloride (CCl4) was intraperitoneally injected twice at 0.1ml/100g body weight to the rate fed a low (LP)or high protein diet(HP) while the control group fed LP or HP received only olive oil. The changing rate of liver xanthine oxidas activity was compared with that of a free radical generating enzyme, liver aniline hydroxylase and a scavenging enzyme, glutathions S-transferase activity between the rate fed a LP and those fed HP, and the two groups treated with CCl4. Concomitantly, the degree of liver damage which could be considered as the paramete for CCl4 metabolism in case of CCl4-intoxicated animal was observed in the present experimental conditions and the effect of allopurinol, xanthine oxidase inhibitor, on the CCl4-toxicity of rate liver was alos demostrated. On the other hand, the comparative effect of actinomycin D on the liver and serum xanthine oxidase of CCl4-treated rats fed HP with that of those fed LP and the kinetics of purifed liver enzyme from the liver of CCl4-treated rats fed HP was also compared with that of those fed LP to clarify the differences of xanthine oxidase activity between two groups. The increasing rate of liver weigth/body wt, serum levels of ALT and the decreasing rate of hepatic ALT activity and protein contents to each control group were higher in CCl4-treated rats fed HP than those fed LP. Under the animal models as indentified by the present data herein, the liver xanthine oxidase activity was higher in CCl4-treated rats fed HP than those fed LP, and the control group fed HP also showed the much higher activity xanthine oxidase than that fed LP, whereas there were no differences in the activity of hepatic aniline hydroxylase and glutathions S-transferase between the two group treated with CCl4. Although the hepatic aniline hydroxylase activity was somewhat higher in the rats fed HP than those fed LP, the increasing rate of liver xanthine oxidase to the rats fed LP was higher in those fed HP than that of liver aniline hydroxylase. The degree of liver damage identified such as liver weight and serum ALT activity was less in the CCl4-treated rats pretreated with allopurinol. These results suggest that even a system at which xanthine oxidase acts as well as the drug metabolizing enzyme may influence the acelatin of CCl4 metabolism. In addition, the purified liver xanthine oxidase from CCl4-treated rats fed HP showed decreased Km value when compared to its control group. The Km value of liver xanthine oxidase of CCl4-treated rats fed LP showed a similar Km value with its control group. Furthermore, the decreasing rate of liver and serum xanthine oxidase acitivity in CCl4-treated rats pretreated with actinomycin D to the CCl4-treated rats was higher in rats fed HP than in those fed LP. These results suggest that the inductino of xanthine oxidase in CCl4-treated rats fed HP may be greater than in those fed LP.

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Effect of Aging on the Liver Damage in Bromobenzene-pretreated Rats (연령이 다른 흰쥐에 Bromobenzene 미치는 영향)

  • 한선일;윤형원;윤종국
    • Biomedical Science Letters
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    • v.5 no.2
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    • pp.201-208
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    • 1999
  • To evaluate an effect of growth periods on the bromobenzene-induced liver damage, bromobenzene was administrated to 5-week-old rats and 10-week-old rats pretreated with bromobenzene 5 times every other day for 10 days and then the animals were sacrificed. The results were obtained as follows; The increasing rate of serum levels of alanine aminotransferase, xanthine oxidase activity, hepatic lipid peroxide contents, liver weight per body weight (%) and decreasing rate of hepatic contents of protein to each control group were higher in 10-week-old rats than 5-week-old rats by the pretreatment of bromobenzene. According to the above results, 10-week-old rats indicated more severe liver injury than 5-week-old those in case of bromobenzene pretreatment. On the other hand, hepatic aniline hydroxylase activity was more increased in 10-week-old rats than 5-week-old rats both in control and bromobnezene pretreated rats where as the reverse in hepatic glutathione S-transferase. In case of hepatic GSH determination at the intervals of 2, 4, 8, 24 hours throughout 24 hr after administration of single dose of bromobenzene to 5-week-old and 10-week-old rats both in control and bromobnezene pretreated, the rate of GSH utilization was lower in 10-week-old rats than 5-week-old rats. In conclusion, from the above experimental, it is deduce that the 10-week-old rats showed more severe liver injury than 5-week-old rats by the bromobenzene treatment because the disposal ability of bromobenzene in liver was lower in 10-week-old rats than 5-week-old rats.

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Effect of Tota1 Saponin from Red Ginseng on Acvtivities of Antioxidant Enzymes in Pregnant Rats (홍삼 사포닌이 수태중인 흰쥐의 항산화 효소활성에 미치는 영향)

  • Song, Yong-Bum;Kwak, Yi-Seong;Park, Ki-Hyun;Chang, Sung-Keun
    • Journal of Ginseng Research
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    • v.26 no.3
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    • pp.139-144
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    • 2002
  • Pregnancy is a physiological state accompained by a high energy demand of many bodily functions and an increased oxygen requirement. Because of the increased intake and utilization of oxygen, increased levels of oxidative stress would be expected. So we observed the activities of the hepatic antioxidant enzymes from rat treated with total saponin from the red ginseng against free raicals produced in pregnant rats. The activity of superoxide dismutase (SOD) in the control group was slightly decreased during pregnancy, and SOD activity in total saponin treated group was not observed any siginificant change compared with the control group. The activities of glutathione peroxidase (GPX), glutathione reductase (GRD) and catalase in the control group have shown the decreasing tendency during pregnancy, whereas the activities of GRD and catalase in total saponin treated group showed significant increased tendency compared with the control group. GPX activity in total saponin treated group was slightly decreased tendnency compared with the control group. The activity of glutathione-S-transferase (GST) in the control group was increased to keep the state of homaeostasis tendency in pregnant rats. On the other hand, the activity of GST after total saponin treatment was increased than control group. Activity of all enzymes in the control group and total saponin treated group recovered the normal level after delivery of rats. In spite of the physiological changes in vivo, the inflaunce of total saponin on activaties of hepatic antioxidant enzyme in pregnant rats seems to be regulated the biological homeostatic adaptation mechanism which protects the maternal liver aganist oxygen induced toxicity