• Title/Summary/Keyword: Glutathione Redox

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An L-Type Thioltransferase from Arabidopsis thaliana Leaves

  • Kim, Tae-Soo;Cho, Young-Wook;Kim, Joon-Chul;Jin, Chang-Duck;Han, Tae-Jin;Park, Soo-Sun;Lim, Chang-Jin
    • BMB Reports
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    • v.32 no.6
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    • pp.605-609
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    • 1999
  • Thioltransferase, also called glutaredoxin, is a general GSH-disulfide reductase of importance for redox regulation. Previously, the protein thioltransferase, now called S-type thioltransferase, was purified and characterized from Arabidopsis thaliana seed. In the present study, a second thioltransferase, called L-type thioltransferase, was purified to homogeneity from Arabidopsis thaliana leaves. The purification procedures included DEAE-cellulose ion-exchange chromatography, Sephadex G-50 gel filtration, and glutathione-agarose affinity chromatography. The purified enzyme was confirmed to show a unique band on SDS-PAGE and its molecular weight was estimated to be 26.6 kDa, which appeared to be atypical compared with those of most other thioltransferase. It could utilize 2-hydroxyethyl disulfide, S-sulfocysteine, and insulin as substrates, and also contained dehydroascorbate reductase activity. Its optimum pH was 8.5 and its activity was greatly activated by L-cysteine. When it was kept for 30 min, it appeared to be very stable up to $70^{\circ}C$. It was activated by $MgCl_2$ and, on the contrary, inhibited by $ZnCl_2$, $MnCl_2$, and $AlCl_3$.

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The Chemopreventive Effects of Antioxidant Enzyme (항산화효소의 암 예방 효과 및 발암 억제 기전)

  • Jung Hwa-Jin;Choi Yoon-Joo;Won Chang-Won;Seo Young-Rok
    • Environmental Mutagens and Carcinogens
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    • v.26 no.2
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    • pp.45-47
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    • 2006
  • The reactive oxygen species (ROS) caused the damage of macro molecules, many degenerative disease and cancer, which was produced in process of the aerotropic metabolic pathway as well as in response to the various genotoxic stresses. Recently, redox systems including the number of antioxidant proteins such as catalase, glutathione peroxidase, heam-containing peroxidase, peroxiredoxin and superoxide dismutase (SOD) has been reported to have chemopreventive effects. Antioxidant proteins has been known to have the activity directly removing ROS and affecting the protein-protein interaction and cell signaling to induce the cellular responses. We need to understand the mechanism of antioxidants prevent DNA damage from oxidative stresses for researching the cancer prevention and providing the development of cancer therapeutic drug.

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Scavenging Reactive Oxygen Species by Rice Dehydroascorbate Reductase Alleviates Oxidative Stresses in Escherichia coli

  • Shin, Sun-Young;Kim, Il-Sup;Kim, Yul-Ho;Park, Hyang-Mi;Lee, Jang-Yong;Kang, Hong-Gyu;Yoon, Ho-Sung
    • Molecules and Cells
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    • v.26 no.6
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    • pp.616-620
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    • 2008
  • Maintaining redox balance is one of the crucial requirements for a cell to endure stress from the outside. Dehydroascorbate reductase (DHAR; EC 1.8.5.1) plays an important role in the ascorbate-glutathione cycle; one of the major ROS scavenging systems in most known biological systems. A cDNA clone of the DHAR gene from Oryza sativa (OsDHAR) was isolated and overexpressed in Escherichia coli BL21 (DE3) strain from the pET-28a(+) expression vector. The OsDHAR transformed E. coli cells showed significantly higher DHAR activity and a lower level of ROS than the E. coli cells transformed by an empty pET-28a(+) vector. Also, the DHAR-overexpressing E. coli strain was more tolerant to oxidant- and heavy metal-mediated stress conditions than the control E. coli strain. The results suggest that the overexpressed rice DHAR gene effectively functions in a prokaryotic system and provide protection to various oxidative stresses.

Hydrogen Peroxide Induces Apoptosis of BJAB Cells Due to Formation of Hydroxyl Radicals Via Intracellular Iron-mediated Fenton Chemistry in Glucose Oxidase-mediated Oxidative Stress

  • Lee, Jeong-Chae;Son, Young-Ok;Choi, Ki-Choon;Jang, Yong-Suk
    • Molecules and Cells
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    • v.22 no.1
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    • pp.21-29
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    • 2006
  • The aim of this study was to determine if hydrogen peroxide ($H_2O_2$) generated by glucose oxidase (GO) induces apoptosis or necrosis of BJAB cells and which radical is the direct mediator of cell death. We found that GO produced $H_2O_2$ continuously in low concentrations, similar to in vivo conditions, and decreased proliferation and cell viability in a dose-dependent manner. The GO-mediated cytotoxicity resulted from apoptosis, and was confirmed by monitoring the cells after H33342/Annexin V/propidium iodide staining. Decreases of mitochondrial membrane potential and intracellular glutathione level were found to be critical events in the $H_2O_2$-mediated apoptosis. Additional experiments revealed that $H_2O_2$ exerted its apoptotic action through the formation of hydroxyl radicals via the Fenton rather than the Haber-Weiss reaction. Moreover, intracellular redox-active iron, but not copper, participated in the $H_2O_2$-mediated apoptosis.

Effects of Heterologous Expression of Thioredoxin Reductase on the Level of Reactive Oxygen Species in COS-7 Cells

  • Kang, Hyun-Jung;Hong, Sung-Min;Kim, Byung-Chul;Park, Eun-Hee;Ahn, Kisup;Lim, Chang-Jin
    • Molecules and Cells
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    • v.22 no.1
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    • pp.113-118
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    • 2006
  • Thioredoxin reductase (TrxR), a component of the redox control system involving thioredoxin (Trx), is implicated in defense against oxidative stress, control of cell growth and proliferation, and regulation of apoptosis. In the present study a stable transfectant was made by introducing the vector pcDNA3.0 harboring the fission yeast TrxR gene into COS-7 African green monkey kidney fibroblast cells. The exogenous TrxR gene led to an increase in TrxR activity of up to 3.2-fold but did not affect glutathione (GSH) content, or glutaredoxin and caspase-3 activities. Levels of reactive oxygen species (ROS), but not those of nitric oxide (NO), were reduced. Conversely, 1-chloro-2,4-dinitrobezene (CDNB), an irreversible inhibitor of mammalian TrxR, enhanced ROS levels in the COS-7 cells. After treatment with hydrogen peroxide, the level of intracellular ROS was lower in the transfectants than in the vector control cells. These results confirm that TrxR is a crucial determinant of the level of cellular ROS during oxidative stress as well as in the normal state.

Effect of Selenium on the Thyroid gland Antioxidative Metabolisms in Rat Model by Ionizing Radiation (셀레늄이 전리방사선에 의한 힌쥐 모델에서의 갑상선 항산화계에 미치는 영향)

  • Choi, Hyung-Seok;Choi, Jun-Hyeok;Jung, Do-Young;Kim, Jang-Oh;Shin, Ji-Hye;Min, Byung-In
    • Journal of radiological science and technology
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    • v.40 no.1
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    • pp.135-142
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    • 2017
  • Selenium (Se), which is natural materials existing was known as an important component of selenoprotein, one of the important proteins responsible for the redox pump of a living body. Selenium was orally administered to Rat and irradiated with 10 Gy of radiation. Then, the thyroid gland was used as a target organ for 1 day, 7 days and 21 days to investigate the radiation protection effect of selenium (Se) through changes of blood components, thyroid hormones (T3, T4), antioxidant enzyme (GPx) activity and thyroid tissue changes. As a result, there was a significant protective effect of hematopoietic immune system(hemoglobin concentration, neutrophil, platelet)(p<0.05). The activity of Glutathione Peroxidase (GPx), the antioxidant enzyme, and the activity of the target organ, thyroid hormone (T3, T4), also showed significant activity changes (p<0.05). In the observation of tissue changes, it was confirmed that there was a protective effect of thyroid cell damage which caused the cell necrosis by radiation treatment. Therefore, it is considered that selenium(Se) can be utilized as a radiation defense agent by inducing immunogenic activity effect of a living body.

Antioxidant Enzyme Activity and Anti-Adipogenic Effects of (-)-Epigallocatechin-3-Gallate in 3T3-L1 Cells ((-)-Epigallocatechin-3-Gallate의 3T3-L1 세포에서 항산화 효소 활성 및 지방세포 분화 억제 효과)

  • Kim, Younghwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1293-1299
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    • 2017
  • Obesity contributes to the development of diseases, such as type II diabetes, hypertension, coronary heart disease, and cancer. In addition, oxidative stress caused by reactive oxygen species (ROS) is recognized widely as a contributing factor in the development of chronic diseases. This study was examined the antioxidant and anti-adipogenic activities of epigallocatechin-3-gallate (EGCG) in 3T3-L1 preadipocytes. 3T3-L1 cells were differentiated with or without EGCG for 6 days. The production of glutathione (GSH) and the activities of the antioxidant enzymes, such as glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) were measured. EGCG inhibited significantly the lipid accumulation and the expression of adipogenic specific proteins including CCAAT/enhancer binding protein ${\alpha}$ and adipocyte fatty acid binding protein. The production of intracellular ROS was decreased significantly by EGCG in 3T3-L1 cells. EGCG increased the GSH production and the activities of GPx, GR, CAT, and SOD. Moreover, EGCG increased the protein expression of glutamate-cysteine ligase and heme oxygenase-1 in 3T3-L1 cells. These results suggest that EGCG increased the activity and expression of antioxidant enzymes and suppressed the lipid accumulation in 3T3-L1 cells. Therefore, the use of phytochemicals that can maintain the GSH redox balance in adipose tissue could be promising for reducing obesity.

The Effect of Dietary Modified Potato Starch By Chemically Denatured Treatment and Potato Starch on the Weight Loss, Lipid Metabolism and Redox Antioxidant System in High Fat Diet-Induced Obese Rats (화학적 변성 및 생감자 전분이 고지방식이로부터 유도된 비만 흰쥐의 지질대사 및 항산화계에 미치는 영향)

  • Park, Soo-Jin;Choi, Mi-Kyeong;Kim, Jin-Suk;Lim, Hak-Tea;Kang, Myung-Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1251-1257
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    • 2008
  • For the first 42 days, we made rats obese by feeding them potato starch instead of corn starch and after that we fed them transformed potato starch by 4 groups for 70 days. The 4 groups are GPS group, SPS group, EZ group and H40 groups and each were fed normal potato, small potato, enzyme treated potato, and acid treated potato starches, respectively. We determined body weight and feeding efficiency, lipid profiles in serum, lipid peroxidation in tissues and redox antioxidant system as GSH and GP-x in vivo. As a result, there was no difference in the increment of body weight in the groups of GPS, EZ and H40. Therefore EZ group showed lower body weight increment than other groups. While GPS group and SPS group did not show significant difference in blood glucose, cholesterol level, LDL-cholesterol and TC, and their measured values were lower than those of EZ and H40 groups. No significant difference was found in HDL-cholesterol level except for GPS group. Furthermore, when calculating atherogenic index (AI) by HDL-cholesterol and TC contents, H40 group showed higher measured value than other groups. When measuring the lipid peroxidation in serum, kidney and liver tissues, the serum lipid peroxidation in H40 group was higher than others. In the tissue of liver and kidney, EZ and H40 groups showed significantly lower contents than others. The content of GSH showed different tendency in each tissue, but the measured value of GP-x activity was lower in SPS group.

Mountain-cultivated ginseng protects against cognitive impairments in aged GPx-1 knockout mice via activation of Nrf2/ChAT/ERK signaling pathway

  • Bao Trong Nguyen;Eun-Joo Shin;Ji Hoon Jeong;Naveen Sharma;Ngoc Kim Cuong Tran;Yen Nhi Doan Nguyen;Dae-Joong Kim;Myung Bok Wie;Yi Lee;Jae Kyung Byun;Sung Kwon Ko;Seung-Yeol Nah;Hyoung-Chun Kim
    • Journal of Ginseng Research
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    • v.47 no.4
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    • pp.561-571
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    • 2023
  • Background: Escalating evidence shows that ginseng possesses an antiaging potential with cognitive enhancing activity. As mountain cultivated ginseng (MCG) is cultivated without agricultural chemicals, MCG has emerged as a popular herb medicine. However, little is known about the MCG-mediated pharmacological mechanism on brain aging. Methods: As we demonstrated that glutathione peroxidase (GPx) is important for enhancing memory function in the animal model of aging, we investigated the role of MCG as a GPx inducer using GPx-1 (a major type of GPx) knockout (KO) mice. We assessed whether MCG modulates redox and cholinergic parameters, and memory function in aged GPx-1 knockout KOmice. Results: Redox burden of aged GPx-1 KO mice was more evident than that of aged wild-type (WT) mice. Alteration of Nrf2 DNA binding activity appeared to be more evident than that of NFκB DNA binding activity in aged GPx-1 KO mice. Alteration in choline acetyltransferase (ChAT) activity was more evident than that in acetylcholine esterase activity. MCG significantly attenuated reductions in Nrf2 system and ChAT level. MCG significantly enhanced the co-localization of Nrf2-immunoreactivity and ChAT-immunoreactivity in the same cell population. Nrf2 inhibitor brusatol significantly counteracted MCG-mediated up-regulation in ChAT level and ChAT inhibition (by k252a) significantly reduced ERK phosphorylation by MCG, suggesting that MCG might require signal cascade of Nrf2/ChAT/ERK to enhance cognition. Conclusion: GPx-1 depletion might be a prerequisite for cognitive impairment in aged animals. MCG-mediated cognition enhancement might be associated with the activations of Nrf2, ChAT, and ERK signaling cascade.

Expression of Cu/Zn SOD according to H2O2 in Hepatoma cell line (Hepatoma 세포주에서 H2O2 처리에 의한 Cu/Zn SOD의 발현)

  • Kim, Young-Min;Seo, Won-Sook
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.230-234
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    • 2007
  • Oxygen is required for many important aerobic cellular reactions, it may undergo electrontransfer reactions, which generate highly reactive membrane-toxic intermediates (reactive oxygen species, ROS), such as hydrogen peroxide, singlet oxygen, superoxide radical, hydroxyl radical, hydroperoxyl radical, hydroxy ion. Various mechanisms are available to protect cells against damage caused by oxidative free radicals, including scavenging enzyme systems such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). This antioxidant defense system is a very complex and finely tuned system consisting of enzymes capable of detoxifying oxygen radicals as well as low molecular weight antioxidants. In addition, repair and turnover processes help to minimize subcellular damage resulting from free radical attack. $H_2O_2$,one of the major ROS, is produced at a high rate as a product of normal aerobic metabolism. The primary cellular enzymatic defense systems against $H_2O_2$ are the glutathione redox cycle and catalase. From Northern blot analysis of total RNAs from cultured cell with $H_2O_2$ treatment, various results were obtained. Expression of Cu/Zn SOD decreased when cell passage increased, but the level of the Cu/Zn SOD was scarcely expressed in 35 passage.