• Title/Summary/Keyword: $A_4$ isozyme

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Characterization and Evolutionary Relationship of Lactate Dehydrogenase in Liver of Lampetra japonica and Liver-specific C4 Isozyme in Gadus macrocephdus. (칠성장어(Lampetra japnica) 간조직 젖산탈수소효소와 대구(Gadus macrocephalus) liver-Specific C4동위효소의 특성 및 진화적 관계)

  • 박선영;조성규;염정주
    • Journal of Life Science
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    • v.14 no.4
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    • pp.708-715
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    • 2004
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) in liver of Lempetra japonica was purified in buffer of affinity chromatography. The liver-specific $C_4$ isozyme of Gadus macrocephalus was purified by heat treatment, affinity chromatography, and DEAE-Sephacel chromatography. The liver-specific $C_4$ isozyme was eluted in a buffer containing NAD+ and was coeluted with $B_4$isozyme in plain buffer of affinity chromagraphy. Liver-specific $C_4$ isozyme in G. macrocephalus was the most thermostable, and$B_4$isozyme was more stable than $A_4$. The LDH in the fraction of pH 7.45 purified from the liver of L. iaponica by chromatofocusing was more inhibited by pyruvate than purified LDH. The optimum pH of the LDH isozyme in the liver of L. japonica was 7.5 and that of liver-specific$C_4$ isozyme was 8.5. The LDH in liver of L. japonica made complexes more with antibody against Coreoperca herzi$A_4$ and liver-specific $C_4$ than with that against eye-specific $C_4$. Therefore, the structure of the LDH in liver of L. japonica might be similarly evolved to that of subunit A and liver-specific $C_4$ isozyme in liver tissue of G. macrocephalus. The evolution rate of subunit C is faster than that of subunit A. LDH in liver of L. japonica has not one isozyme but isozymes and it was also found out to have not only subunit A and B but also subunit C.

Purification and Characterization of Peroxidase Isozyme C from Mung Bean (녹두의 Peroxidase Isozyme C의 생화학적 성장)

  • Lee, Sang-Kap;Park, Woo-Churl
    • Applied Biological Chemistry
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    • v.30 no.3
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    • pp.219-226
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    • 1987
  • Peroxidase isozyme C was isolated from mung bean cotyledon and purified to homogeneity as ascertained by chromatography and polyacrylamide gel electrophoresis, and then crystallized. Purification procedures included ammonium sulfate precipitation and column chromatography on Sephadex G-75, DEAE-cellulose and DEAE-Sephadex A-50. Peroxidase isozyme C was purified about 63 fold with 5% recovery. Isozyme C showed optimal activity at pH 5.0 with o-dianisidine and at pH 6.0 with guaiacol as substrate, and the optimal temperature was $70^{\circ}C$. Molecular weight of 50,000 was estimated for the isozyme C by SDS-polyacrylamide gel electrophoresis. At $70^{\circ}C$, it took 30 min to inactivate the isozyme to 50%, and at $80^{\circ}C$, this isozyme was almost completely inactivated in 20 min. The Km value of isozyme C for o-dianisidine was 0.11mM and that for guaiacol was 60.98mM using hydrogen peroxide as cosubstrate, and the kinetic pattern showed a competitive cyanide inhibition with respect to substrate. The crystalline structure of isozyme C was rectangular in shape.

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척추동물의 Isozyme에 관한 비교연구: IV. 한국산 개구리목의 Lactate 및 Malate Dehydrogenase Isozyme

  • 고정식;조동현;박상윤
    • The Korean Journal of Zoology
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    • v.16 no.3
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    • pp.193-201
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    • 1973
  • A cellulose acetate electrophoretic survey of Korean Anura has revealed the presence of diverse lactate and malate dehydrogenase isozymes. The LDH and MDH isozymes in the tissues of the brain, heart, stomach, skeletal muscle and liver of the six species of Anura examined show the species specific patterns which differ from those of mammals and birds. Two isozymic forms of LDH and MDH exist in both Rana nigromaculata and Rana nigromaculata coreana, respectively, with almost the same pattern. LDH of Bombina orientalis has five isozymic forms, and that of Rana temporaria ornativentris contains four isozymes. Bufo sp. has 3 to 5, and Rana rugosa has 3 to 4 isozymic bands according to the tissues. MDH's of all animals have two isozymic forms with different spacing on the zymograms.

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Genetic Divergence and Relationship Among Four Abalone Species by Isozyme and AFLP analyses (Isozyme 및 AFLP분석에 의한 전복류 4종간의 유전적 차이 및 유연관계)

  • Park Choul-ji;Kijima Akihiro
    • Journal of Aquaculture
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    • v.18 no.4
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    • pp.252-259
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    • 2005
  • Isozyme and AFLP analyses were examined to estimate the utilities of them as a genetic marker. The utilities were evaluated by genetic divergence and relationships among the four distinct abalone species; Haliotis discus hannai collected from northeast coast of Japan and Yellow-Sea coast of China, H. rufescens collected from west coast of USA, H rubra collected from southeast coast of Australia and H midae collected from Cape Town of South Africa. Isozyme and AFLP analyses showed a clear genetic divergence between every pair of species. Genetic relationships among the low species estimated by isozyme and AFLP analyses reflected to geographical distribution and morphological characteristics. In conclusion, Isozyme and AFLP analyses are suitable genetic markers far estimates of genetic divergence and relationship among abalone species.

A Comparison of the Lactate Dehydrogenase (LDH) Isozyme Patterns in Vertebrate Cerebrum and Retina (脊椎動物 腦와 網膜에 있어서 Lactate Dehydrogenase Isozyme Pattern의 比較)

  • Kim, Soon Ok;Park, Sang Yoon
    • The Korean Journal of Zoology
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    • v.22 no.1
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    • pp.1-10
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    • 1979
  • From the experimental results of cellulose acetate membrane electrophoresis we concluded the followings in explaining the LDH isozyme patterns found in the retina and cerebrum of vertebrata. Lactate dehydrogenase of the retina and cerebrum of both Carassinus carassinus and Cyprinus carpio was found to have one diffused band located between $LDH_2$ and $LDH_1$. LDH isozyme patterns of heart, pectoral muscle, liver and stomach of the Cyprinus carpio had the same diffused band in all organs. LDH isozyme patterns of the cerebrum of Hynobius leechii and Rana nigromaculata were observed to be different, in Hynobius leeichi a single band moved to the negative pole and two bands of $LDH_5$ and $LDH_4$ were obtained in the Rana nigromaculata. The retina and cerebrum of Natrix tigrina lateralis were observed as one band but amyda maakii had different LDH isozymes of the retina and cerebrum. The retina of Amyda maakii had five distinct LDH isozyme bands which had decreasing activity in the order of $LDH_5, LDH_4, LDH_3, LDH_2 and LDH_1$. The cerebrum of Amyda maakii had one band like Natrix tigrina lateralis but it moved to the negative pole. LDH isozymes in the retina and cerebrum of Gallus gallus domesticus and Melopsittacus undulatus showed one band. Five characteristic LDH isozyme bands were obtained from the retina of mammals, Oryctolagus cuniclus, Canis familiaris, Sus scrofa bos taurus and in the cerebrum of mouse, albino rat, Rhinolophus ferrum-equinum kokai.

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Esterase Isozyme Variation in the Tissues of Glycine soja S. and Z. , Vigna vexillata var. tsusimensis Matsmura , and Trifolium repens L. in Korea (돌콩 , 돌동부 , 화이트 클로버의 조직별 Esterase Isozyme 변이에 관한 연구)

  • 이성규
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.12 no.4
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    • pp.253-259
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    • 1992
  • The esterase isozyme in tissue of wild legume plants were separated by horizontal starch gel electrophoresis. Extracts used in this study were prepared from fully expanded young leaf, cotyledon and radicle of seedling and root-nodule of Glycine sola, Vigna vexillata var. tsuscmensis and Trifoliwn repens. The results are as follows; 1. Each tissue examined had a characteristic banding pattcrn. Number of bands in each species, G. soja, V. vexillata, and T . repens, were 14, 8 and 1 1 bands, respectively. And difference in esterase isozyme bands were greater from tissue to tissue than difference between habitat. 2. Est-I, Est-2. Est-3 and Est-4 in G. soja, Est-I in V. vexillata and Est-l and Est-2 in T. repens showed strong cnzyme activity than other enzyme. 3. Esterase isozyme variation in G. soja and T . repens showed more variety than V. vexillata. This is resulted from many genotypic differences within species. 4. The main enLyme among thc esterase isozyme were Est-I. Est-2, Est-3 and Est-4.

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Electrophoresis Techniques for Identification of Callus Induced from Pinella ternata (Thunb.) Breit 1. Analysis of protein and Enzymes of Callus Induced from Pinella ternata (Thunb.) Breit (기내 배양된 반하의 전기영동에 의한 생약학적 연구 1. 전기영동법에 의한 반하 Callus의 단백질 및 효소분석)

  • Choi, Jung-Sik;Ryu, Jeom-Ho;Park, Hak-Bong;Kim, Hyung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.33 no.2
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    • pp.122-125
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    • 1988
  • A comparative electrophoretic study on protein and several important enzymes of calli induced from stem, intercostal area and minor vein area were conducted in Pinella ternata (Thunb.) Breit. Soluble protein band patterns of calli induced from stem, intercostal area and minor vein area were distinctly different from those of the corresponding plant parts. Esterase isozyme patterns of calli induced from stem, intercostal area and minor vein area were different from those of the corresponding plant parts. Glutamate oxalo-acetate transaminase isozyme patterns of calli grown for 4 weeks induced from 3 plant parts were similar to those of the corresponding plant parts. But a high molecular weight isozyme band appeared in the calli grown for 8 weeks. Alow molecular weight isozyme band disappeared on the peroxidase isozyme patterns of calli grown for 4 weeks appeared on those of the corresponding plant parts.

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Purification and Characterization of Eye-Specific Lactate Dehydrogenase C4 Isozyme in Greenling (Hexagrammos otakii) (쥐노래미 eye-specific LDH C4 동위효소의 정제 및 특성)

  • Cho, Sung-Kyu;Yum, Jung-Joo
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1565-1572
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    • 2011
  • Eye-specific lactate dehydrogenase (EC 1.1.1.27, LDH) $C_4$ isozyme in the eyes of greenlings (Hexagrammos otakii) was successfully purified by affinity chromatography and continuous-elution electrophoresis. The molecular weight of the purified eye-specific LDH $C_4$ isozyme was 154.8 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Optimal pH for enzymatic reaction of the eye-specific LDH $C_4$ isozyme was pH 8.5. $K^{PYR}_m$ value of the purified eye-specific LDH $C_4$ isozyme was $1.88{\times}10^{-5}$ M using pyruvate as a substrate. These results indicate that we must consider pH when measuring eye-specific LDH $C_4$ isozyme activity. The eye-specific LDH $C_4$ isozyme had a higher binding affinity for the substrate as a pyruvate than LDH A4 isozyme. Antibodies produced against the purified eye-specific LDH $C_4$ isozyme may be used in the diagnosis of several human diseases and in comparative physiological studies of fishes.

Enzyme-Htstochemical Studie5 of Esterase and Phosphatase on Developing Spirometra erinacei (Spirometra ernacei의 발육에 따른 Esterase와 Phosphatase의 조직 화학적 연구)

  • 곽기훈;김창환
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.225-235
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    • 1988
  • This study was carried out to compare distribution and isozyme pattern of nonspecific esterase, acid phosphatase and alkaline phosphatase on developing sparganum and adult of Spinometra erinacei by using enzyme-histochemical method and electrophoresis The sparganum and adult were recovered from rats and cat that were infected by sparganum. The results obtained were as follows: Nonspecific esterase had a strong activity in the parenchymal musculature of sparganum and adult, but no detectable level in ihe tegument. A total of 7 and 8 nonspecific esterase bands were detectable in sparganum and adult, respectively. Of these bands, band 3 and 4 were major bands in sparganum and adult. Acid phosphatase had a strong activity in the tegument and the epidermal musculature of sparganum, but no detectable level in the parenchymal musculature. A total of 3 bands were detectable in sparganum and adult. Of these bands, band 3 was major band in sparganum and adult. Alkaline phosphatase had a strong activity in the tegument and the epidennal musculature of sparganum and of adult, but no detectable level in the parenchymal musculature. A total of 2 and 4 bands were detectable in sparganum and adult. Of these bands, band 2 was major band in sparganum and adult. Based on the present results isozyme band patterns showed qualitative and quantitative changes in each tissues of sparganum and of adult during the development.

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Changes of Activities and Isozymes of Lactate Dehydrogenase in Coreoperca herzi and Pseudogobio esocinus Acclimated to Rapid Increase of Dissolved Oxygen (급격한 용존산소량 증가에 순응한 꺽지(Coreoperca herzi)와 모래무지(Pseudogobioesocinus) 젖산탈수소효소 활성과 동위효소의 변화)

  • Cho Sung Kyu;Yum Jung Joo
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.71-79
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    • 2005
  • The metabolism of lactate dehydrogenase (EC 1.1.1.27, LDH) and $C_4$ isozyme were studied in tissues of Coreoperca herzi and Pseudogobio esocinus acclimated to rapid increase of dissolved oxygen (DO). In C. herzi the LDH activity was changed $35-39\%$ in brain and liver tissues, and within $20\%$ in other tissues. The $B_4$ isozyme was increased and isozyme containing subunit C was decreased in muscle tissue. The $B_4$ isozyme was increased in heart and kidney. In P. esocinus, the LDH activity in liver tissues was largely increased to $150\%$ for 30 minute and $70\%$ in other tissues. The $A_4$ isozyme was increased in muscle and $B_4$ isozyme was increased in other tissues. Especially, the metabolism of liver tissue in P. esocinus was regulated by increasing liver-specific $C_4$ and decreasing $A_4$ isozyme. But the metabolism of eye tissue in C. herzi was regulated by decreasing LDH activity and eye-specific $C_4$ isozyme. The LDH activity and LDH isozyme in P. esocinus were largely increased than C. herzi acclimated to rapid increase of DO. And eye-specific $C_4$ and liver-specific $C_4$ isozymes played role as lactate oxidase. Therefore, the response of species acclimated to rapid increase of DO seems to be variable, perhaps due to prior exposure to environmental conditions.