• Title/Summary/Keyword: 동위효소

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칠성장어 (Lampetra japonica)젖 산수소이 탈효소 동위효소의 정제 및 면역화학

  • Jo, Seong-Gyu;Park, Seon-Yeong;Yeom, Jeong-Ju
    • The Korean Journal of Zoology
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    • v.36 no.4
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    • pp.505-513
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    • 1993
  • 칠성장어(Lampetro juponico) 골격근의 젖산수소이탈효소(EC 1. 1. 1.27, Lactate dehvdrogenase LDH) 동위효소 꺽지(Coreoperco herzi)의 LDH Aa 동위효소 및 소(Bos taurus)의 LDH B4 동위효소를 affinity chromatography로 정제하였고, 대구(Gadus macrocephcfus)의 LDH Ca 동위효소를 affinity chromatography와 OEAE-Sephacel chromatography로 정제하였다. 칠성장어 골격근 LDH 동위효소는 affinity chromatography에서 buffer를 유입한 후 용출되는 분획에서 모두 확인되었고, 정제한 칠성장어 골격근의 LDH에 대한 항체는 꺽지 LDH A4, C4 및 54 동위효소, 대구 LDH A4 및 B4 동위효소 그리고 생쥐(MUS musculus) LDH A4 등위효소와 복합체를 형성하였다. 칠성장어 골격근조직에서는 비각우 및 Ldh절가 발현되고 하부단위체 A의 구조는 하부단위체 B의 구조와 유사하며, 하부단위체 A는 진화상 보존적 이지만 하부단위체 C는 진화속도가 상당히 빠른 것으로 사료된다.

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Heterogeneity of Lactate Dehyrogenase Isozymes in tissues of Lampetra japonica (칠성장어(Lampetra japonica) 조직내 젖산수소이탈효소 동위효소들의 이질성)

  • 조성규;염정주
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.319-328
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    • 1993
  • All vertebrates other than lampreys exhibit multiple loci encoding lactate dehydrogenase (EC 1.1.1.27,LDH). From the result shown by cellulose acetate and starch gel electrophoresis, the lampreys were-reported to have only one isozyme. However in our results the LDH of skeletal muscle, heart and kidney in Lampetra japonica were separated into three isozymes and that of liver was separated into two isozymes by polyacrylamide gel electrophoresis. The LDH of skeletal muscle and heart were separated into four isozymes and that of liver was separated into two isozymes by polyacrylamide gel isoelectric focusing (PAGlEF). The LDH of skeletal muscle were separated into four isozymes through the chromatofocusing. The molecular weight of LDH isozymes in skeletal muscle was approximately estimated to be 140,000 by Sephadex G-200 gel filtration. The LDH isozymes of skeletal muscle, heart and liver were inhibited by pyruvate to the nearly similar degree. And the degree of inhibition by pyruvate showed the value between LDH A$_4$and LDH B$_4$isozyme. And the LDH isozymes in heart, liver and skeletal muscle were thermostable. The results mentioned above indicate that the LDH isozyme in lamprey (Lampetra japonica) has not one isozyme but isozymes. And it is also found out that the two structures of their subunits are similar each others.

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CNS Durg-induced Redistribution of Lactate Dehydrogenase Isozymes in Mice (CNS drug에 의한 mouse 젖산수소이탈효소 동위효소의 재분포)

  • 염정주;김상엽
    • The Korean Journal of Zoology
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    • v.32 no.1
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    • pp.34-39
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    • 1989
  • Ouabain, strychnine sulfate, caffeine sodium benzoate and chlorpromazine hydrochloride were introduced intraperitoneally into male mice for 7, 14 and 21 days to induce the changes in the relative percentages of lactate dehydrogenase isozymes. The five isozymes in brain, heart and kidney tissues were electrophoresed on cellulose acetate strip and subjected to densitometry. Ouabain caused a drastic increase of B$_4$isozymes only in brain tissues. The two stimulants altered the relative percentages of $A_4$and B$_4$isozymes conspicuously in brain tissues, whereas virtually no redistributions of five isozymes were occurred by the depressant except B$_4$isozymes in brain and heart tissues. On the basis of these observations, it might be suggested that the changes in intracellular concentration of sodium and calcium ions are not the cause of the isozyme redistributions and that Organization of plasma membrane could be one of the factors involved in the tissue specificity of lactate dehydrogenase isozymes in vertebrates.

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Variation of Lactate Dehydrogenase Isozymes in Angelfish (Pterophyllum scalare) according to Acute Environmental Change (급격한 환경변화에 대한 angelfish (Pterophyllum scalare) 젖산탈수소효소 동위효소의 변화)

  • An, Chang-Su;Cho, Sung-Kyu;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.3
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    • pp.416-423
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    • 2010
  • In this study, the properties and gene expression of the lactate dehydrogenase (EC 1.1.1.27, LDH) isozyme were studied in angelfish (Pterophyllum scalare) - known for their adaptation to the low oxygen environment of the tropics - which were acclimated to acute temperature change ($27{\pm}0.5{\rightarrow}18{\pm}0.5^{\circ}C$) and dissolved oxygen (DO) change ($6{\pm}1{\rightarrow}18\;ppm$) for 2 hours. The properties of the LDH isozymes were confirmed in the native-polyacrylamide gel electrophoresis, Western blot analysis and enzyme activity measurement. Liver- and eye-specific Ldh-C gene were expressed in liver, eye and brain tissues. Through Western blot analysis, the LDH $A_4$ isozyme was shown to have a more cathodal mobility relative to the $B_4$ isozyme. In the liver tissue, the LDH $A_4$ isozyme increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $C_4$ isozymes increased with DO increment, while the $B_4$ isozyme decreased. In the eye tissue, the LDH $A_4$ and B4 isozymse increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $B_4$ isozymes increased with DO increment, but the $C_4$ isozyme and isozymes including the subunit C decreased. In the heart tissue, LDH activity increased with DO increment, as well as the LDH $B_4$ isozyme. In the brain tissue, the LDH $A_4$ and $B_4$ isozymes increased with temperature drop. The LDH $B_4$ isozyme increased with DO increment. Accordingly, since the liver- and eye-specific Ldh-C are influenced by changes in DO and the LDH $B_4$ and $C_4$ isozymes are relatively controlled in the liver and eye tissues, the $C_4$ isozyme can be considered to have a lactate oxidase function.

Esterase Isozymes of Beet Armyworm, Spodoptera exigua(H bner), with Development and Tissues (발육 및 조직에 따른 파밤나방(Spodoptera exigua (H bner)) 에스테라제 동위효소)

  • 강성영;김용균
    • Korean journal of applied entomology
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    • v.37 no.2
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    • pp.179-185
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    • 1998
  • The effect of physiological factors of beet armyworm, Spodoptera exigua (Hubner), on esterase variation was analyzed by comparing electrophoretic esterase isozymes. Each esterase isozyme was also characterized by substrate and inhibitor specificities. A total of 28 esterase isozymes were separated on 10% nondenaturing polycarylamide gel electrophoresis (PAGE). These isozymes were denoted from El to E28 according to cathodal migration distances. There was a variation in esterase isozymes among developmental stages. Larvae and pupae had more isozymes than did adults. Eggs had only eight isozymes. The isozymes of El and E2 were specific only in the first instar larvae. Esterases also showed variation according to different tissues. More kinds of esterase isozymes were found in epidermis and gut tissues than in hemolymph and fat body. Some isozymes were specific in epidermis (from El to E6), gut (E10, El 1, E25, E26, and E27), and hemolymph (E18). Among 10 naphthyl esters, a-naphthyl propionate was the most reactive substrate to the esterase isozymes. The isozymes were classified into cholinesterases (El0 and E24), arylesterases (E4, E9, E17, E19, E21, and E23), and carboxylesterases (the others) on the basis of inhibition by the esterase inhibitors-eserine, dichlorovos, moncrotophos, and paraoxon.

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Purification, Kinetics and Immunochemistry of Two Homotetrameric Lactate Dehydrogenase Isozymes in Pseudogobio esocinlus (Cypriniformes) (Pseudogobio esocinus (Cypriniformes) 젖산수소이탈효소 동질사량체들의 정제, 역학 및 면역화학)

  • 김명옥;염정주
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.420-428
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    • 1989
  • Pseudogobio esocinus의 심장, 신장 및 간 조직은 하부단위체 C를 함유하는 젖산수소이탈효소를 갖고 있음이 확인되었다. 하부단위체 A 및 B에 대한 유전자들의 조직 발현은 다른 포유동물의 것과 유사하였으며 분자량은 140,000 정도로 추정되었다. Oxamate gel을 사용한 chromatography결과 A4 동위효소는 NAD+보다는 column buffer에 의해 용출되었다. B4 동위효소는 CM-Sepharose column을 사용하여 부붙 정제되었다. B4 동위효소는 물론 A4 동위효소도 고농도의 Pyruvate에 의해 저해되었다. A4 동위효소의 affinity chromatography 상 행동과 Pyruvate 저해 정도로 보아 A4 등위효소는 B4 동위효소 두 역학적으로 유사하다고 사료된다. P. esainus A4 동위효소에 대한 항체는 mouse A4 등위효소와 반응하지만 동종의 B4 동위 효소와는 반응하지 않는 특성으로 보아 하부단위체 B는 진화과정에서 보존성이 낮은 것으로 사료된다. Three tissues of heart, kidney and liver of a primitive cvprinid Pseudogobio esocinus were found to have lactate dehydrogenase isozyme(5) containing subunit C. Tissue expressions of genes for subunits A and B were similar to those of mammalian species. Molecular weight of the isozymes were estimated to be 140,000 approximately. Affinity chromatography of the isozymes on the immobilized oxamate gel revealed that A4 isozyme was not elected in NAD+ but in column buffer. B4 isozune was isozpnatically purified by subjecting kidney extract to a CM-Sepharose column. Ae isozvme as well as B4 isozvme was inhibited by high concentrations of pyruvate. The affinity chromatographic behavior and susceptibility to pyruvate inhibition of the A4 isorpne suggest that A4 isozwne is similar to B4 isozyme kinetically. Antibodies against p. esocinus A4 isogyme reacted with mouse At isozyme but not with p. esocinus B4 isogyme, reflecting that subunit B is less conservative in its evolution.

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Subunit Interactions of Vertebrate Lactate Dehydrogenase: II. Molecular Hybridizationa

  • Yum, Jung-Joo;Kim, Sang-Yeop;Park, Sang-Yoon
    • The Korean Journal of Zoology
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    • v.24 no.4
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    • pp.201-215
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    • 1981
  • In vitro intra- and interspecific molecular hybridizations of lactate dehydrogenase isozymes from six species were performed by freezing and thawing the isozymes in sodium chloride, resulting each of the interspecific hybridization of Fluta alba $H_4 \\times$ Ophicephalus argus $M_4$ and that of O. argus $M_4 \\times$ Parasilurus asotus isozymes showed one hybrid isozyme considered to be an $M_2H_2$ isozyme. The results fit with the suggestion that the isozyme of lower vertebrates is a dimer of homodimer as far as te assemblies of subunit M and subunit H are concerned.

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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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Effect of Light and Cadmium on the Activity and Isozyme Pattern of Catalase from Ric(Oryza sativa L.) (빛과 카드뮴이 벼 catalase 활성과 동위효소 발현에 미치는 영향)

  • Kim, Yoon-Kyoung;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.287-292
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    • 2006
  • The effects of cadmium on the catalase activity and isozyme patterns under light and dark conditions of rice(Oryza sativa L. cv. Dongjin) seedlings were examined. Cadmium treatment resulted in the notable enhancement of $H_2O_2$ contents in the seedling roots and leaves under light and dark conditions. The catalase isozyme patterns in the roots were different from those in the leaves, showing tissue-specific expression of the enzyme. Moreover, the expression patterns of catalase isozymes in the green seedling roots were different from those in the etiolated seedling roots following cadmium treatment. The increase of total catalase activity was about 16 times at 1 mM cadmium and marked inductions of the isozyme CAT1 and CAT2 contributed to this increase in the green seedling roots. On the other hand, in the etiolated seedling roots, total catalase activity was lower than that of control at 0.5 and 1 mM cadmium, even though catalase activity increased about 3 times at 0.1 mM cadmium. The 3 fold increase of total catalase activity was mainly due to the increase of CAT1, CAT3 and CAT4 at 0.1 mM cadmium. However, treatment with higher concentrations of cadmium decreased the activity of CAT2 and CAT4 in the etiolated roots. In the leaves, the catalase existed as three isozymes; one cationic isozyme CATc, one neutral isozyme CATn and one anionic isozyme CAT1 in the control. The isozyme patterns and total activities remained unaffected by cadmium under light and dark conditions in the seedling leaves. Taken together, it seems that cadmium-induced changes of catalase might be regulated by light in the roots, but not in the leaves.

Effects of Different Exercise Intensities on Cytosolic and Mitochondrial LDH Isozymes of Cardiac Muscle in Rats (운동강도의 차이가 흰쥐의 심근 세포기질 및 미토콘드리아 LDH 동위효소 변화에 미치는 영향)

  • Lee Sang-Hak;Yoon Jin-Hwan
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.80-86
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    • 2005
  • To achieve the purpose of this study, forty-eight male Sprague-Dawley rats were assigned to control and three endurance exercise group. 36 rats were forced to exercise according to exercise intensity for 8 weeks and 12 rats were untrained for control group. Cardiac cytosol was extracted from cardiac tissue and cardiac mitochondria was purified from the cytosol. Purified mitochondria were separated into four fraction: inner membrane, outer membrane inter membrane space and matrix. The changes of cytosolic and mitochondrial LDH isozymes activity were measure. Relative activity $(\%)$ of cytosol for low and control group showed the following order of prevalence $AB_3>A_2B_2>B_4>A_3B>A_4$ for moderate and high group : $AB_3>B_4>A_2B_2>A_3B>A_4$. Outer membrane for low group showed $AB_3>B_4>A_2B_2$, for moderate group:$ B_4>AB_3>A_2B_2$, for high and control group: $B_4>A_3B$. Inter membrane space for low, moderate and high group showed $B_4>AB_3>A_2B_2>A_3B>A_4$, for control group: $B_4>A_3B>AB_3>A_2B_2>A_4$. Inner membrane for all group showed $B_4>AB_3>A_2B_2>A_3B>A_4$. Matrix for control, low, moderate and high group showed $B_4>AB_3>A_2B_2>A_3B>A_4$. These results suggest that long term exercise intensity effect on cardiac tissue cytosolic and mitochondrial activity and $A_4,\;A_3B,\;A_2B_2,\;AB_3\;and\;B_4$ isozymes were found entirely in mitochondrial fraction.