• 제목/요약/키워드: Cu,Zn-Superoxide Dismutase

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들깨잎에 함유된 Superoxide Dismutase의 특성 및 항산화 활성 검정 (Test of Superoxide Dismutase Characteristics and Antioxidant Activity in Perilla Leaves)

  • 정일민;윤성중;김정태;곽재균;성재덕;서형수
    • 한국작물학회지
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    • 제40권4호
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    • pp.504-511
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    • 1995
  • SOD(superoxide dismutase)는 유해 활성산소에 의한 생리적 장해를 방지하는 방어기작의 한 구성요소이다. 들깨잎 SOD의 특성을 알아보기 위한 NBT(nitro blue tetrazolium) 환원법 을 이용하여 들깨와 자소잎의 SOD동위효소의 종류와 활성 및 이들의 항산화 활성을 Fe$^{2+}$/ascorbate, Fe$^{3+}$ -ADP/NADPH첨가제를 처리하여 조사하였다. 들깨잎에는 품종에 따라 3~4개의 주요 SOD동위효소가 있었다. SOD는 함유하고 있는 금속조효소에 따라 3종류로 분류되는데, 들깨잎에는 두개의 Cu/ZnSOD와 두개의 FeSOD을 함유하고 있었으나, 자소잎은 단지 Cu/ZnSOD만을 함유하고 있었다. Cu/ZnSOD는 들깨 품종에 따른 차이가 없었으나, FeSOD는 분자량이 다른 두개의 FeSOD동위효소의 존재 양상이 품종에 따라 차이를 나타내었으며, SOD활성 및 항산화 활성도 품종에 따라 비교적 큰 차이를 보였다. 비효소적인 Fe$^{2+}$/ascorbate첨가 및 Fe$^{3+}$ -ADP/NADPH첨가에 의해 유도된 지질 과산화의 억제에 미치는 영향을 조사한 결과 공시재료중 밀양 2호가 가장 강한 항산화 활성을 가지고 있는 것으로 확인되었다. 자소 잎의 경우는 SOD활성과 유사하게 항산화 활성 정도 가장 낮아서 들깨잎과는 뚜렷 이 구별되었다.

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DNA Cleavage Induced by the Reaction of Salsolinol with Cu,Zn-Superoxide Dismutase

  • Kang, Jung-Hoon
    • Bulletin of the Korean Chemical Society
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    • 제28권12호
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    • pp.2329-2332
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    • 2007
  • Salsolinol, endogenous neurotoxin, is known to be involved in the pathogenesis of Parkinson's disease (PD). In the present study, we have investigated the oxidative damage of DNA induced by the reaction of salsolinol with Cu,Zn-SOD. When plasmid DNA incubated with salsolinol and Cu,Zn-SOD, DNA cleavage was proportional to the concentrations of salsolinol and Cu,Zn-SOD. The salsolinol/Cu,Zn-SOD system-mediated DNA cleavage was significantly inhibited by radical scavengers such as mannitol, ethanol and thiourea. These results indicated that free radicals might participate in DNA cleavage by the salsolinol/Cu,Zn-SOD system. Spectrophotometric study using a thiobarbituric acid showed that hydroxyl radical formation was proportional to the concentration of salsolinol and was inhibited by radical scavengers. These results indicated that hydroxyl radical generated in the reaction of salsolinol with Cu,Zn-SOD was implicated in the DNA cleavage. Catalase and copper chelators inhibited DNA cleavage and the production of hydroxyl radicals. These results suggest that DNA cleavage is mediated in the reaction of salsolinol with Cu,Zn-SOD via the generation of hydroxyl radical by a combination of the oxidation reaction of salsolinol and Fenton-like reaction of free copper ions released from oxidatively damaged SOD.

Expression of a Cu-Zn Superoxide Dismutase Gene in Response to Stresses and Phytohormones in Rehmannia Glutinosa

  • Park, Myoung-Ryoul;Ryu, Sang-Soo;Yoo, Nam-Hee;Yu, Chang-Yeon;Yun, Song-Joong
    • 한국약용작물학회지
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    • 제13권5호
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    • pp.270-275
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    • 2005
  • Superoxide dismutases (SOD) are metalloenzymes that convert $O_2^-\;to\;H_2O_2$. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and $GA_3$. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

Cu/Zn Superoxide Dismutase 유전자 발현 운동신경세포주에서 NO 독성에 대한 Testosterone의 보호효과 (Testosterone-mediated Neuroprotection in NO Induced Cell Death of Motor Neuron Cells Expressing Wild Type or Mutant Cu/Zn Superoxide Dismutase)

  • 김남희;김현정;김만호;박경석;이광우
    • Annals of Clinical Neurophysiology
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    • 제8권1호
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    • pp.63-70
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    • 2006
  • Background: Testosterone is reported to have neuroprotective effect in various neurological diseases. Recently, the mechanism involved in nitric oxide (NO)-mediated motor neuron death is under extensive investigation. The Cu/Zn-superoxide dismutase (SOD1) mutations has been implicated in selective motor neuron death of amyotrophic lateral sclerosis (ALS) and it is said to play an important role in NO-mediated motor neuron death. However, neuroprotective effect of testosterone on motor neuron exposed to NO has rarely been studied. Methods: Motor neuron-neuroblastoma hybrid cells expressing wild-type or mutant (G93A or A4V) SOD gene were treated with $200{\mu}M$ S-nitrosoglutathione. After 24 hr, cell viability was measured by MTT assay. To see the neuroprotective effect of testosterone, pretreatment with 1 nM testosterone was done 1 hr before S-nitroglutathione treatment. To study the mechanism of protective effect, $20{\mu}M$ flutamide (androgen receptor antagonist) was also pretreated with testosterone 1 hr before S-nitroglutathione treatment. Results: S-nitrosoglutathione showed significant neurotoxic effect in all three cell lines. Percentage of cell death was somewhat different in each cell line. 1 nM testosterone showed neuroprotective effect in G93A and wild-type cell line. In A4V cell line, testosterone did not showed neuroprotective effect. The neuroprotective effect of testosterone was reversed by $20{\mu}M$ flutamide. Conclusions: These results indicate that testosterone induces neuroprotection in NO-mediated motor neuron death directly through the androgen receptor. This neuroprotective effect of testosterone varies according to the types of SOD1 gene mutation. These data suggest that testosterone may be of therapeutic value against ALS.

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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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    • 제9권2호
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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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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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    • 제3권1호
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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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Inactivation of Photosystem I in Cucumber Leaves Exposed to Paraquat-Induced Oxidative Stress

  • Park, Sun-Mi;Suh, Key-Hong;Kim, Jae-sung;Park, Youn-Il
    • Journal of Photoscience
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    • 제8권1호
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    • pp.13-17
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    • 2001
  • Cucumber leaves subjected to light chilling stress exhibit a preferential inactivation of photosystem(PS) I relative to PSII, resulting in the photoinhibition of photosynthesis. In light chilled cucumber leaves, Cu/Zn-Superoxide dismutase(SOD) is regarded as a primary target of the light chilling stress and its inactivation is closely related to the increased production of reactive oxygen species. In the present study, we further explored that inactivation of PSI in cucumber leaves is not a light chilling specific, but general to various oxidative stresses. Oxidative stress in cucumber leaves was induced by treatment of methylviologen(MV), a producer of reactive oxygen species in chloroplasts. MV treatment decreased the maximal photosynthetic O$_2$ evolution, resulting in the photoinhibition of photosynthesis. The photoinhibition of photosynthesis was attributable to the decline in PSI functionality determined in vivo by monitoring absorption changes around 820 nm. In addition, MV treatment inactivated both antioxidant enzymes Cu-Zn-superoxide dismutase and ascorbate peroxidase known sensitive to reactive oxygen species. From these results, we suggest that chloroplast antioxidant enzymes are the primary targets of photooxidative stress, followed by subsequent inactivation of PSI.

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Expression of Lily Chloroplastic Cu,Zn Superoxide Dismutase Enhances Resistance to Erwinia carotovora in Potatoes

  • Kim, Mi-Sun;Kim, Hyun-Soon;Kim, Yoon-Sik;Baek, Kwang-Hyun;Moon, Jae-Sun;Choi, Do-Il;Joung, Hyouk;Jeon, Jae-Heung
    • The Plant Pathology Journal
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    • 제23권4호
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    • pp.300-307
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    • 2007
  • Previously, a chloroplast-localized Cu,Zn superoxide dismutase (chCu,ZnSOD) was isolated from lily and the sense- and antisense- sequences of the lily chCu,ZnSOD were used to transform potato plants. Two selected lines, the sense- and anti-sense strand of transgenic plants, were further characterized for resistance to Erwinia carotovora, which is a severe pathogen affecting potato plants. Only the sense-strand transgenic potato, which contained less $O_2^{.-}$ and more $H_2O_2$ than wild-type and antisense-strand transgenic plants, showed increased resistance to E. carotovora. Additional studies using $O_2^{.-}$ or $H_2O_2$ scavengers in wild-type, sense-strand, and antisense-strand transgenic plants suggest that resistance to E. carotovora is induced by reduced $O_2^{.-}$ and is not influenced by $H_2O_2$. To the best of our knowledge, this report is the first study suggesting that resistance to E. carotovora is enhanced by reduced $O_2^{.-}$, and not by increased amounts of $H_2O_2$.

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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    • 제19권3호
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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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노화촉진 생쥐에서 노화에 따른 활성산소 생성 및 항산화능의 변화 (Age-associated Alteration in the Hapatic Superoxide Generation and Antioxidant Activities in the Senescence-accelerated Mice)

  • 정해영;김윤경
    • 약학회지
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    • 제36권5호
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    • pp.460-468
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    • 1992
  • Several bichemical parameters related to free radicals were estimated in senile-prone (P) and resistant(R) strains of male senescence-accelerated mice(SAM) at 2, 5 and 11 months of age. The superoxide generation was increased with age in SAM-R/1 and SAM-P/2. Compared to SAM-R/1, more generation of superoxide was significantly noted in the SAM-P/2 liver. The activities of Cu/Zn-superoxide dismutase and catalase were decreased during aging and these activities in SAM-P/2 were significantly lower than in SAM-R/l liver. The activities of glutathione S-transferase were varied with aging, whereas SAM-P/2 showed lower levels compared to SAM-R/l. The gradual decreases of glutathione, protein bound-SH and nonprotein bound-SH contents were noted with increasing age. SAM-P/2 liver contained lesser amounts of glutathione and nonprotein bound-SH compared to SAM-R. In conclusion, superoxide generation was increased whereas the antioxidant enzyme activities were decreased during aging in SAM-R/1. In addition, SAM-P/2 strain showed more superoxide generation and less antioxidant enzyme activities than SAM-R/1 in the liver, thus we assume that these factors might accelerate the senescence of SAM-P/2 strain.

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