• Title/Summary/Keyword: glutathione transferase

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Shotgun Phage Display of Lactobacillus casei BL23 Against Collagen and Fibronectin

  • Munoz-Provencio, Diego;Monedero, Vicente
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.197-203
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    • 2011
  • Lactobacilli are normal constituents of the intestinal microbiota, and some strains show the capacity to bind to extracellular matrix proteins and components of the mucosal layer, which represents an adaptation to persist in this niche. A shotgun phage-display library of Lactobacillus casei BL23 was constructed and screened for peptides able to bind to fibronectin and collagen. Clones showing binding to these proteins were isolated, which encoded overlapping fragments of a putative transcriptional regulator (LCABL_29260), a hypothetical protein exclusively found in the L. casei/rhamnosus group (LCABL_01820), and a putative phage-related endolysin (LCABL_13470). The construction of different glutathione S-transferase (GST) fusions confirmed the binding activity and demonstrated that the three identified proteins could interact with fibronectin, fibrinogen, and collagen. The results illustrate the utility of phage display for the isolation of putative adhesins in lactobacilli. However, it remains to be determined whether the primary function of these proteins actually is adhesion to mucosal surfaces.

Effects of Protein and Fiber on Antioxidant Enzyme Activites of Brain in Ethanol-Treated Rats (에탄올을 투여한 흰쥐 노조직의 항산화효소계 활성에 미치는 단백질과 섬유소의 영향)

  • 이미경
    • Journal of Nutrition and Health
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    • v.33 no.6
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    • pp.613-618
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    • 2000
  • This study was to investigate the effect of dietary protein and fiber on the antioxidant enzyme activities of brain in acute or chronic ethanol-treated rats. Male Sprague-Dawley rats were fed on diets containing two levels of protein(7%, 20%) with two levels of fiber (5%, 10%) Rats were administered 40%(v/v) ethanol(5g/kg body weight)orally 90min before decaptiation in acute ethanol-treated groups and 25%(v/v) ethanol(5g/kg body weight) once a day for 5 weeks in chronic ethanol treated-groups. The rats were sacrificed after 5 weeks of feeding periods. Superoxide dismutase and gluthathione S-transferase activities were lower in chronic ethanol-treated groups than acute ethanol-treated groups whereas catalase and glutathuone peroxidase activities were significantly increased by chronic ethanol treatment. Low protein supplement accelerated to change of their activities however dietary fiber levels did not affect antioxidant enzyme activities. Chronic ethanol treatment and/or low protein supplement results in increasing the brain lipid peroxide content but in lowering glutathione level. (Korean J Nutrition 33(6) ; 613~618, 2000)

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대장균내에서 발현된 돼지 TGF-$\beta$1의 분리 및 면역학적 항원성 보유검증

  • Choi, Eun Young;;Kim, Pyung Hyun;Byeon, Woo-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.137-143
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    • 1997
  • Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

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Effect of Nonlamellar-Prone Lipids on Protein Encapsulation in Liposomes

  • Ahn, Tae-Ho;Chi, Youn-Tae;Yun, Chul-Ho
    • Macromolecular Research
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    • v.17 no.12
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    • pp.956-962
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    • 2009
  • We investigated the effect of two nonlamellar-prone lipids, phosphatidylethanolamine (PE) and dioleoylglycerol (DOG), on the efficiency of protein encapsulation in liposomes. When the phosphatidylcholine (PC) matrix was replaced with PE or DOG during liposome formulation, the amounts of glutathione S-transferase and bovine serum albumin entrapped in the vesicles increased with increasing PE or DOG concentration. The presence of PE and DOG synergistically affected protein entrapment. These results suggest that protein encapsulation can be enhanced by the presence of nonlamellar lipids and/or lipid-induced membrane properties.

PHARMACOGENOMICS IN RELATION TO TAILOR-MADE DRUGS -INTRODUCTION-

  • Satoh, Tetsuo
    • 한국약용작물학회:학술대회논문집
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    • 2006.11a
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    • pp.51-66
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    • 2006
  • The field of cytochrome P450 pharmacogenomics has progressed rapidly during the past 25 years. Recently, conjugating enzymes including sulfotransferase, acetyltransferase, glucuronosyltransferase and glutathione transferase have been also extensively studied. All the major human drug-metabolizing P450 enzymes and some conjugating enzymes have been identified and cloned, and the major gene variants that cause inter-individual variability in drug response and are related to adverse drug reactions have been identified. This information now provides the basis for the use of predictive pharmacogenomics to yield drug therapies that are more efficient and safer. Today, we understand which drugs warrant dosing based on pharmacogenomics to improve drug treatment. It is anticipated that genotyping could be used to personalize drug treatment for vast numbers of subjects, decreasing the cost of drug treatment and increasing the efficacy of drugs and health in general. It is assumed that such personalized P450 gene-based treatment which is so-called tailor(order)-made drug therapy would be relevant for 10-20% of all drug therapy in the future.

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Effects of Carcinogens and Partial Hepatectomy on the Nitrogen Utilizing and the Xenobiotic Metabolizing Enzymes in the Hepatic Tissues of Rats (발암원과 부분간절제술 처리에 의한 백서 간 조직중 질소이용계 및 이물질 대사계 효소의 변화)

  • 박상철;김응국;곽상준;이건욱;김수태
    • Environmental Mutagens and Carcinogens
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    • v.8 no.1
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    • pp.22-34
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    • 1988
  • The biochemical changes of the hepatic tissues, induced by the carcinogen treatment such as diethylnitrosamine and acetamidofluorene in combination with the partial hepatectomy after Solt and Farber, were determined for the characterization of the induction of the proliferative capacity and the environmental adaptability of the carcinogenic tissues during the malignant transformation process. For the study of the proliferative capacity of the tissues, the activities of the enzymes, related with the nitrogen trapping mechanism, such as glutamine synthetase and gamma-glutamyltranspeptidase, were monitroed, while the cintents of cytochrome P450's and their isozymic patterns as well as the activities of the glutathione S-transferase were determined in the function of time after the hepatocarcinogenic stimuli.

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Estimation of Death Time by Changes of Postmortem Xanthine Oxidase Activity in Rats

  • Yoon, Hyung-Won;Yoon, Chong-Guk;Cho, Hyun-Gug
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.439-442
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    • 2006
  • To evaluate the postmortem changes in activities of oxygen free radical metabolizing enzymes, the rats were sacrificed with cervical dislocation and were kept in an incubator at $25^{\circ}C$, 70% of humidity for 12 hours. The activities of aniline hydroxylase, catalase, glutathione-S-transferase and superoxlde dismutase were decreased with the time. On the other hand, the activity and type conversion ratio (type D ${\to}$type O) of hepatic xanthine oxidase (XO) were gradually increased. From these changes of XO, the estimation of death time (mathematical equation) could be determined with the least square method. To clarify the cause of increasing XO activity, enzyme kinetics were examined. The Km values of XO were decreased with the time. In conclusion, the determination of liver XO activity might be used for the estimation of death time in the early postmortem period.

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Anti-HIV and Antihepatotoxic Constituents from Medicinal Plant Resources

  • Park, Jong-Cheol;Park, Ju-Gwon;Hur, Jong-Moon;Hwang, Young-Hee;Jung, Deuk-Young
    • Plant Resources
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    • v.4 no.3
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    • pp.196-199
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    • 2001
  • Medicinal plants were screened for the inhibitory effects on human immunodeficiency virus type 1 pretense. Of the extracts tested, the strong inhibitory effects were observed in the acetone extracts of the pericarp of Camellia japonica. Camelliatannin H from the pericarp of C. japonica showed a potent inhibitory activity on HIV-1 pretense. Effects of the extract and compound from leaves of Zanthoxylum piperitum on the enzyme activities were investigated in the liver of bromobenzene-treated rats. The methanol extract and protocatechuic acid isolated from Z. pipetitum reduced the activity of aniline hydroxylase that increased by bromobenzene, while did not affect the activities of aminopyrin N-demethylase and glutathione S-transferase. The extract and protocatechuic acid recovered significantly the activity of epoxide hydrolase decreased by bromobenzene.

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Effects of Vinorelbine on Cisplatin Resistance Reversal in Human Lung Cancer A549/DDP Cells

  • Zhou, Yu-Ting;Li, Kun;Tian, Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4635-4639
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    • 2013
  • Multi-drug resistance (MDR) is an essential aspect of human lung cancer chemotherapy failure. Recent studies have shown that vinorelbine is involved in underlying processes in human tumors, reversing the MDR inseveral types of cancer cells. However, the roles and potential mechanism are not fully clear. In this study, we explored effects of vinorelbine in multi-drug resistance reversal of human lung cancer A549/DDP cells. We found that vinorelbine increased drug sensitivity to cisplatin and intracellular accumulation of rhodamine-123, while decreasing expression of P-glycoprotein (P-gp), multi-drug resistance-associated protein (MRP1) and glutathione-S-transferase ${\pi}$ (GST-${\pi}$) in A549/DDP cells. At the same time, we also established downregulation of p-Akt and decreased transcriptional activation of NF-${\kappa}B$ and twist after vinorelbine treatment. The results indicated that vinorelbine might be used as a potential therapeutic strategy in human lung cancer.

Mutation of Cysteine-115 to Alanine in Nicotiana glutinosa Ornithine Decarboxylase Reduces Enzyme Activity

  • Lee, Yong-Sun;Cho, Young-Dong
    • BMB Reports
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    • v.34 no.5
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    • pp.472-477
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    • 2001
  • Ornithine decarboxylase (ODC, EC 4.1.1.17) is the first and key enzyme in eukaryotic polyamine biosynthesis. The cDNA encoding ornithine decarboxylase from Nicotiana glutinosa was cloned ($GeBank^{TM}$ AF 323910) and expressed in E. coli. Site directed mutagenesis were performed on several highly conserved cysteine residues. Among the mutants, C115A showed significant changes in the kinetic properties. The $K_m$ value of the C115A mutant was $1790\;{\mu}M$, which was 3-fold higher than that of the wild-type ODC. There was a dramatic decrease in the $k_{cat}$, values of the C115A mutant, compared to that of the wild-type ODC, which had a $k_{cat}$ value of $77.75\;s^{-1}$. C115A caused a shift in the optimal pH from 8.0 to 8.4. Considering these results, we suggest that cys-115 is involved in the catalytic activity of N. glutinosa ODC.

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