• 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.

• BMB Reports
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• 제30권1호
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• pp.60-65
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• 1997

Expression of human Cu.Zn-superoxide dismutase (SOD) with activity comparable to human erythrocyte enzyme was achieved in E. coli B21(DE3) by using the pET-17b expression vector containing a T7 promoter. Recombinant human SOD was found in the cytosol of disrupted bacterial cells and represented > 25% of the total bacterial proteins. The protein produced by the E. coli cells was purified using a combination of ammonium sulfate precipitation, Sephacryl S-100 gel filtration and DEAE-Sephacel ion exchange chromatography. The recombinant Cu,Zn-SOD and human erythrocyte enzyme were compared using dismutation activity, SDS-PAGE and immunoblotting analysis. The mass of the subunits was determined to be 15,809 by using a electrospray mass spectrometer. The copper specific chelator. diethyldithiocarbamate (DOC) reacted with the recombinant Cu,Zn-SOD. At $50{\mu}M$ and $100{\mu}M$ concentrations of DOC, the dismutation activity was not inhibited for one hour but gradually reduced after one hour. This result suggests that the reaction of DOC with the enzyme occurred in two distinct phases (phase I and phase II). During phase I of this reaction, one DOC reacted with the copper center, with retention of the dismutation activity while the second DOC displaced the copper, with a loss of activity in phase II.

• 한국자원식물학회지
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• 제12권3호
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• pp.234-239
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• 1999

두릅나무과 식물 6종의 잎으로부터 2개의 공통적인 superoxide dismutase(SOD) isoenzyme이 구분되었다. 이들 공통적인 isoenzyme(SOD 4와 SOD 6)의 패턴은 오갈피속 식물 중에서 민가시오갈피나무(A. senticosus for. inermis) 잎에서 가장 다양하였고, 그 활성은 섬오갈피나무(A. koreanum) 잎에서 가장 높았다. 그리고 SOD 4와 SOD 6은 $H_2O$$_2$와 KCN에 의한 선택적 저해로부터 각각 Fe-SOD와 CuZn-SOD인 것으로 밝혀졌다. 또한, SOD isoenzyme의 패턴은 섬오갈피나무의 성숙한 잎과 수피, 근피에서 차이가 없었으나 그 활성은 조직별로 차이가 나타났으며, Fe-SOD는 근피에서, CuZn-SOD은 잎에서 상대적으로 높게 나타났다. 그리고, CuZn-SOD나 Fe-SOD 모두 30-4$0^{\circ}C$에서 높은 활성 을 나타내었으나 그 이상의 온도에서는 활성이 저해되었다.

• 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.

• Journal of Plant Biotechnology
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• 제29권1호
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• pp.7-13
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• 2002

Superoxide dismutase (SOD)를 과실에서 고발현시킨 형질전환 토마토 (서광과 꼬꼬)를 개발하였다. 카사바 배양세포에서 분리한 CuZnSOD (mSOD1)를 과실에 우세적으로 발현하는 ascorbate oxidase promoter (ASOp)를 이용하여 ASOp :: mSOD1/pBI101 벡터를 제작한 후 Agrobacterium 매개로 자엽 절편체를 형질전환하였다. Kanamycin 저항성 식물체를 기관발생 경로로 재분화시킨 후 Southern 분석으로 형질전환을 확인하였다. 서광과 꼬꼬 토마토의 형질전환체와 대조구 식물체의 과실을 성숙 단계별로 분류하여 단백질 함량과 SOD 비활성도 (units/mg protein)를 측정한 결과, 단백질 함량은 열매가 익은 단계로 갈수록 점점 감소하여 완전히 익은 단계에서 가장 낮았다. SOD 비활성도는 형질전환 토마토의 열매의 모든 단계에서 대조구보다 높았으며 완전히 성숙한 과실에서 가장 높았다. 성숙한 형질전환 서광과 꼬꼬 과실에서 SOD 비활성도는 비형질전환의 것보다 각각 약 1.6배와 약 2.2배 높았다. SOD isoenzyme gel 분석에서 도입한 mSOD1로 추정되는 CuZnSOD 밴드가 형질전환체에서 과실 성숙에 따라 강하게 발현되었다. 이상의 결과로서 ASO promoter에 의해 SOD 유전자가 토마토 과실에 특이적으로 발현됨이 확인되었다.

• Journal of Periodontal and Implant Science
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• 제36권1호
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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 $O_2$를 $H_2O_2$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.

• 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.

• Fisheries and Aquatic Sciences
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• 제12권3호
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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.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.630-644
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• 2003

A human Cu, Zn-superoxide dismutase (Cu, Zn-SOD) was fused with a Tat PTD of HIV-1 to produce a novel anti-aging ingredient, Tat-SOD for cosmeceuticals. Test of stability and evaluation of transduction efficacy and enzymatic activity suggest Tat-SOD is an effective active ingredient for anti-aging treatment.

• Parasites, Hosts and Diseases
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• 제29권3호
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• pp.259-266
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• 1991

폐흡충 성충의 인산완충액 추출액을 원심분리하여 만든 세포질 현탁액을 조효소(조효소)로 사용하여 Xanthine- zanthine oxidase system으로 superoxide digmutase 환성을 측정한 결과 비활성도(비활성도: specific activity)는 4.3 units/mg이었다. 이 효소의 활성도를 30∼85% ammonium sulfate 침전 및 DEAE-Trisacryl M anion exchange chromatography와 Sephadex G-100 column chromatography를 통과시키면서 측정하는 방법으로 superoxide dismutase를 정제하고, 정제한 효소의 생화학적 특성을 관찰하여 다음과 같은 결과를 얻었다. 1. 정제과정을 통해 세포질 내 superoxide dismutase를 150배 정제하였다. Sephadex G-100 gel filtration에 의해 계산한 이 효소의 분자량은 34kDa이었고 환원성 SDS-PAGE상 subunit가 17 kDa이었다. 따라서 이 효소는 subunit 두개로 구성된 중항체로 판단하였다. 3. 이 효소는 1 mM 이상의 cyanide 농도에서는 효소 황성이 100% 억제되었고, 5mM 농도의 azide에서는 11.1%, 10mM azide에서는 22.2% 그 환성이 각각 억제되었다. 3. 이 효소는 완충액의 pH가 10.0일 때 효소 황성이 5.4배 증가되었다. 이상의 결과로 폐흡충의 세포질 내에 존재하는 superoxide dismutase는 구리와 아연을 함유한 superoxide dismutase라고 판단할 수 있었다.