• Title/Summary/Keyword: Metallothionein

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The Characteristics of Fish Metallothionein and Its Application to the Biomonitoring for the Evaluation of Water Pollution (어류 Metallothionein의 툭성 및 수질오염 평가를 위한 생물모니터링에의 응용)

  • 황갑수
    • Environmental Analysis Health and Toxicology
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    • v.12 no.3_4
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    • pp.15-22
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    • 1997
  • This experiment was performed to examine the immuno-reactive characteristics of fish metal-binding protein, metallothionein (MT), and gain the practical understandings for the proposed use of fish MT as a biomarker. Liver MT induced by Cd in the silver carp was seperated and purified by gel filtration chromatography and ion exchange chromatography. The immuno-reactivity of fish MT was examined with 3 rabbit antisera. Fish MT showed little reactivity with rabbit anti-rat MT antiserum and a weak reactivity with anti-MT peptide antiserum while showed a strong reactivity with rabbit anti-fish MT antiserum. The time-course change of liver MT in the silver carp, after waterborne exposure to 1 ppm of Cd, was checked by Cd-hem method and established competitive ELISA. In both cases, the induction of liver MT showed a good increasing relationship with the exposure days. The results indicate that the fish MT can be developed as a useful biomonitoring means in the toxicological study and for the evaluation of water pollution.

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The Study on the Establishment of Specific ELISA for the Detection of Fish Metallothionein (어류 Metallothionein에 대한 특이적 면역 효소 측정법의 확립에 관한 연구)

  • 황갑수
    • Environmental Analysis Health and Toxicology
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    • v.11 no.1_2
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    • pp.11-17
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    • 1996
  • The purpose of this experiment is to estabilish the sensitive and specific ELISA (enzyme linked immunosorbent assay) system for the detection of fish metallothionein (MT). Silver carp were injected with CA of 1-8mg/kg body wt. 4 times during 10 days. Silver carp was very tolerant species to CA. Cd induced MT in liver was seperated and purified by gel filtration chromatography and ion exchange chromatography and identified by spectrophotometry, native gel electrophoresis and western blot analysis. The rabbit antiserum was produced by immunizing rabbit with lyophilized MT, and the competitive ELISA system was estabilished for the detection of fish MT. In the present ELISA system, the detection limit was about 33 ng/ml. When this ELISA system was employed to determine the MT level in the supernatant sample of fish liver homogenate, the reaction curve showed a good parallel corelationship with the calibration curve over a certain dilution range. The results indicate that the competitive ELISA can be a useful tool for the detection of fish MT in the toxicological study and the evaluation of water pollution.

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Metallothionein Induces Site-specific Cleavages in tRNAPhe

  • Seon, Jung-Yun;Koh, Moon-Joo
    • Bulletin of the Korean Chemical Society
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    • v.26 no.6
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    • pp.921-924
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    • 2005
  • It is known that metallothionein (MT) plays a role in the scavenging of free radicals, which is produced under various stress conditions. MT may function as an antioxidant that protects against oxidative damage of DNA, protein, and lipid induced by superoxide anion, hydrogen peroxide, hydroxyl radical, nitric oxide, and peroxynitrite. This study was undertaken to test the hypothesis that MT also protects from RNA damage induced by peroxynitrite, an important reactive nitrogen species that causes a diversity of pathological processes. A cell-free system was used. RNA damage was detected by the mobility of $tRNA^{Phe}$ in electrophoresis. Cleavages on tRNA were not induced by 3-morpholinosydnomine, which produces peroxynitrite directly. MT induced tRNA damage which was site specific.

Development of Antitoxic Agents from Korean Medicinal Plants. Part 8. -Effects of Methanol Fraction of Lonicerae flos on the Accumulation of Cadmium and Metallothionein in Rats.- (한국산 생약으로 부터 해독물질의 개발(제8보)-금은화 메탄올 분획이 흰쥐 장기내 카드뮴 축적과 Metallothionein 생성에 미치는 영향-)

  • 김영옥;이종섭;박경옥;한두석;유일수;곽정숙;백승화
    • Toxicological Research
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    • v.12 no.1
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    • pp.41-46
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    • 1996
  • This study was performed to investigate the antitoxic component in methanol fraction of Lonicerae fios. The results were as follows: 1. When a 500 ppm of water soluble fraction of Lonicerae flos was administered, it showed the highest antitoxic effect. 2. Generally, detoxication effects by methanol fraction of Lonicerae fios increased. When the ethyl acetate soluble fraction of Lonicerae rios was administered, it exhibited the highest antitoxic effect against the toxicity of cadmium in Liver. 3. The combined administration of cadmium and methanol fraction of Lonicerae rios significantly increased metallothionein in liver compared to administration of cadmium only. This phenomenon was more remarkable when the ethyl acetate soluble fraction of Lonicerae fios was administered with cadmium chloride.

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Purification of Carp Serum Metallothionein Induced in Carp with Cadmium and Production of Polyclonal Antibody (카드뮴 유도 잉어 혈액으로부터의 Metallothionein 정제 및 이에 대한 다클론 항체 생산)

  • Ryu, Hyung-Seok;Kang, Ho-Joon;Kim, Seul Ki;Kim, Namsoo;Kim, Woo-Yeon
    • Food Engineering Progress
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    • v.13 no.4
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    • pp.314-319
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    • 2009
  • Metallothioneins (MTs) are low molecular weight, cysteine-rich, and heavy metal binding proteins, which could be induced with heavy metals such as Cd, Hg, Zn and Cu in liver, kidney, and in cultured cells. By using ion exchange chromatography on DE-52, MT was purified from the serum of carp induced with cadmium in order to produce antibody against MT. Polyclonal antibody produced against purified carp MT reacted well with MT in the serum of carp induced with cadmium, whereas control serum did not. This may indicate that the polyclonal antibody against the carp MT could be used for the preparation of biosensors to detect MT in fishes like carp.

The Effect of Metallothionein on the Activity of Enzymes Invelved in Remival of Reactive Oxygen Species

  • Go, Mun Ju;Kim, Hui Jeong
    • Bulletin of the Korean Chemical Society
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    • v.22 no.4
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    • pp.362-366
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    • 2001
  • To show the effects of metallothionein (MT) on the activity of enzymes involved in the removal of reactive oxygen species, MT has been added to the assay systems of superoxide dismutase (SOD), catalase and peroxidase. We have used assay systems of SOD based on NADPH oxidation and nitrite formation from hydroxylammonium chloride as an assay of superoxide breakdown rate. The two assay systems showed different results at the high concentration of MT. MT showed the scavenging of superoxide in the SOD assay system in the presence and absence of SOD. MT added to the SOD assay system behaved as an activator of SOD, but apo-MT behaved as an inhibitor. When MT was added to the assay system in the presence of a fixed amount of SOD, the breakdown rate of superoxide increased. The effects of MT on the decomposition of hydrogen peroxide and the activity of catalase and peroxidase decomposing hydrogen peroxide were evaluated. MT decreased the activities of catalase and peroxidase. We have concluded that the function of MT as an antioxidant might effect the level of superoxide scavenging and not the level of hydrogen peroxide.

Effects of Water Extracts of Persimmon Leaves to Cadmium Toxicity in Rats by Inhalation Exposure (시엽 추출물이 카드뮴에 흡입폭로된 랫드의 독성에 미치는 영향)

  • Kang Sung Ho;Chun Byung Yeol;Kim Sang Duck;Song Young Son;Lee Ki Nam;Jeung Jae Yeal
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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    • pp.78-88
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    • 2002
  • Experimental animals were divided into 5 groups; normal, cadmium control, and 3 experimental groups. Cadmium control and experimental groups were exposed to 1 mg/㎥ of cadmium aerosol in air by inhalation exposure for 6 hours/day, 5 days/week during 4 weeks. Dosages of 20, 40, and 80mg/kg of extracts of persimmon leaves were intraperitoneally injected to experimental groups respectively and several toxicological parameters and induction of metallothionein were measured from the rats that inhaled cadmium aerosol in air. The results of this study were as follows. Cadmium concentration that cadmium control and experimental groups were inhaled was 0.980±0.061 mg/㎥. Mass median diameter of cadmium aerosol for inhalation exposure was 4.93±0.483㎛. Cadmium content of normal group in lung was 0.088㎍/g and the highest cadmium content in lung, 55.492㎍/g was from 80mg/kg dose group. Cadmium concentration of normal group in blood was 0.348㎍/100㎖ and the highest cadmium concentration in blood, 2.642㎍/100㎖ was from cadmium control. Cadmium concentration of normal group in liver was 0.010㎍/g and the highest cadmium concentration in liver, 31.100㎍/g was from 20mg/kg dose group. Cadmium concentration of normal group in kidney was 0.030㎍/g and the highest cadmium concentration in kidney, 2.526㎍/g was from cadmium control. Cadmium concentration of normal group in intestine was O.064㎍/g and the highest cadmium concentration in intestine, 0.300㎍/g was from 80mg/kg dose group. The highest cadmium concentration in urine by week was 6.080㎍/day from 20mg/kg dose group in the fouth week and the highest cadmium concentration in feces by week was 341.731㎍/day from 20mg/kg dose group in the fouth week. Metallothionein concentration of normal group in lung was 5.769㎍/g and the highest in lung, 30.986㎍/g was from 80mg/kg dose group. Metallothionein concentration of normal group in liver was 38.856㎍/g and the highest in liver, 169.378㎍/g was from 40mg/kg dose group. Metallothionein concentration of normal group in kidney was 22.228㎍/g and the highest in kidney, 47.898㎍/g was from 80mg/kg dose group. Metallothionein concentration of normal group in intestine was 2.170㎍/g and the highest in intestine, 13.642㎍/g was from 80mg dose group.

Metallothionein gene(pPMT)와 Manganese Transport Gene mntA(pZH3-5)를 포함한 재조합 Escherichia coli를 이용하여 수용액상에서의 Cadmium의 선택적 제거

  • Kim, Se-Gwon;Baek, Seung-Hak;Kim, Eun-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.426-429
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    • 2000
  • Recombinant E. coli JM109(pZH3-5/pMT) harboring manganese transport gene(mntA) and metal sequestering protein, metallothionein(MT), was cultivated to accumulate cadmium in aqueous phase. Bioaccumulation followed Michaelis-Menten type kinetics. Equilibrium isotherm showed Langmuir type isotherm. The optimum pH for $Cd^{2+}$ uptake was 7-7.5.

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MOLECULAR BIOMARKER OF CADMIUM EXPOSURE IN FRESHWATER FISH: SENSITIVITY AND SPECIFICITY

  • Park, Kwangsik;Heekyung Bae;Nam, Seong-Sook;Kim, Enkyoung
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.174-174
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    • 2002
  • Metallothioneins (MTs) are known to be induced by heavy metals in various organs of different species and represent a potential biomarker of aquatic contamination by heavy metals. In this work, cloning and sequencing of a metallothionein gene in crucian carp (Carassius auratus) was done and sensitivities and specificities of the gene expressions were compared.(omitted)

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