• Title/Summary/Keyword: superoxide dismutase-1

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Isolation and Characterization of the sod2$^{2+}$ Gene Encoding a Putative Mitochondrial Manganese Superoxide Dismutase in Schizosaccharomyces bombe

  • Jeong, Jae-Hoon;Kwon, Eun-Soo;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.37-41
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    • 2001
  • The fission yeast Schizosaccharomyces pombe contains two distinct superoxide dismutase (SOD) activities, one in the cytosol encoded by the $sod2^{+}$ gene and the other in mitochondria. The $sod2^{+}$ gene encoding putative mitochondrial manganese superoxide dismutase (MnSOD) was isolated from the S. pombe genomic library using a PCR fragment as the probe. The nucleotide sequence of the $sod2^{+}$ gene and its flanking region (4051 bp HindIII fragment) was determined. An intron of 123 nt in size was predicted and confirmed by sequencing the cDNA following reverse transcription PCR. The predicted Sod2p consists of 218 amino acid residues with a molecular mass of 24,346 Da. The deduced amino acid sequence showed a high degree of homology with other MnSODs, especially in the metal binding residues at the active site and their relative positions. The transcriptional start site was mapped by primer extension at 231 at upstream from the ATG codon. A putative TATA box(TATAAAA) was located 58 nt upstream from the transcriptional start site and putative polyadenylation sites were located at 1000, 1062, and 1074 nt downstream from the ATG start codon.

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Effect of Geraniol on the Proliferation of L1210 Cells and ICR Mouse Macrophages, and the Activities of Superoxide Dismutase (SOD) and Inducible Nitric Oxide Synthase ( iNOS) Activities (Geraniol이 L1210 세포와 ICR 생쥐 대식세포의 증식,Superoxide Dismutase(SOD)와 Inducible Nitric Oxide Synthase(iNOS) 효소활성에 미치는 영향)

  • Kim, Ji-Yeon;Park, Sie-Won
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.309-316
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    • 2004
  • The present investigation was undertaken to find out the anticancer activity of monoterpene compounds. Monoterpenes showed generally the inhibitory effect on the proliferation o f L1210 cancer cells (cytotoxicity). Geraniol was found to exibit the most potent cytotoxic effect on L1210 cells with an IC50 values of $0.67{\mu}g/ml$. On the other hand, geraniol proved to be capable of stimulating the macrophage proliferation (135% of control). When the life prolonging activity of geraniol by daily oral administration of 0.1~10${\mu}g/10{\mu}l/20$ g body weight to Sarcoma 180 bearing ICR mouse was examined, there was also a significant elevation of survival (best result of 134% of control). The contradictory effects of geraniol on the proliferation of L1210 cells and macrophages proved to be accompanied by the coincident alterations of RNS (reactive nitrogen species) related enzymes activities such as inducible nitric oxide synthase (Inos) in macrophages and ROS (reactive oxygen species) related enzymes activities such as superoxide dismutase (SOD) in L1210 cells, respectively.

Effects of Dietary Cheongkukjang on Liver Superoxide Dismutase Activity of Parrotfish Oplegnathus fasciatus (사료 내 분말 청국장이 돌돔, parrootfish, Oplegnathus fasciatus의 간 내 superoxide dismutase 활성에 미치는 영향)

  • Pham, Minh Anh;Lee, Kyeong-Jun
    • Journal of Aquaculture
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    • v.20 no.2
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    • pp.132-139
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    • 2007
  • A four-week feeding trial was conducted to investigate the effects of dietary soybean meal (SBM) and powdered Cheongkukjang (CKJ) on non-specific immune responses of parrotfish Oplegnathus fasciatus. Three isonitrogenous (42% crude protein) and isocaloric (17.1 MJ/kg) diets were formulated to replace fish meal by 0, 25% SBM or 25% CKJ (designated as FM, 25SBM and 25CKJ, respectively). Ninety fish (initial body weight 122 g) were randomly allotted into nine 150 L tanks. One of the three experimental diets was fed to triplicate groups of fish for 4 weeks. After the feeding trial, no differences were observed in growth performances and feed utilization among fish groups. Liver superoxide dismutase activity of the fish fed CKJ containing diet was significantly higher than that of the control groups. DPPH radical scavenging and $Fe^{2+}-chelating$ activities of the experimental diets containing SBM or powdered CKJ were significantly higher than that of the control diet. The results of the present study suggest that dietary inclusion of powdered 25CKJ significantly increased liver superoxide dismutase activity and did not affect the growth performances, feed utilization, morphological parameters, as well as hematological values of parrotfish.

Antioxidant Property of the Gagam-Hyungbang-Gihwang-tang Using Biochemical Markers of Carcinogenesis (가감형방지황탕 열수 추출물이 항산화 작용에 미치는 영향)

  • Han Jin-Soo;Park Seong-Sik
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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    • pp.204-214
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    • 2005
  • Objectives : The verity extract of the Gagam-Hyungbang-Gihwang-tang (GHG) was assessed to determine the mechanisms of its antioxidant activity. Methods : The fellowing effects were measured : GHG exhibited a concentration-treatment; scavenging ${\alpha},\;{\alpha}-diphenyl-\beta-picrylhydrazyl$ (DPPH) radical, linoleic acid oxidation in a thiocyanate assay system, and superoxide anion, hydroxyl radical-induced DNA nicking. We investigated mRNA levels such as superoxide-dismutase. Results : The GHG extract showed dose-dependent free radical scavenging activity, including DPPH radicals, hydroxyl radicals, and superoxide anion, using different systems. The GHG was also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in Fenton's reaction mixture. Furthermore, SOD-1 mRNA expression levels increased in tat hepatoma H4IIE cells Conclusions : We expect that GHG will to helpful to the development of antioxidant activity treatments.

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Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli (사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구)

  • 이우길;김영호;양중익;노현모
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.91-97
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    • 1990
  • Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

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Changes of superoxide dismutase and glutathione peroxidase in light damaged rat retina

  • Kaidzu, Sachiko;Tanito, Masaki;Takanashi, Taiji;Ohira, Akihiro
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.430-432
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    • 2002
  • The changes in expression of copper-zinc superoxide dismutase (CuZn-SOD), manganese superoxide dismutase (Mn-SOD) and glutathione peroxidase (GPX) in light-damaged rat retinas were examined. Sprague-Dawley rats (male, 6-weeks-old) were maintained on a cyclic photoperiod (12 hours light and 12 hours darkness) for 2 weeks. The illumination intensity during the light period was 80 lux. To induce light damage to the retina, a high-intensity illumination (3000-lux) was applied to the animals for 24 hours. After light exposure, the animals were returned to cyclic lighting. Eyes were enucleated 12 and 24 hours after light exposure started or 1,3, and 7 days after light exposure ended. Eyes were fixed and embedded in paraffin wax. Tissues were cut into 4${\mu}{\textrm}{m}$-thick sections. Sections were immunostained using antibody against CuZn-SOD, Mn-SOD, GPX and 8-hydroxy-deoxyguanocine (8-OHdG) as oxidative stress marker. 8-OHdG was observed in the outer nuclear layer (ONL) and retinal pigment epithelium (RPE) during light exposure. In light-damaged retinas CuZn-SOD labeling was up regulated in the ONL and RPE. Mn-SOD labeling was up regulated in rod inner segments (RIS) during light exposure and that in the RPE was up regulated after exposure. GPX labeling was observed in rod outer segments (ROS) during light exposure. GPX labeling was also observed in the RPE during and after light exposure. All three enzymes were observed in the outer retina, which suffered light damage, but occurred in defferent layers except within the RPE, in which case all three were expressed. These enzymes may play complementary roles as protective factors in light-damaged retinas.

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Expression of Superoxide Dismutase Isoforms in Inflamed Gingiva (염증성 치은에서 superoxide dismutase isoform의 발현에 대한 연구)

  • Na, Hei-Jin;Kim, Ok-Su;Park, Byung-Ju
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.97-112
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    • 2006
  • 유리 라디칼과 활성 산소종, 산화방지제 간의 불균형이 염증성 구강내 질환의 발생과 진행에 있어 중요한 역할을 한다는 주장이 제기되었고 최근에는 만성 염증성 치주질환에서도 산화에 의한 소실이 관찰되었다. 다양한 내적인 항산화 방어 기전 중 superoxide dismutase 가 $O_2$$H_2O_2$로 효과적으로 전환시킴으로써 활성산소종에 대한 일차적인 방어를 맡고 있다. 현재까지 인간에서 발견된 superoxide dismutase 는 cytoplasmic copper-zinc SOD와 mitochondrial manganase SOD, extracellular SOD의 3가지 아형이다. 이번 연구는 만성 치주질환을 가전 환자의 치주조직에서 효소 항산화제인 SOD의 발현정도를 알아봄으로써 질환조직 내의 산화자극 정도를 평가해 보고자하였다. 전남대학교 치주과에 내원한 33명의 만성 치주질환자와 20명 의 임상적으로 건강한 대상으로부터 조직을 얻어 Cu/Zn-SOD와 Mn-SOD, EC-SOD를 이용한 면역조직화학 염색을 시행하였다. 임상적 소견과 조직학적 소견이 일치하지 않아 조직학적 소견을 기준으로 건강한 조직, 경도, 중등도, 중도 치주질환 조직으로 그룹을 나누고 완전한 상피와 결합조직을 가진 27개의 표본에 대한 분석을 시행하였다. 치주질환 조직에서 건강한 조직에 비해 Cu/Zn-SOD가 상피의 기저층과 상피에 근접한 결합조직에서 발현되고 Mn-SOD는 염증이 증가함에 따라 크게 상피의 과립증과 각화층, 그리고 상피에 근접한 결합조직에서 발현됨으로써 활성산소종이 치주조직 파괴에 관여한다는 것을 알 수 있었다. 세 아형 모두 혈관주위에서 발현되었고 특히 EC-SOD는 작은 모세혈관주위에서만 발현되었으나 염증에 의해 혈관벽이 두꺼워지고 혈관 수가 증가한 곳에서 뚜렷하게 염색되었다. 이번 연구는 염증성 치주조직내 증가된 SOD의 활성이 치주질환자의 산화자극 정도와 관련되어 있음을 시사하였다.

The Effect of Red Ginseng Extracts on the Superoxide Dismutase, Peroxidase and Catalase Activities in the Liver of Gamma Ray Irradiated Mice (홍삼 투여가 방사선에 조사된 생쥐 간세포의 Superoxide Dismutase, Peroxidase 및 Catalase 활성도에 미치는 영향)

  • Chun, Chul;Chang, Che-Chul
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.29-34
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    • 1993
  • This study was undertaken to investigate the effect of red ginseng extract (5.5 mg/mouse ip) on the activities of superoxide dismutase (SOD), peroxidase and catalase in the liver of the gamma ray irradiated male mice. The experimental groups consisted of control, red ginseng extract injection group, irradiation (8 $Gy^{60}$Co) group and red ginseng extract injection after irradiation group. in red ginseng extract injection group, SOD, peroxidase and catalase activities were similar to that in the control group. In irradiation group SOD, peroxidase and catalase activities increased progressively until the 2nd day after the treatment and then decreased thereafter, whereas red ginseng extract injection after irradiation group recovered more rapidly than irradiation group. The above results suggested that red ginseng extract injection after irradiation group have the recovery effects on the activities of SOD, peroxidase and catalase after radiation injury in the liver of male mice.

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Anaerobic Respiration of Superoxide Dismutase-Deficient Saccharomyces cerevisiae under Oxidative Stress

  • Lee, Sun-Mi;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.15-18
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    • 1998
  • The entanol productivity of superoxide dismutase (SOD)-deficient mutants of Saccharo-Myces cerevisiae was examined under the oxidative stress by Paraquat. It was observed that MnSOD-deficient mutant of S. cerevisiae had higher ethanol productivity than wild type or CuZnSOD-deficient yeast both in aerobic and in anaerobic culture condition. Pyruvated dehydrogenase activity decreased by 35% and alcohol dehydrogenase activity increased by 32% were observed in MnSOD-deficient yeast grown aerobically. When generating oxygen radicals by Paraquat, the ehanol productivity was increased by 40% in CuZnSOD-deficient or wild strain, resulting from increased activity of alcohol dehydrogenase and decreased a activity of pyruvate dehydrogenase. However, the addition of ascorbic acid with Paraquat returned the enzyme activities at the level of control. These results imply that SOD-deficiency in yeast strains may cause the metabolic flux to shift into anaerobic ethanol fermentation in order to avoid their oxidative damages by Paraquat.

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Characterization of an Iron- and Manganese-containing Superoxide Dismutase from Methylobacillus Sp. Strain SK1 DSM 8269

  • Seo, Sung Nam;Lee, Jae Ho;Kim, Young Min
    • Molecules and Cells
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    • v.23 no.3
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    • pp.370-378
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    • 2007
  • A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.