• Title, Summary, Keyword: CuZn-SOD

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Glycation of Copper, Zinc-Superoxide Dismutase and its Effect on the Thiol-Metal Catalyzed Oxidation Mediated DNA Damage

  • Park, Jeen-Woo;Lee, Soo-Min
    • BMB Reports
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    • v.28 no.3
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    • pp.249-253
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    • 1995
  • The nonenzymatic glycation of copper, zinc-superoxide dismutase (Cu,Zn-SOD) led to inactivation and fragmentation of the enzyme. The glycated Cu,zn-SOD was isolated by boronate affinity chromatography. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA and the generation of strand breaks in pBhiescript plasmid DNA by a metal-catalyzed oxidation (MCO) system composed of $Fe^{3+}$, $O_2$, and glutathione (GSH) as an electron donor was enhanced more effectively by the glycated CU,Zn-SOD than by the nonglycated enzyme. The capacity of glycated Cu,Zn-SOD to enhance damage to DNA was inhibited by diethylenetriaminepentaacetic acid (DETAPAC), azide, mannitol, and catalase. These results indicated that incubation of glycated CU,Zn-SOD with GSH-MCO may result in a release of $Cu^{2+}$ from the enzyme. The released $Cu^{2+}$ then likely participated in a Fenton-type reaction to produce hydroxyl radicals, which may cause the enhancement of DNA damage.

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Superoxide Dismutase Gene Expression Induced by Lipopolysaccharide in Alveolar Macrophage of Rat (폐포대식세포에서 내독소 자극에 의한 Superoxide Dismutase 유전자발현의 조절 기전)

  • Park, Kye-Young;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Hyun, In-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.522-534
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    • 1995
  • Background: In the pathogenesis of acute lung injury induced by lipopolysaccharide(LPS), oxygen radiclls are known to be involved in one part. Superoxide dismutase(SOD) protects oxygen radical-induced tissue damage by dismutating superoxide to hydrogen peroxide. In eukaryotic cells, two forms of SOD exist intracellularly as a cytosolic, dimeric copper/zinc-containing SOD(CuZnSOD) and a mitochondrial, tetrameric manganese-containing SOD(MnSOD). But there has been little information about SOD gene expression and its regulation in pulmonary alveolar macrophages(PAMs). The objective of this study is to evaluate the SOD gene expression induced by LPS and its regulation in PAMs of rat. Method: In Sprague-Dawley rats, PAMs obtained by broncholaveolar lavage were purified by adherence to plastic plate. To study the effect of LPS on the SOD gene expression of PAMs, they were stimulated with different doses of LPS($0.01{\mu}g/ml{\sim}10{\mu}g/ml$) and for different intervals(0, 2, 4, 8, 24hrs). Also for evaluating the level of SOD gene regulation actinomycin D(AD) or cycloheximide(CHX) were added respectively. To assess whether LPS altered SOD mRNA stability, the rate of mRNA decay was determined in control group and LPS-treated group. Total cellular RNA extraction by guanidinium thiocyanate/phenolfchlorofonn method and Northern blot analysis by using a $^{32}P$-labelled rat MnSOD and CuZnSOD cDNAs were performed. Results: The expression of mRNA in MnSOD increased dose-dependently, but not in CuZnSOD. MnSOD mRNA expression peaked at 8 hours after LPS treatment. Upregulation of MnSOD mRNA expression induced by LPS was suppressed by adding AD or CHX respectively. MnSOD mRNA stability was not altered by LPS. Conclusion: These findings show that PAMs of rat could be an important source of SOD in response to LPS, and suggest that their MnSOD mRNA expression may be regulated transcriptionally and require de novo protein synthesis without affecting mRNA stability.

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Hepatic Expression of Cu/Zn-Superoxide Dismutase Transcripts in Response to Acute Metal Exposure and Heat Stress in Hemibarbus mylodon (Teleostei: Cypriniformes)

  • Cho, Young-Sun;Bang, In-Chul;Lee, Il-Ro;Nam, Yoon-Kwon
    • Fisheries and aquatic sciences
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    • v.12 no.3
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    • pp.179-184
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    • 2009
  • Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at $5{\mu}M$ for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to $5{\mu}M$). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure ($5{\mu}M$) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to $30^{\circ}C$ at a rate of $1^{\circ}C/h$ followed by $30^{\circ}C$ for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.

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.

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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Screening of Korean Medicinal and Food Plants with Antioxidant Activity (한국 약용 및 식용식물들의 항산화성 식물탐색)

  • Chung, Il-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.311-322
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    • 1998
  • Sixty medicinal and food plants native to Korea were mainly selected with old traditional habit and antioxidant activity was investigated. The 80% EtOH extracts of sixty medicinal and food plants were screened for antioxidant activity. Antioxidant activity was measured by the TBA (Thiobarbituric acid), DPPH (1, 1-Diphenyl-2-picrylhydrazyl), SOD (superoxide dismutase) which was evaluated by the nitro blue tetrazolium(NBT) reduction method. Among sixty plants, black Glycine max(87. 3%) and Solanum nigrum (80.6%) exhibited the highest antioxidant activity by TBA and DPPH methods, respectively. Also, 10 species extracts including black Glycine max showed the high activity value in these two methods. The SOD characteristics on black Glycine max seed extracts which showed the highest SOD activity (53.5%) exhibited four major SODs; two Cu/ZnSODs and two FeSODs. However, Adenophaora vertidllata which showed lowest SOD value (10.4%) had only Cu/Zn SOD. No varietal differences in the high SOD value were detected in the Cu/Zn SOD isozyme patterns.

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Effects of Copper / Zinc-Containing Superoxide Dismutase (Cu, Zn-SOD) and Catalase on Paraquat-Induced Injury in Primary Cultured Rat Skin Fibroblast (일차 배양한 백서 피부섬유아세포에서 Paraquat 독성에 미치는 SOD 와 Catalase 의 영향)

  • Cha, Jong Hui;Yu, Ui Gyeong
    • Journal of the Korean Chemical Society
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    • v.38 no.1
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    • pp.74-79
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    • 1994
  • The participation of superoxide in initiating tissue damage derived from xenobiotics is best illustrated by paraquat intoxication. In the present study, the roles of superoxide dismutase and catalase on paraquat-induced cell injury were investigated using primary cultured rat skin fibroblast. The degree of cell injury was assassed by the conversion of reduced MTT to a blue formazan. Paraquat produced concentration-and time-related cell injury in cultured rat skin fibroblast. Paraquat induced-cell injury was aggravated by pretreatment of aminotriazol (AT: 10 mM), an catalase inhibitor, and attenuated by addition of catalase (100∼500 unit/ml). However, the effects of diethyldithiocarbamate (DDC : 10 mM), copper- and zinc-containing superoxide dismutase (Cu, Zn-SOD) inhibitor, and Cu, Zn-SOD on paraquat-induced injury were not significant. These results suggest that hydrogen peroxide might be more responsible factor than superoxide in the pathogenesis of paraquat-induced cell injury.

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Construction of Gene-Specific Primers for Various Antioxidant Isoenzyme Genes and Their Expressions in Rice (Oryza sativa L.) Seedlings Obtained from Gamma-irradiated Seeds

  • Kim, Jin-Hon;Chung, Byung-Yeoup;Kim, Jae-Sung;Wi, Seung-Gon;Yang, Dae-Hwa;Lee, Choon-Hwan;Lee, Myung-Chul
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.115-120
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    • 2004
  • For the expression study of antioxidant isoenzyme genes in rice (Oryza sativa L.) plants, extensive searches for genes of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) isoforms were performed through the GenBank database. The genes for two cytosolic and one plastidic CuZn-SOD, one Fe-SOD, two Mn-SOD, two cytosolic and two chloroplastic (stromal and thylakoid) APX, and three CAT isoforms were available in japonica-type rice. These isoforms were named as cCuZn-SOD1, cCuZn-SOD2, pCuZn-SOD, Fe-SOD, Mn-SOD1, Mn-SOD2, cAPXa, cAPXb, Chl_sAPX, Chl_tAPX, CATa, CATb, and CATc, respectively. Since they shared a high degree of homology in the nucleotide and amino acid sequences, the gene-specific primers for the genes were designed directly from their full-length cDNAs found in the database except for the CATa gene. These primers were used in the RT-PCR analysis to investigate the differential expression of antioxidant isoenzyme genes in rice plants from the seeds irradiated with low doses (2, 4, 8, and 16 Gy) of gamma-radiation. The gammairradiation slightly increased the transcripts of pCuZn-SOD, while those of Fe-SOD, cAPXb, and CATb decreased. However, no substantial differences were observed in the expression of all the isoenzyme genes between the control and irradiated groups. In this study, gene specific primers for thirteen SOD, APX and CAT isoenzymes were constructed from the full-length cDNAs. The results of RT-PCR analysis obtained by using these primers suggests that the expression levels of SOD, APX, and CAT isoenzyme genes in rice seedlings were hardly affected by gamma-irradiation at the seed stage.

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Genomic Structure of the Cu/Zn Superoxide Dismutase(SOD1) Gene from the Entomopathogenic Fungus, Cordyceps pruinosa

  • Park, Nam Sook;Jin, Byung Rae;Lee, Sang Mong
    • International Journal of Industrial Entomology
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    • v.39 no.2
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    • pp.67-73
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    • 2019
  • The genomic structure of the Cu/Zn superoxide dismutase (SOD1) gene from the entomopathogenic fungus, Cordyceps pruinosa was characterized. The SOD1 gene of C. pruinosa spans 947 nucleotides and consisted of four exons encoding for 154 amino acids and three introns. Four exons of the SOD1 gene are composed of 13, 331, 97 and 20 nucleotides respectively. Homology search of amino acid sequences of the SOD1 gene of C. pruinosa with another 13 fungi species showed higher sequence similarity of 69% ~ 95% and had the most highest sequence identity of 95% with Beauveria bassiana and Cordyceps militaris, which can easely infect domesticated Bombyx mori and another wild lepidopteran species in artificial or natual manner of infection. This SOD1 gene sequence showed copper, zinc and beta-barrel fold sites. Homology search showed that the Cu/Zn SOD1 gene from the entomopathogenic fungus, C. pruinosa is an orthologous gene homolog present in different species of organism whose ancestor predates the split between the relating species. In addition, C. pruinosa SOD1 gene is placed together within the ascomycetes group of fungal clade. From these results it is concluded that C. pruinosa SOD1 gene is orthologous gene having the same or very similar functions with a common evolutionary ancestor.

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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