• Title/Summary/Keyword: Cu,Zn SOD

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Effect of Copper Ion on Oxygen Damage in Superoxide Dismutase-Deficient Saccharomyces Cerevisiae

  • Lee, Jeong-Ki;Kim, Ji-Myon;Kim, Su-Won;Nam, Doo-Hyun;Yong, Chul-Soon;Huh, Keun
    • Archives of Pharmacal Research
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    • v.19 no.3
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    • pp.178-182
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    • 1996
  • Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxidative stresses induced by 0.1 mM of copper ion $(Cu^{++})$ was studied. In aerobic culture condition, yeasts lacking MnSOD (mitochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared withwild type were observed under anaerobic condition. It was found that, under aerobic condition, the supplementation of 0.1 mM copper ioh:(Cu") into culture medium caused the remarkable increase of CuZnSOD but not so significant change in MnSOD. It was also observed that catalase activities appeared to be relatively high in the presence of copper ion in spite of the remarkable reduction of glutathion peroxidase in CuZnSOD-deficient yeasts, but the slight increments of catalase and glutathion peroxidase were detected in MnSOD-deficient strains. It implies that the lack of cytoplasmic SOD could be compensated mainly by catalase. However, these phenomena resulted in the significantincrease of cellular lipid peroxides content in CuZnSOD-deficient yeasts and the slight increment of lipid peroxides in MNSOD-deficient cells. In anaerobic cultivation supplementing copper ion, the cellular enzyme activities of catalase and glutathion peroxidase in SOD-deficient yeasts were slightly increased without any significant changes of lipid peroxides in cell membrane. It suggests that a little amount of free radicals generated by copper ion under anaerobic condition could be sufficiently overcome by catalase as well as glutathion peroxidase.dase.

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Photosynthetic Efficiency in Transgenic Tobacco Plants Expressing both CuZnSOD and APX in Chloroplasts against Oxidative Stress Caused by Highlight and Chilling (CuZnSOD와 APX를 엽록체에 발현시킨 담배식물체의 Highlight와 Chilling 스트레스에 대한 광합성 효율)

  • Kim, Yun-Hee;Kwon, Suk-Yoon;Bang, Jae-Wook;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.399-403
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    • 2003
  • In order to understand the protection effects of antioxidant enzymes against oxidative stress caused by various environmental stresses, transgenic tobacco (Nicotiana tabacum cv, Xanthi) plants expressing both copper/zinc superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) in chloroplasts (referred to as CA plants) were subjected to highlight (1,100$\mu$mol m$^{-2}$ sec$^{-1}$) and chilling at 4$^{\circ}C$. The protection effects of CA plants using leaf discs were compared with those of transgenic plants expressing either CuZnSOD or APX in chloroplasts (SOD plants or APX plants, respectively) and non-transgenic (NT) plants. CA plants showed about 15% protection in the photosynthetic efficiency (Fv/Fm) of photosystem II relative to NT plants 1 hr after treatment of both highlight and chilling, whereas they showed about 23% protection in the redox state of P700 in photosystem I at 3 hr after treatment. SOD plants or APX plants showed an intermediate protection effect between CA plants and NT plants. These results demonstrated that the coexpression of CuZnSOD and APX in chloroplasts importantly involves in the protection effects against oxidative stress caused by various environmental stresses.

Role of Inducibility of Superoxide Dismutases and Metallothionein of Mouse Lungs by Paraquat in Aging (Paraquat에 의한 생쥐 폐의 Superoxide Dismutases와 Metallothionein의 유도능과 노화와의 관계)

  • Lee, Tae-Bum;Park, Yoo-Hwan;Choi, Cheol-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.579-590
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    • 2001
  • Background : The aging process may be induced, at least in part, by reactive oxygen species(ROS). It has been thought that the lung could be a good source of ROS because it has a high oxygen tension. In the present study, we invetigated the inducibility of the first and last lines against oxidative stress, superoxide dismutases(CujZn-SOD and Mn-SOD) as a scavenger of ${O_2}^-\;{\cdot}$ and metallothionein(MT) as a scavenger of $OH{\cdot}$, respectively, in mouse lungs with age. Methods : Oxidative stress was induced by paraquat, an intracellular superoxide generator, at 1, 4, 8, and 12 months of age and then SODs and MT mRNAs were determined by RT-PCR method. Results : The steady-state level of Mn-SOD mRNA increased from 1 to 8 months but decreased thereafter. However, Mn-SOD mRNA was not induced by paraquat after 1 month. On the other hand, there was no change in the steady-state level of Cu/Zn-SOD mRNA, which decreased abruptly at 12 months of age. Additionally, Cu/Zn-SOD mRNA was not induced by paraquat at any age. There was no change in the steady-state level of MT mRNA with age whereas its inducibility by paraquat was intact at all ages. Conclusion : These results indicate that lack of induction of SODs with age may be one of the causative factors in the aging process while induction of MT may play an important role in the defense against oxidative stress. It is therefore implicated that the tissue antioxidant/prooxidant balance could be one of determinants of mean life span.

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Effects of Fermented Mulberry Leaves (Morus alba L.) on Oxidative Modification of Antioxidnat Enzymes (항산화 효소의 산화적 변형에 뽕잎 발효물이 미치는 영향)

  • Kang, Jung Hoon
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.3
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    • pp.985-994
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    • 2019
  • Muberry (Morus alba L.) leaves fermented with Hericium erinaceum mycelium (MA-HE) were assessed for the protection against oxidative modification of antioxidant enzymes, Cu,Zn-superoxide dismutase(SOD) and ceruloplasmin(CP). MA-HE were shown to significantly inhibited oxidative modifications and inactivations of Cu,Zn-SOD and CP induced by peroxyl radical. Antioxidant activity of MA-HE evaluated using peroxyl radical scavenging assays. MA-HE showed 44.03% of peroxyl radical scavenging activity at $100{\mu}g/mL$. Thus, MA-HE protect the antioxidant enzymes from oxidative damage by the scavenging peroxyl radicals. The results suggested that MA-HE was effectively removed reactive oxygen species in cells, thereby protecting cytotoxicity caused by oxidative stress.

High Glucose Potentiates the Alloxan-induced Cytotoxicity in Cultured Rat Insulinoma Cells (흰쥐 인슐린종세포에서 고농도 포도당의 Alloxan 독성 증강 효과)

  • 이병래;차종희;박재윤;고춘남;박평심
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.875-880
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    • 2000
  • Reactive oxygen species are produced under diabetic conditions and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study was to examine the effects of high glucose on the alloxan-induced beta cell injury. The insulinoma (RINm5F) cells were clutured either with high glucose (22.2 mM) or normoglucose (5.6 mM) in RPMI 1460 media for 3 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The effects of high glucose on the cytotoxicity of alloxan were also investigated in RINm5F cells and the cells viability were determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methods. Results showed that the CuZn-SOD activity was decreased but Mn-SOD activity was increased significantly in RINm5F cells cultured with high glucose (22.2 mM) media. The cytotoxicity of alloxan was increased by high glucose compared with normoglucose in RINm5F cells. Diethyl-dithiocarbarmate (DDC), as inhibitor of CuZn-SOC, also potentiate the alloxan-induced cytotoxocity in RINm5F cells. These results suggest that, in RINm5F cells, short term culture with high glucose media decreases Cu-Zn-SOD activity and the decreased activity of CuZn-SOD many one of the causative factors of beta-cell injury induced by high glucose.

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Hydroxyl Radical-Generating Function of Horseradish Cu,Zn-Superoxide Dismutase

  • Eum, Won-Sik;Kwon, Oh-Bin;Kang, Jung Hoon
    • BMB Reports
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    • v.31 no.5
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    • pp.492-497
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    • 1998
  • Cu,Zn-superoxide dismutase (SOD) was purified from horseradish by using Mono Q and Superose 12 FPLC column chromatography. The native molecular mass of the purified enzyme was approximately 33 kDa, as determined by gel filtration. The subunit molecular weight, as estimated by SDS-PAGE, was 16 kDa. These results indicated that the native enzyme is a homodimer. We investigated the free radical-generating function of horseradish Cu,Zn-SOD by using a chromogen, 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate) (ABTS) which reacts with ${\cdot}OH$ radicals to form $ABTS^{+{\cdot}}$ The formation of $ABTS^{+{\cdot}}$ was required for both active Cu, Zn-SOD and $H_2O_2$. The optimal pH for the free radical-generating activity of this enzyme was 6.0-8.0, and it retained about $40^{\circ}C$ of its maximum activity when exposed at $40^{\circ}C$ for 15 min. A neutral scavenger, ethanol, inhibited the $ABTS^{+{\cdot}}$ formation by horseradish Cu, Zn-SOD more effectively than that by the mammalian enzyme. These results suggest that the active channel of horseradish enzyme is slightly larger than that of the mammalian enzyme.

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Effect of Metals on Anti-Oxidase Activity and Isozyme patterns in Brassica juncea

  • Jeong, Hyung-Jin;Lee, In-Jung;Sung, Mi-Hyang
    • Korean Journal of Plant Resources
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    • v.10 no.3
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    • pp.235-240
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    • 1997
  • To study the effects of metal ions on the activity of anti-oxidase enzymes, the activity of superoxide dismutase (SOD) and peroxidase (POD) and isozyme patterns of Brassica juncea have been studied after treating with CD, Cu, Zn, and Al. The activity of SOD after treating with metal ions was higher than that of untreated control. SOD activity in leaves increased by treatment of 50 ppm of Zn and 500 ppm of Al. POD in stems gave highest activity after treating with 500 ppm of Cu. When the activity was compared by plant parts, lowest POD activity was observed in leaves in which protein content was higher than other tissues. When the activity was expressed as percentage of control, SOD activity was increased after treating with metal ions. SOD activity in leaves and roots of metal treated plant was significantly increased under the metal ions stress conditions. In the roots of 50 ppm of Zn treated plant, SOD activity was extremly high. POD activity was inhibited with Cd and Zn treatment in all parts of the plant. However, in leaves and stems, there was marked increase in activity after treating with Cu. The patterns of SOD isozyme after metal treatment show that two bands were stained in all metal ion treated and that no new band appeared. POD isozyme band intensity resulting from the treatment of metal ions was in order of roots > stems > leaves, but there was no significant difference.

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Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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Effect of Metals on Anti - Oxidase Activity in Persicaria vulgaris Webb. et Moq. (중금속이 봄여뀌(Persicaria vulgaris Webb. et Moq.)의 항산화효소활성에 미치는 영향)

  • Sung, Mi-Hyang;Jeong, Hyung-Jin;Kim, Kun-Woo;Kwak, Sang-Soo
    • Korean Journal of Weed Science
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    • v.16 no.4
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    • pp.346-353
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    • 1996
  • To study the effects of metal ions on the activities of antioxidative enzymes, the activities of superoxide dismutase(SOD), peroxidase(POD), catalase(CAT) of Persicaria vulgaris has been studied after treating with Cd, Cu, Zn and Al. 1. The activities of SOD in leaf and stem were decreased, but that in root was increased. Among the metal ions studied in this report, Al gave the highest increase in SOD activity in root. 2. The activities of POD after treating with Cd or Cu did not show any significant differences. POD activities after treating with Zn and Al has been decreased, however, that in root showed increased activities after treating with Zn 5,000 ppm or Al 500 ppm. 3. The activity of CAT in leaf was decreased with every metals studied. The CAT activity in root was increased with increased concentration. The root treated with Al showed highest activity. 4. The presence of isozymes after treated metal ions has been studied in gel electrophoresis. The POD treated plant did not show any new isozymes, but the intensity of one of pre-existent band was increased. The SOD treated plant showed the several new isozymes.

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Oxygen Toxicity of Superoxide Dismutase-Deficient Saccharomyces cerevisiae by Paraquat (Paraquat에 의해 유도된 Superoxide Dismutase 결핍 효모의 산소 독성)

  • 김지면;남두현용철순허근
    • KSBB Journal
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    • v.10 no.5
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    • pp.561-567
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    • 1995
  • Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

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