• Title/Summary/Keyword: Isozymes

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Phospholipase D isozymes mediate epigallocatechin gallate-induced cyclooxygenase-2 expression in astrocyte cells

  • Kim, Shi-Yeon;Min, Kyoung-Jin;Joe, Eun-Hye;Min, Do-Sik
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.74-79
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    • 2004
  • Little is known about the effect of epigallocatechin-3 gallate (ESCG), a major constituent of green tea, on the expression of cyclooxygenase (COX)-2. Here, we studied the role of phospholipase D (PLD) isozymes in EGCG-induced COX-2 expression. Stimulation of human astrocytoma cells (U87) with EGCG induced formation of phosphatidylbutanol, a specific product of PLD activity, and synthesis of COX-2protein and its product, prostaglandin $E_2$ ($PGE_2$). Pretreatment of cells with 1-butanol, but not 3-butanol, suppressed EGCG-induced COX-2 expression and $PGE_2$ synthesis. Furthermore, evidence that PLD was involved in EGCG-induced COX-2 expression w3s provided by the observations that COX-2 expression was stimulated by over-expression of PLD1 or PLD2 isozymes and treatment with phosphatidic acid(PA), and that prevention of PA dephosphorylation by 1-propranolol significantly potentiated COX-2expression Induced by EGCG. EGCG induced activation of p38 mitogen-activated protein kinase (p38MAPK), and specific Inhibition of p38 MAPK dramatically abolished EGCG-Induced PLD activation, COX-2 expression, and $PGE_2$ formation. Moreover, protein kinase C (PKC) inhibition suppressed EGCG-induced p38 MAPK activation, COX-2 expression, and $PGE_2$ accumulation. The same pathways as those obtained in the astrocytoma cells were active in primary rat astrocytes, suggesting the relevance of the findings. Collectively, our results demonstrate for the first time that PLD isozymes mediate EGCG-induced COX-2 expression through PKC and p38 in immortalized astroglial line and normal astrocyte cells.

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Isozyme Variants in Genus Pinus by Simulated Acidic Rain (인공 산성비가 소나무속의 동위효소 양상에 미치는 영향)

  • EuiSooYoon
    • Korean Journal of Plant Resources
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    • v.10 no.4
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    • pp.305-313
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    • 1997
  • The effect of acid rain on plant isozyme response was studied with the treatment of simulated acid rain to Pinus densiflora, P. nigida and P. koraiensis for 9 months. The isozyme pattern of $\alpha$-Esterase($\alpha$-Est), Peroxidase(POD) and Glutamate dehydrogenase(GDH) were observed in the control (pH 5.6) and simulated acid rain (pH 3.5) treatment. No changes of isozyme pattern in $\alpha$-Est was observed in P. densiflora after 9 month treatment of simulated acid rain, but, two new isozymes were activated in P. nigida in the same treatment. In P. koraiensis, two new isozyme were activated but five isozymes were not activated. P. densiflora did not show any difference in POD and GDH after the treatment of simulated acid rain. P. nigida showed activation of eight and two isozymes in POD and GDH, respectively. P. koraiensis showed inactivation of 4 isozymes in POD but showed no changes In GDH.

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Dynamic Studies on Physiology and Biochemistry in American Ginseng Seed During Stratification Part III. POD Activity, Contents of DNA and RNA, Isozymes of POD and ES - (충적과정중 미국인삼 종자의 생리 및 생화하적 지표에 관한 동태학적 연구 III)

  • Huang, Yao-Ge;Li, Xiang-Gao;Kuang, Ya-Lan;Yan, Jie-Kun;Cui, Shu-Yu;Yu, Wen-Bo;Yang, Ji-Xiang;Liu, Ren-Song;Kim, Hack-Seang
    • Journal of Ginseng Research
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    • v.21 no.1
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    • pp.43-48
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    • 1997
  • Dynamic parameters of biochemistry including the POD (peroxidase) activity, contents of DNA and RNA, isozymes of POD and ES (esterase) in American send (Panax quinquefolium L.) seed are reported in the present paper. The dynamic changes of POD activity proved that the PAS (physiological afterripening stage) is a stage in which some substances are prepared for seed germination. The POD activity correlated with ER (embryo ratio) significantly DNA content changed little only within 0.0036∼0.013 mg/ml, which did not correlate with ER. RNA content changed from 0.1539 to 1.0313 mg/ml and correlated significantly with RE during all of the embryo afterripening. None of the POD isozyme band was obtained in ESGS (embryo slowly growth stage), but five bands in ERGS (embryo rapidly growth stage) and six bands in PAS. Four bands of ES isozymes were obtained in ESGS, but six bands in ERGS, particularly, the content of ES isozymes increased in PAS. All of these may provide some information for understanding the dormancy mechanisms of American song seed.

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Arabinoxylo- and Arabino-Oligosaccharides-Specific α-ʟ-Arabinofuranosidase GH51 Isozymes from the Amylolytic Yeast Saccharomycopsis fibuligera

  • Park, Tae Hyeon;Choi, Chang-Yun;Kim, Hyeon Jin;Song, Jeong-Rok;Park, Damee;Kang, Hyun Ah;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.272-279
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    • 2021
  • Two genes encoding probable α-ʟ-arabinofuranosidase (E.C. 3.2.1.55) isozymes (ABFs) with 92.3% amino acid sequence identity, ABF51A and ABF51B, were found from chromosomes 3 and 5 of Saccharomycopsis fibuligera KJJ81, an amylolytic yeast isolated from Korean wheat-based nuruk, respectively. Each open reading frame consists of 1,551 nucleotides and encodes a protein of 517 amino acids with the molecular mass of approximately 59 kDa. These isozymes share approximately 49% amino acid sequence identity with eukaryotic ABFs from filamentous fungi. The corresponding genes were cloned, functionally expressed, and purified from Escherichia coli. SfABF51A and SfABF51B showed the highest activities on p-nitrophenyl arabinofuranoside at 40~45℃ and pH 7.0 in sodium phosphate buffer and at 50℃ and pH 6.0 in sodium acetate buffer, respectively. These exoacting enzymes belonging to the glycoside hydrolase (GH) family 51 could hydrolyze arabinoxylo-oligosaccharides (AXOS) and arabino-oligosaccharides (AOS) to produce only ʟ-arabinose, whereas they could hardly degrade any polymeric substrates including arabinans and arabinoxylans. The detailed product analyses revealed that both SfABF51 isozymes can catalyze the versatile hydrolysis of α-(1,2)- and α-(1,3)-ʟ-arabinofuranosidic linkages of AXOS, and α-(1,2)-, α-(1,3)-, and α-(1,5)-linkages of linear and branched AOS. On the contrary, they have much lower activity against the α-(1,2)- and α-(1,3)-double-substituted substrates than the single-substituted ones. These hydrolases could potentially play important roles in the degradation and utilization of hemicellulosic biomass by S. fibuligera.

Examination of Genetic Relationships of Silkworm Stocks in Korea by Additive Isozyme Analysis

  • Sohn, Bong-Hee;Kim, Hyun-Su;Kang, Pil-Don;Lee, Sang-Uk;Seong, Su-Il
    • International Journal of Industrial Entomology and Biomaterials
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    • v.5 no.2
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    • pp.205-211
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    • 2002
  • Previously, genetic relationships among 321 silkworm races preserved in the Department of Sericulture and Entomology, NIAST, RDA were studied using six isozymes comprising 20 loci. As a part of additional studies, three additional isozymes with 7 loci [phos-phoglucomutase (PGM), glucose phosphate isomerase (GPI), malete dehydrogenase (MDH)] from hemolymphs, midgets, and eggs were employed to reevaluate the previous dendrogram (UPGMA method). Although the current study supported the recognition of the previously identified major groups, some minor changes were apparent among the selected 47 forms. The current study showed that the polymorphic loci of GPI from eggs and MDH from hemolymph appear to be responsible characters for generating a new major group in the genetic relationship. Interpreting the data with current additive isozymes may represent more robust genetic relatioships among 321 silkworm races preserved in Korea, until further evidence is available.

Ultrastructure and Activity Pattern of Peroxidase in Secretory Trichomes of Drosera capensis (장대끈끈이주걱 분비모의 미세구조와 peroxidase 활성)

  • Kim, Eun-Soo;Oh, Seung-Eun;Yu, Seong-Cheol
    • Applied Microscopy
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    • v.28 no.3
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    • pp.399-414
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    • 1998
  • Glandular trichomes present on the leaf surface of Drosera capensis were examined using transmission electron microscopy. A large number of stalked glands exist on the adaxial surfaces of the leaf blade. The secretory head is composed of two layers of secretory cells, one layer of middle cells, and the inner tracheids. The secretory cells contain rough endoplasmic reticulum, mitochondria, plastids, Golgi apparatus, and vacuoles. The secretory cells show prominent cell wall ingrowth, and thick cuticle restricted on the subcuticular wall. Frequently, the cuticle has some pores, canal-like structures, showing electron -dense granules being penetrated through them. Ultrastructural localization using diaminobenzidine showed the electron-dense deposits in the vacuole. No peroxidase activity was seen in the cell wall and cytolasm. The activity of peroxidase (POX) isozymes in Drosera which isoelectric point (pI) is 3.6 and some anionic POX isozymes which pIs are laid between 3.6 and 4.6 were especially increased according to the development and the formation of glandular trichomes. Also, the activity of some POX isozymes which isoelectric points are laid between 4.6 and 5.1 were increased in the regions of leaves which has trichomes.

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Genotypic and Geographical Variations of $\beta$amylase Isozyme in Soybean Land Races by Isoelectric Focusing (IEF)

  • Yoon, Mun-Sup;Ahn, Jong-Woong;Kang, Jung-Hoon;Baek, Hyung-Jin;Park, Nam-Kyu;Rho, Yong-Deok
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.2
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    • pp.139-142
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    • 2000
  • The experiment was carried out to study the variations and geographical distribution of $\beta$-amylase isozyme by isoelectric focusing (IEF) within Korean, Chinese and Japanese soybean land races. In pH 3-10 gel of IEF, the amylase of soybean accessions was separated into low pI group isozymes (TEX>${$Sp_1$}^b$) and high pI group isozymes(${$Sp_1$}^a$). In pH4-6.5 gel, isoelectric points were at 5.07, 5.15, 5.25, 5.40, and 5.94, and h, j, and k bands also were found. The distribution of Sp$_1$$^{a}$ allele (high pI type) was 29.3% in soybean accessions from Korea, 10.1 % in those from China, and 6.9% in Japanese accessions. The percentage of ${$Sp_1$}^a$) allele was the highest in soybean accessions from Kyungsang province (35 %) in Korea, then central China (32 %) in China, and Honshu (10%) in Japan.

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Effects of Intravenous Administration of Taurocholic Acid on Liver Lysosomal $\alpha$-D- and $\beta$D-Mannosidase Activities in Rats with Extrahepatic Cholestasis

  • Park, So-Kyung;Kim, You-Hee;Kwak, Chun-Sik
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.93-98
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    • 2004
  • The effects of intravenously administered of high concentration of taurocholic acid (TCA) on $\alpha$-D- and $\beta$-D-mannosidase activities in rat liver lysosomes were studied. These liver lysosomal enzymes, and serum lysosomal $\alpha$-D- and $\beta$-D-mannosidase isozymes activities were determined in experimental rats with common bile duct ligation (CBDL). The liver lysosomal $\beta$-D-mannosidase activity as well as the serum lysosomal $\alpha$-D- and $\beta$-D-mannosidase isozymes activities were found to be significantly increased in the CBDL plus TCA injected group than in the control group such as CBDL alone group. However, the liver lysosomal $\alpha$-D-mannosidase activity was found to be significantly decreased in the CBDL plus TCA injected group. The above results suggest that TCA repress the biosynthesis of the lysosomal $\alpha$-D-mannosidase and induce the biosynthesis of the lysosomal $\beta$-D-mannosidase in the liver. And that the elevated serum lysosomal $\alpha$-D- and $\beta$-D-mannosidase isozymes activities are most likely due to increased hepatocyte membrane permeability caused by TCA mediated liver cell necrosis.

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Studies on the Identification of Turfgrass by Electrophoresis (SDS-PAGE, PAGIF) (전기영동법 (SDS-PAGE, PAGIF)에 의한 잔디 분류에 관한 연구)

  • 박재복;김영후;이수영
    • Asian Journal of Turfgrass Science
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    • v.5 no.1
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    • pp.11-22
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    • 1991
  • This experiment was executed to investigate the possibility of the application of taxonomic method through the isoelectric focusing with polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoresis with seeds in the identification of turfgrasses. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to investigate the pattern of seed proteins which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The isoelectric focusing with polyacrylarnide gel was used to investigate the activity of the three isozymes of esterase, peroxidase and phosphoglucose isomerase which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The results were summarized as follows. 1. The difference of the patterns of seed proteins was observed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identification of intra-genus was easily detected. 2. The three isozymes of esterase, peroxidase and phosphoglucose isomerase were investigated through isoelectric focusing with polyacrylamide gel. As a result, esterase was most effective among three isozymes in the identification of turfgrass cultivars 3. In the past cultivar identification was primarily based on visual morphological characters, but there was a lot of difficulty. If we should use electrophoresis, we will be able to identifvturfgrass cultivars more effectively.

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Activity and Isozyme Profile of Antioxidative Enzymes at Booting Stage of Rice Treated with Cold Water

  • Kim Ki-Young;Kim Bo-Kyeong;Shin Mun-Sik;Choung Jin-Il;Ko Jae-Kweon;Kim Jung-Kon;Lim Jung-Hyun;Yun Song-Joon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.289-294
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    • 2004
  • This study was carried out to investigate the antioxidative enzymes and isozymes between chilling-tolerant and -susceptible varieties at the booting stage under cold water stress $(13^{\circ}C)$ in japonica rice. Total SOD, CAT, POX, and GR activities on the basis of protein were found to be important factors to defend cold water stress. Especially, SOD and CAT activities showed distinctive differences between chilling-tolerant and -susceptible varieties. Chilling-tolerant varieties were higher than chilling-susceptible varieties for SOD and CAT activities. One of eight isozyme bands for SOD was a inducible isoform. Three isozymes for CAT and one isozyme for POX were closely correlated with defense to cold water stress. Total GR activities except Stejaree 45 on the basis fresh weight and POX were increased by cold water stress, but there was no difference between chilling-tolerant and -sus­ceptible varieties.