• Title/Summary/Keyword: $sod2^{+}$

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Molecular Cloning and Expression of Sequence Variants of Manganese Superoxide Dismutase Genes from Wheat

  • Baek, Kwang-Hyun;Skinner, Daniel Z.
    • Korean Journal of Environmental Agriculture
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    • v.29 no.1
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    • pp.77-85
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    • 2010
  • Reactive oxygen species (ROS) are very harmful to living organisms due to the potential oxidation of membrane lipids, DNA, proteins, and carbohydrates. transformed E.coli strain QC 871, superoxide dismutase (SOD) double-mutant, with three sequence variant MnSOD1, MnSOD2, and MnSOD3 manganese superoxide dismutase (MnSOD) gene isolated from wheat. Although all QC 871 transformants grown at $37^{\circ}C$ expressed mRNA of MnSOD variants, only MnSOD2 transformant had functional SOD activity. MnSOD3 expressed active protein when grown at $22^{\circ}C$, however, MnSOD1 did not express functional protein at any growing and induction conditions. The sequence comparison of the wheat MnSOD variants revealed that the only amino acid difference between the sequence MnSOD2 and sequences MnSOD1 and 3 is phenylalanine/serine at position 58 amino acid. We made MnSOD2S58F gene, which was made by altering the phenylalaine to serine at position 58 in MnSOD2. The expressed MnSOD2S58F protein had functional SOD activity, even at higher levels than the original MnSOD2 at all observed temperatures. These data suggest that amino acid variation can result in highly active forms of MnSOD and the MnSOD2S58F gene can be an ideal target used for transforming crops to increase tolerance to environmental stresses.

Identification and Characterization of SOD Isoenzymes in Acanthopanax koreanum Plants (섬오갈피나무에서 SOD Isoenzyme의 식별 및 특성규명)

  • 오순자;박영철;김응식;고석찬
    • Korean Journal of Plant Resources
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    • v.12 no.3
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    • pp.234-239
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    • 1999
  • The isoenzyme patterns and activities of superoxide dismutase(SOD) were investigated from leaves of Araliaceae plants. Of the eight isoenzymes, two isoenzymes(SOD 4 and SOD 6) were prevalent to leaves of Araliaceae plants. The patterns of these two isoenzymes were most various in the leaves of Acanthopanax senticosus for. inermis, while their activity was highest in the leaves of A. koreanum. These two isoenzymes were respectively identified as Fe-SOD and CuZn-SOD, based on selective inhibition with KCN or$H_2O_2$. The SOD isoenzyme patterns did not differed among stem barks, root barks and leaves of A. koreanum. However, the activities of Fe-SOD and CuZn-SOD were higher in the root bark and in leaves, respectively. Both of Fe-SOD and CuZn-SOD were stable for 1 hr at 30-4$0^{\circ}C$, while unstable above 5$0^{\circ}C$.

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Expression of Human SOD1 and Mutant SOD1 (G93A) in E. coli and Identification of SOD1 as a Substrate of HtrA2 Serine Protease (대장균에서의 human SOD1과 mutant SOD1 (G93A) 단백질의 발현과 HtrA2의 기질 여부 확인에 관한 연구)

  • Kim, Goo-Young;Kim, Sang-Soo;Park, Hyo-Jin;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.5
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    • pp.716-722
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    • 2006
  • Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.

The Study of Superoxide dismutase (SOD) and SOD-mimic Compounds in Panax ginseng C.A.Meyer

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    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.188-193
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    • 1997
  • Panax ginseng C.A.Meyer,1 to 5 years old were electrophored and were stained for SOD activity. The result indicated a total of 13 distinct form of the enzyme and the pattern of achromatic bands were not different according to ages. Nine of the enzyme activities were eliminated with cyanide or peroxide treatment and were resistant to treatment of chloroform plus ethanol. It suggested that they may be cupro-zinc containing SOD, whereas four were cyanide or peroxide resistant and were eliminated with cholroform plus ethanol treatment. They may be manganese containing SOD. Ginseng roos. 1 to 5 years old were analyzed for their SOD measurement of SOD activities of all extracts, the significant difference of SOD activities were not shown according to ages. All ginseng extracts had the total SOD activities of all extracts, the significant difference of SOD activities were not shown according to ages. All ginseng extracts had the total SOD activities of about 700-800 unit/g of fresh weight. Therefore, the SOD activities from SOD-mimic compounds were higher than one from SOD. The ratio between the SOD activity from SOD-mimic compounds and one from true SOD was approximately 2:1 to 3:1.

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Functional Characteristics of Cytoplasmic and Periplasmic Photobacterium leiognathi CuZnSOD (PSOD) in Escherichia coli SOD Double Mutants (Escherichia coli SOD 이중 돌연변이체내에서 세포질과 Periplasm에 분포하는 Photobacterium leiognathi CuZnSOD(PSOD)의 기능적 특성)

  • Kim, Young-Gon;Yang, Mi-Kyung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.139-146
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    • 1994
  • Protective effects on subcellular localization of Photobacterium leiognathi CuZnSOD(PSOD) were examined in Escherichia coli SOD mutant cells on the treatment of paraquat, heat shock $(37^{\circ}C{\to}42^{\circ}C{\to})$, hydrogen peroxide and copper sulfatem respectively. The physiological characteristics of the periplasmic and cytoplasmic PSOD localized differently are dependent on the conditions in this experiment. Cells expressing SOD periplasmically in the treatments of paraquat and $H_2O_2$ respectively were somewhat better protective effects cells expressiong SOD cytoplasmically at comparable level and SOD expression level showed, the most consistently important variable. However, this was reversed in the treatments of heat shock and $CuSO_4$, respectively.

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Induction of antioxygenic enzymes as defense systems in plant cells against low temperature stress : (II) $Mn^{+2}-induced$ SOD activation and enhancement of cold tolerance in rice seedlings (식물의 냉해에 대한 생체방어기구로서 항산소성 효소의 유도 : (II) $Mn^{+2}$이온에 의한 세포내 SOD의 활성화와 벼 유묘의 내냉성 향상)

  • Hahn, Chang-Kyun;Kim, Jong-Pyung;Jung, Jin
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.168-173
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    • 1991
  • The uptake of $Mn^{+2}$, a metal cofactor Mn-SOD, by rice seedings resulted in not only a substantial increase in SOD activity in leaf tissues of the plants, but also a significant enhancement of their cold tolerance : the relative extent of the cold tolerance appeared to accord with relative level of the SOD activity. In contrast, $Fe^{+3},\;Cu^{+2}$ and $Zn^{+2}$, which are the cofactors of Fe-SOD and Cu/Zn-SOD, were found to be ineffective for increasing the SOD activity as well as for improving the chilling-resistant capacity of the plants. The results suggest that Mn-SOD, which is most likely induced by its substrate(superoxide) and activated by the presence of $Mn^{+2}$a at high level, is the enzyme acting as an active component of the defense system against low temperature stress in rice plants. In addition, the application of abscisic acid which has been know to protect to some extent certain plants from chilling injury brought about an increase in SOD activity in rice tissues, providing another affirmative information for the crucial role of SOD under the circumstance of cold stress in plants.

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Comparison of Carbonaceous Sediment Oxygen Demand in Lake Paldang and Lake Chungju (팔당호와 충주호 퇴적물의 탄소성 산소요구량 비교)

  • Shin, Yu-Na;Park, Hae-Kyung;Lee, Sang-Won;Kong, Dong-Soo
    • Korean Journal of Ecology and Environment
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    • v.40 no.3
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    • pp.439-448
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    • 2007
  • The purpose of this study was to investigate the seasonal variations of sediment oxygen demand (SOD) in Lake Paldang and Lake Chungju of the Han River system and to suggest SOD values as parameters for the water quality prediction models of two lakes. SOD was measured at laboratory using sediment collected at 2 sites in Lake Paldang from June to November and at 4 sites in Lake Chungju from May to November in 2005, respectively. It was found from the laboratory test that the SOD in Lake Paldang ranged from 337.8 to 881.0 mg $O_2m^{-2}d^{-1}$ and in Lake Chungju ranged from 143.0 to 969.1 mg $O_2m^{-2}d^{-1}$. The SOD of Lake Paldang showed similar variations to the content of organic matter of sediment. The SOD of Lake Chungju was positively correlated with temperature (r=0.78, p<0.01), $PO_4-P(r=0.79,\;p<0.01)$, TP (r=0.55, p<0.05), DTP (r=0.55, p<0.05), $NO_3-N$ (r=(0.72, p<0.01) of hypolimnetic water. These results indicate that the SOD of Lake Paldang was affected by the content and origin of organic matter of sediment and the SOD of Lake Chungju was closely correlated with physical and chemical factors.

Characterization of Superoxide Dismutase in Lactococcus lactis

  • Chang, Woo-Suk;So, Jae-Seong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.732-736
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    • 1999
  • The superoxide dismutase (SOD) in Lactococcus lactis was measured quantitatively and qualitatively under various culture conditions. The L. lactis SOD was induced by oxidative stress. As the concentration of paraquat to produce superoxide radicals increased, the growth of L. lactis decreased with concomitant increase of SOD activity. The SOD activity was found to be growth-phase dependent: when aerobically grown cells entered to the stationary phase, the activity increased gradually until the late stationary phase. From inhibition studies, L. lactis SOD was found to be insensitive to KCN and $H_2O_2$ which are known to inhibit Cu/ZnSOD and FeSOD, respectively. Moreover, as the concentration of manganese in the medium increased, the activity of SOD also increased. These data strongly suggested that L. lactis possessed a single manganese-containing SOD (MnSOD). Finally, a putative sod gene fragment of 510 bp was identified in L. lactis using a polymerase chain reaction (PCR) with degenerate primers designed from the deduced DNA sequences of known SOD genes.

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The Effect of the SOD2 and SOD3 in Candida albicans on the Antioxidant System and its Potential as a Natural Antioxidant

  • Yeonju HONG;Min-Kyu KWAK
    • The Korean Journal of Food & Health Convergence
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    • v.10 no.2
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    • pp.13-17
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    • 2024
  • Oxygen is necessary to sustain life, but reactive oxygen species (ROS) produced by oxygen metabolism can cause mutations and toxicity. ROS can damage cellular macromolecules, leading to oxidative stress, which can accelerate cell death and aging. ROS generated in food affect the taste, color, and aroma of food, and high levels of ROS in meat can cause spoilage. Superoxide dismutase (SOD) plays an important role in scavenging ROS in food and reducing their toxicity to organisms. SOD exerts its antioxidant effect by catalyzing the breakdown of O2-• to H2O2. As a natural antioxidant, SOD has the ability to regenerate and maintain its activity over a long period of time without depletion, unlike chemical antioxidants that may have side effects or stability issues. This antioxidant effect of SOD has great potential in a variety of industries, and in the food industry it can be utilized to improve product quality and provide safe and healthy products to consumers. By disrupting the SOD2 and SOD3 genes in Candida albicans, we studied the effects of SOD2 and SOD3 genes on the antioxidant system, suggesting its potential as a natural antioxidant.

Zinc(II) ion promotes anti-inflammatory effects of rhSOD3 by increasing cellular association

  • Kim, Younghwa;Jeon, Yoon-Jae;Ryu, Kang;Kim, Tae-Yoon
    • BMB Reports
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    • v.50 no.2
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    • pp.85-90
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    • 2017
  • Recently, we demonstrated that superoxide dismutase 3 (SOD3) is a strong candidate for biomedicine. Anti-oxidant function of SOD3 was accomplished without cell penetration, and it inhibited the inflammatory responses via non-enzymatic functions. SOD3 has the heparin binding domain associating cell surface. Interestingly, we found that $Zn^{2+}$ promotes transduction effects of recombinant human SOD3 (rhSOD3) by increasing uptake via the heparin binding domain (HBD). We demonstrated an uptake of rhSOD3 from media to cell lysate via HBD, resulting in an accumulation of rhSOD3 in the nucleus, which was promoted by the presence of $Zn^{2+}$. This resulted in increased inhibitory effects of rhSOD3 on NF-{\kappa}B and STAT3 signals in the presence of $Zn^{2+}$, which shows elevated association of rhSOD3 into the cells. These results suggest that an optimized procedure can help to enhance the inflammatory efficacy of rhSOD3, as a novel biomedicine.