• Title/Summary/Keyword: $\gamma$-glutamylcysteine

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Regulation of the Gene Encoding Glutathione Synthetase from the Fission Yeast

  • Kim, Su-Jung;Shin, Youn-Hee;Kim, Kyung-Hoon;Park, Eun-Hee;Sa, Jae-Hoon;Lim, Chang-Jin
    • BMB Reports
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    • v.36 no.3
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    • pp.326-331
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    • 2003
  • The fission yeast cells that contained the cloned glutathione synthetase (GS) gene showed 1.4-fold higher glutathione (GSB) content and 1.9-fold higher GS activity than the cells without the cloned GS gene. Interestingly, $\gamma$-glutamylcysteine synthetase activity increased 2.1-fold in the S. pombe cells that contained the cloned GS gene. The S. pombe cells that harbored the multi copy-number plasmid pRGS49 (containing the cloned GS gene) showed a higher level of survival on solid media with cadmium chloride (1 mM) or mercuric chloride ($10\;{\mu}M$) than the cells that harbored the YEp357R vector. The 506 bp upstream sequence from the translational initiation point and N-terminal8 amino acid-coding region were fused into the promoteriess $\beta$-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pUGS39. Synthesis of $\beta$-galactosidase from the fusion plasmid pUGS39 was significantly enhanced by cadmium chloride and NO-generating S-nitroso-N-acetylpenicillamine (SNAP) and sodium nitroprusside (SN). It was also induced by L-buthionine-(S,R)-sulfoximine, a specific inhibitor of $\gamma$-glutamylcysteine synthetase (GCS). We also found that the expression of the S. pombe GS gene is regulated by the Atf1-Spc1-Wis1 signal pathway.

Construction of the recombinant yeast strain with transformation of rice starch-saccharification enzymes and its alcohol fermentation (유전자 형질전환을 통한 쌀 전분 분해효소 재조합 효모균주의 개발과 발효특성조사)

  • Lee, Ja-Yeon;Chin, Jong-Eon;Bai, Suk
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.220-225
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    • 2016
  • To improve antioxidant glutathione (GSH) content and saccharification ability in sake yeasts of Saccharomyces cerevisiae, the ${\gamma}$-glutamylcysteine synthetase gene (GSH1) from S. cerevisiae, glucoamylase gene (GAM1) and ${\alpha}$-amylase gene (AMY) from Debaryomyces occidentalis were co-expressed in sake yeasts for manufacturing a refreshing alcoholic beverage abundant in GSH from rice starch. The extracellular GSH content of the recombinant sake yeasts increased 1.5-fold relative to the parental wide-type strain. The saccharification ability by glucoamylase of the new yeast strain expressing both GAM1 and AMY genes was 2-fold higher than that of the yeast strain expressing only GAM1 gene when grown in the culture medium containing 2% (w/v) rice starch. It generated 11% (v/v) ethanol from 20% (w/v) rice starch and consumed up to 90% of the starch content after 7 days of fermentation.

Bromate Formation by Ozonation Process and It′s Effect on Renal Toxicity in rat (오존처리에 의한 Bromate의 생성 및 흰쥐의 신장독성에 미치는 영향)

  • 정운용;이무강;최종원
    • Journal of Life Science
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    • v.12 no.4
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    • pp.442-451
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    • 2002
  • In oder to investigate the effects of pH and temperature on the formation of bromate ion, which is ozonation by-products of bromine containing natural water. At the same intial pH condition, the increase of pH shown similar trends even if the reaction variables such as temperature and reaction time of ozonation were changed. As pH and temperature were increasing, the bromate concentration was increased but bromine components (HOBr/OBr-) were decreased with increasing pH from 3 to 10. Lipid peroxide content in the kidney was increased by bromate which was ingestion with 0.4g/L for 24 weeks in drinking water. Renal cytosolic enzyme system (XO, AO) of bromate group were significantly increased in comparison with those of normal group. But microsomal enzyme system were not affected. BUN level and urinary ${\gamma}$-glutamyltransferase activity were significantly increased in comparison with those of the normal. But, urinary lactate dehydrogenase activity was not affected. Renal glutathione content of rat was significantly decreased in comparison with those of normal rat given bromate. Renal glutathione S-transferase and ${\gamma}$-glutamylcysteine synthetase activities were significantly decreased in bromate-treated group, but change in renal glutathione reductase activity was not significantly different from any other experimental group.

Biological Activities of Acidic Polysaccharide of Korean Red Ginseng.111.-Effects on Metabolizing Activities in Acetaminophen- treated Rats (홍삼 산성다당체의 생리활성 연구(111)-아세트아미노펜 처리 흰쥐의 대사기능에 미치는 영향)

  • 이정규;최종원
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.267-273
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    • 1998
  • Pretreatment of acidic polysaccharide of Korean red ginseng (AcPS) for two weeks remarkably lowered the elevated content of lipid peroxide and levels of aminotransferases, sorbitol dehydrogenase, ${\gamma}$-glutamyltransferase, alkaline phosphatase and lactate dehydrogenase in liver intoxicated by acetaminophen (AA) . Pretreatments of AcPS also strengthen the liver function of glutathione related detoxication system indicated by glutathione contents and activities of glutathione S-transferase and glutathione reeducates which were affected by AA treatments. Activity of ${\gamma}$-glutamylcysteine syntheses was not changed by AcPS pretreatment whereas the activity of flu tathione reeducates was increased significantly. These results collectively indicate that the treatments of AcPS can promote the metabolism of lipid and reduce the production of peroxide in acetaminophen-intoxicated animals.

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Reduction of Hepatic Glutathione by Acute Taurine Treatment in Male Mice (숫컷 생쥐에서 타우린 투여에 의한 간내 글루타치온의 감소)

  • 이선영;곽혜은;김영철
    • YAKHAK HOEJI
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    • v.47 no.4
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    • pp.218-223
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    • 2003
  • Effect of taurine treatment on metabolism of glutathione (GSH) was studied in adult male ICR mice. An acute injection of taurine (250 mg/kg, ip) resulted in a significant decline of hepatic GSH level at t = 6 hr, but plasma GSH level was not altered. The activity of GSH-related enzyme in liver, such as GSH peroxidase, GSSG reductase, GSH S-transferases, ${\gamma}$-glutamylcysteine synthetase or ${\gamma}$-glutamyltranspeptidase, was not affected by taurine at t = 2.5 or 6 hr. Plasma cysteine and cystine levels were elevated rapidly following taurine treatment. Hepatic cysteine level was decreased by taurine, reaching a level approximately 70% of control at t = 4 and 6 hr. In conclusion, the results indicate that an acute dose of taurine decreases hepatic GSH level by reducing the availability of cysteine, an essential substrate for synthesis of this tripeptide in liver. It is also suggested that taurine may decrease the cysteine uptake by competing with this S-amino acid for a non-specific amino acid transporter.

Effects of Bromate on the Glutathione Synthesis in Various Organs of Rats (Bromate가 흰쥐의 장기 Glutathione 함량에 미치는 영향)

  • 김나영;강혜옥;이무강;최종원
    • Journal of Life Science
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    • v.13 no.5
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    • pp.626-633
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    • 2003
  • The effects of bromate administration on glutathione were studied in rats. The contents of glutathione in the liver and kidney were significantly decreased but the alteration was not significant in lung and blood by bromate adminstration. The decrease occurred without concomitant increases in oxidized glutathione (GSSG) or in the GSSG/GSH+GSSG ratio. The activities of $\gamma-glutamyl$ cysteine synthetase in the liver and kidney were decreased by bromate administration. $\gamma-Glutamyl$ transpeptidase activities was significantly decreased in the kidney and not significantly decreased in the lung of bromate treated-rats. These results suggest that the decreased synthesis of glutathione by bromate may be an important reason for the decreased level of glutathione in the liver and kidney, thus the decreased glutathione transport would be a factor on the changes of glutathione contents in bromate-treated rats.

Effect of Teminalia chebula Extract on Liver in Rat (가자(Terminalia chebula) 추출물이 흰쥐의 간장 활성에 미치는 영향)

  • 박종옥;이인섭;최종원
    • Journal of Life Science
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    • v.14 no.1
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    • pp.141-147
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    • 2004
  • In this study, we investigated the effect of Teminalia Chebula (TC) water extract on liver in Rat. Treatment of TC water extract was orally administered 200, 300 mg/kg daily for one week and two weeks. The clinical parameters of serum, values of AST, ALT showed significantly higher than in normal group. Xanthine oxidase and aldehyde oxidase activities were significantly increased comparison with normal group. Microsomal enzymes, aminopyrine N-demethylase and aniline hydroxylase were not affected. Water extract of TC also increased hepatic rnalondialdehyde formation and reduced glutathione content. We also found that water extract of TC decreased activities of glutathione S-transferase and glutathione reductase but was not affected activities of $\gamma$-glutamylcysteine synthetase Thus, it seems that the water extract of TC induced decrease of oxygen free radical scavenger, glutathione content by inhibition of glutathione reductase which may reform oxidized glutathione to reduced glutathione.

Gene Expression of Enzymes Related to Glutathione Metabolism in Anticancer Drug-resistant L1210 Sublines (항암제 내성 L1210세포의 Glutathione 대사 관련효소 유전자의 발현 양상)

  • Kim, Seong-Yong;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.32-47
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    • 1995
  • Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of ${\gamma}$-glutamylcysteine synthetase(GCS), ${\gamma}$-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR, L1210VcR, or L1210Cis) sublines were measured. Expression and amplification of GCS, GGT, and GST-${\pi}$ genes were also observed in the parent Ll210 and the drug-resistant Ll210 sublines. The concentration of GSH was increased 5.34 fold in L1210Cis, 2.83 fold in L1210VcR, and 1.78 fold in L1210AdR, compared to L1210. The activities of GCS and GGT were increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to Ll210. Expression of GCS, GGT, and GST-${\pi}$ genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-${\pi}$ were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

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CRYSTALLIZAT10N OF $\gamma$-GLUTAMYLCYSTEINE SYNTHETASE FROM Escherichia coli (대장균주로부터 분리한 GSH-1 효소의 결정화)

  • 황광언;김경규
    • Korean Journal of Crystallography
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    • v.4 no.2
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    • pp.100-104
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    • 1993
  • Reduced glutathione (GSH) plays a vital role in the metabolism of all cells. Glutathions, a tripeptide cowfosed of glutamic acid, cysteane, and gtycina is synthesized by two synthesized reutions. The first is catalyzed by Y-glutamylcysteine synthetase (GSH-I) and the second by glutathione synthetase (GSH-ll). The glutathione biosynthetic pathway of E. coziis mainly controlled by nonallosteric feedback inhibition of GHS-I by GSH. Determination of the three-dimensional structure of GSH-I by X-ray crystallography is necessary in order to understand the structure-function relationship at the molecular level. As the (irst step toward its structure determination, crystallization of 5. coli V-glutamylcystfine synthetase (GSH-I) has been achived using the hanging drop vapor diffusion method and capillaw method. Crystals of GSH-I have been grown from ammonium sulfate solution. The crystals grew at room temperature within 10 days to dimensions of 0.2 m x 0.2 m x 0.2 ml by hanging drop vapor diffusion method and diffracted to about 4 A resolution using synchrotron X-rays. Another crystal, grown by the capillary method to dimensions of 0.25 mm x 0.25 mm x 0.3 mm within 40 days, diffracted to about 4 A resolution using X-rays from a rotating anode.

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Biological Activity of Acidic Polysaccharide of Korean Red Ginseng 1.-Effects on Alcohol Detoxification System in the Llver of Alcohol-intoxicated Rats (홍삼 산성다당체의 생리활성 연구(I)-알코올 중독 동물의 간장 알코을 해독계에 미치는 영향)

  • 이정규;최종원
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.260-266
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    • 1998
  • The effects of acidic polysaccharide of Korean red ginseng (AcPS) on metabolisms of drug and alcohol in the liver were investigated. We could find that treatment of AcPS to six-week ethanol administered rats lowered the levels of alcohol and acetaldehyde in serum. We also we found that treatment of AcPS normalized the elevated activities of free radical generation system, decreased activities of detoxification system such as ${\gamma}$-glutamylcysteine synthetase and glutathione S-transferase, and decreased activities of acetaldehyde metabolizing system. The cytosolic alcohol dehy drogenase and microsomal ethanol oxidizing system (MEOS) were strongly enhanced.

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