• Title/Summary/Keyword: Copper,zinc-superoxide dismutase

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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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Molecular Characterization and Expression of CuZn-superoxide Dismutase (PSOD1) from Populus alba${\times}$Populus glandulosa

  • Lee Jun-Won;In Jun-Gyo;Lee Bum-Soo;Choi Yong-Eui;Kim Jin-Ju;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.1
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    • pp.52-59
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    • 2005
  • A cDNA, PSOD1, encoding cytosolic copper/zinc superoxide dismutase (CuZn-SOD) was cloned and characterized from a full length cDNA library prepared from Populus alba${\times}$Populus glandulosa cultured in vitro. A PSOD1, is 725 nucleotides long and has an open reading frame of 459 bp with 152 amino acid residues (pI 5.43). The deduced amino acid sequence of PSOD1 perfect matched to the previously reported CuZn-SOD (CAC33845.1). Consensus amino acid residues (His-45, -47, -62, -70, -79, -119) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of PSOD1 exhibited the high level of similarity from 100 to $85\%$ among previously registered SOD genes. The expression of PSOD1 in poplar increased at the 1 mM $H_{2}O_2$ and drought stress during 30 min and 60 min, but the ozone treated poplar increased at 30 min in the early time and then decreased at 60 min.

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Effect of Myricetin on mRNA Expression of Different Antioxidant Enzymes in B16F10 Murine Melanoma Cells (B16F10 Murine Melanoma Cell에서 Myricetin이 항산화효소의 m-RNA 발현에 미치는 영향)

  • Yu Ji Sun;Kim An Keun
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.86-91
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    • 2005
  • Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiinflammatory, antihistaminic, antioxidant and free-radica 1 scavenging abilities. The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress insults. To determine whether flavonoid, myricetin can exert antioxidative effects not only directly by modulating the AOE system but also scavenging free radical, we investigated the influence of the flavonoid myricetin on cell viability, different antioxidant enzyme activities, ROS level and the expression of different antioxidant emzyme in B16F10 murine melanoma cells. Myricetin in a concentration range from 6.25 to $50\;{\mu}M$ decreased superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities, but catalase (CAT) activity was increased. In the myricetin-treated group, ROS levels were decreased dose-dependently. Antioxidant enzyme expression was measured by RT-PCR. Myricetin treatment of B16F10 cells increased catalase expression. Expression levels of copper zinc superoxide dismutase (CuZn SOD) were not affected by exposure of myricetin. Manganese superoxide dismutase (Mn SOD) and GPx expression levels decreased slightly after myricetin treatment. In conclusion, the antioxidant capacity of myricetin was due to CAT and free-radical scavenging.

Purification and Characterization of Iron-Containing Superoxide Dismutase from Lentinus edodes

  • Park, Sang-Shin;Hwang, Soo-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.854-860
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    • 1999
  • Superoxide dismutase (SOD) was purified to homogeneity from fruiting bodies of edible mushroom, Lentinus edodes, by ammonium sulfate precipitation, diethylaminoethyl (DEAE)-Sepharose FF ion-exchange chromatography, Sephacryl S-200 gel filtration chromatography, and preparative PAGE. The molecular weight of the purified enzyme was estimated to be approximately 54 kDa by gel filtration chromatography, and the enzyme was shown to be consisted of two identical subunits of molecular weight 27 kDa by SDS-PAGE. The isoelectric point of the enzyme was 4.9 as determined by isoelectric focusing. The enzyme had optimal pH and temperature of pH 8.0 and $20^{\circ}C$, respectively. The activity of the enzyme was inhibited by hydrogen peroxide, but inhibited less by cyanide and azide. The native enzyme was found to contain 0.89g-atom of iron, 0.75g-atom of zinc, and 0.46g-atom of copper per mol of enzyme. Analysis of amino acids composition revealed that the SOD from L. edodes contained a relatively large amount of glutamic acid/glutamine, proline, cysteine, isoleucine, and leucine, but only a small amount of aspartic acid/asparagine, tyrosine, and tryptophan when compared to the other iron-containing SODs.

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Comparison of genetic structure of the Cu,Zn superoxide dismutase (SOD1) from Cordyceps militaris, Paecillomyces tenuipes and P.sinensis

  • Park, Nam-Sook;Lee, Sang-Mong;Sohn, Hung-Dae;Jin, Byung-Rae
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.71-74
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    • 2003
  • Superoxide dismutase (SOD), one of the essential element of the antioxidant defense system, mainly removes $O^{-10}$ $_2$ and also prevents $O^{-10}$ $_2$ mediated reduction of iron and subsequent OH$^{-10}$ generation, which is highly toxic to the organism. Of these SOD enzymes, Cu, Zn-containing SOD (SODI) is an important component of the antioxidant defense system in eucaryotic cells. The SODI enzyme binds one copper and one zinc ion and displays the Greek Key $\beta$-barrel fuld. (omitted)

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The protective effects of trace elements against side effects induced by ionizing radiation

  • Hosseinimehr, Seyed Jalal
    • Radiation Oncology Journal
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    • v.33 no.2
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    • pp.66-74
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    • 2015
  • Trace elements play crucial role in the maintenance of genome stability in the cells. Many endogenous defense enzymes are containing trace elements such as superoxide dismutase and metalloproteins. These enzymes are contributing in the detoxification of reactive oxidative species (ROS) induced by ionizing radiation in the cells. Zinc, copper, manganese, and selenium are main trace elements that have protective roles against radiation-induced DNA damages. Trace elements in the free salt forms have protective effect against cell toxicity induced by oxidative stress, metal-complex are more active in the attenuation of ROS particularly through superoxide dismutase mimetic activity. Manganese-complexes in protection of normal cell against radiation without any protective effect on cancer cells are more interesting compounds in this topic. The aim of this paper to review the role of trace elements in protection cells against genotoxicity and side effects induced by ionizing radiation.

Over-expression of Cu/ZnSOD Increases Cadmium Tolerance in Arabidopsis thaliana

  • Cho, Un-Haing
    • Journal of Ecology and Environment
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    • v.30 no.3
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    • pp.257-264
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    • 2007
  • Over-expression of a copper/zinc superoxide dismutase (Cu/ZnSOD) resulted in substantially increased tolerance to cadmium exposure in Arabidopsis thaliana. Lower lipid peroxidation and $H_2O_2$ accumulation and the higher activities of $H_2O_2$ scavenging enzymes, including catalase (CAT) and ascorbate peroxidase (APX) in transformants (CuZnSOD-tr) compared to untransformed controls (wt) indicated that oxidative stress was the key factor in cadmium tolerance. Although progressive reductions in the dark-adapted photochemical efficiency (Fv/Fm) and quantum efficiency yield were observed with increasing cadmium levels, the chlorophyll fluorescence parameters were less marked in CuZnSOD-tr than in wi. These observations indicate that oxidative stress in the photosynthetic apparatus is a principal cause of Cd-induced phytotoxicity, and that Cu/ZnSOD plays a critical role in protection against Cd-induced oxidative stress.

Characterization of Copper/Zinc-Superoxide Dismutase (Cu/Zn-SOD) Gene from an Endangered Freshwater Fish Species Hemibarbus mylodon (Teleostei; Cypriniformes)

  • Lee, Sang-Yoon;Kim, Keun-Yong;Bang, In-Chul;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.43-54
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    • 2011
  • Gene structure of copper/zinc-superoxide dismutase (Cu/Zn-SOD; sod1) was characterized in Hemibarbus mylodon (Teleostei; Cypriniformes), an endangered freshwater fish species in Korean peninsula. Full-length cDNA of H. mylodon SOD1 consisted of a 796-bp open reading frame sequence encoding 154 amino acids, and the deduced polypeptide sequence shared high sequence homology with other orthologs, particularly with regard to metal-coordinating ligands. Genomic structure of the H. mylodon sod1 gene (hmsod1; 1,911 bp from the ATG start codon to the stop codon) was typical quinquepartite (i.e., five exons interrupted by four introns); the lengths of the exons were similar among species belonging to various taxonomic positions. The molecular phylogeny inferred from sod1 genes in the teleost lineage was in accordance with the conventional taxonomic assumptions. 5'-flanking upstream region of hmsod1, obtained using the genome walking method, contained typical TATA and CAAT boxes. It also showed various transcription factor binding motifs that may be potentially involved in stress/immune response (e.g., sites for activating proteins or nuclear factor kappa B) or metabolism of xenobiotic compounds (e.g., xenobiotic response element; XRE). The hmsod1 transcripts were ubiquitously detected among tissues, with the liver and spleen showing the highest and lowest expression, respectively. An experimental challenge with Edwardsiella tarda revealed significant upregulation of the hmsod1 in kidney (4.3-fold) and spleen (3.1-fold), based on a real-time RT-PCR assay. Information on the molecular characteristics of this key antioxidant enzyme gene could be a useful basis for a biomarker-based assay to understand cellular stresses in this endangered fish species.