• Title/Summary/Keyword: 동위효소

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Forest Genetics of Isozymes (임목(林木)의 동위효소(同位酵素))

  • Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.43 no.1
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    • pp.74-86
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    • 1979
  • 동위효소(同位酵素)가 임목분야(林木分野)의 연구(硏究)에 이용(利用)되기 시작(始作)한지 10여년이 지난 요즈음 많은 분야(分野)에서 그 성과(成果)가 높이 평가(評價)되고 있다. 특(特)히 복잡(複雜)한 유전변이(遺傳變異)를 가지고 있는 천연임분(天然林分)에서의 유전분석(遺傳分析)으로 많은 것을 밝혀 내어 임목(林木)의 유전육종학적(遺傳育種學的) 측면(側面)뿐만 아니라 삼림유전생태학적(森林遺傳生態學的)인 면(面)에서도 큰 도움을 주고 있다. 본(本) 총설(總說)은 지금까지 임목(林木)에 대(對)한 동위효소(同位酵素) 이용(利用) 실례(實例)를 중심(中心)으로 앞으로 응용(應用) 가능분야(可能分野)에 대(對)해 검토(檢討)한 것이다.

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Studies on the Intraspecific Variations on Geological Distributions of Acusta despecta sieboldiana in Korea (한국산 달팽이 ( Acusta despecta sieboldiana) 의 지리적 분포에 따른 종내 변이에 관한 연구)

  • 이준상;권오길
    • The Korean Journal of Malacology
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    • v.12 no.1
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    • pp.19-31
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    • 1996
  • 국내 11개 지역(경기도 옹진군 덕적도, 경기도 의정부시 소요산, 강원도 춘천시, 충청남도 태안군 안홍, 경상북도 울릉군 울릉도, 전라북도 부안군 변산, 전라남도 신안군 홍도, 흑산도 비금도, 진도, 제주도)에서 채집된 달팽이 (A. despecta sieboleiana)를 대상으로 외부형태 분석과 동위효소를 검출하여 각 지역 집단간의 형태적, 유전적 유연관계와 변이 정도를 연구한 결과는 다음과 같다. 외부형태에 의한 집단간 유연관계는 안홍과 덕적도집단(average taxonomic distance, D=0.358)이 형태적으로 가장 유사한 집단으로 나타났다. Polyacrylamide gel을 사용하여 11가지의 동위효소를 검출한 결과 AKP, ACP, AO, EST, GPD, HBDH, LDH, SDH, XDH의 동위효소에서 유전적 다형현상이 나타났다. 전 집단의 평균 다형형의 빈도는 59.19%이며 이형접합자 빈도는 0.263이고 각 집단의 유전적 차이치는 춘천과 울릉도 집단이 0.066(genetic distance)으로 가장 높은 유사성을 보였다. 지리적 분포에 따른 외부형태 변이와 유전적 변이와의 관계는 유의성이 없었다.

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Characteristics of alkaline and acid phosphatase in Spirometra erinacei (만손열두조충(Spirometra erinacei)에서 알칼리성과 산성 인산효소의 특성)

  • 곽기훈;김창환
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.69-78
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    • 1996
  • This study was done to investigate the enzyme-histochemical localization and characteristics of alkaline and acid phosphatase related with metabolism in sparganum and adult of Spirometrn erinacei. By the enzyme-histochemical assay, the alkaline and acid phosphatases were localized in the tegument and subtegumental musculature of sparganum and adult, but not in the parenchyma. The activities of alkaline phosphatase were stronger in the tegument than in the subtegumental musculature, and activities of acid phosphatase were stronger in the tegument of adults than those of sparganum. The 2 isozymes of alkaline and acid phosphatases were separated from s-sparganum (from snake) and r-sparganum (from experimentaly infected rats) respectively but 4 isozymes of Alp and 3 isoxymes of Acp were separated from adult worms by electrophoresis. In isogyme Alp, the 661)a was the common isozyme, but 130 kDa isozyme of Acp was the common isozyme in spargana and adult worms. By isoelectrofocusing, 4 isozymes (PI 7.9, 7.7, 6.5 and 6.3) and 2 isozymes (PI 7.9 and 7.7) of alkaline phosphatase were separated from adults and spargana respectively. In the stability against heat, activity of alkaline phosphatase was denatured perfectly after heating at 90℃ for 40 seconds. The optimum pH and temperature for activity of alkaline phosphatase were about pH 10 and 50℃, respectively. The maximum activity (unit) of alkaline phosphatase was 22.0 in s- sparganum,25.0 in r-sparganum and 215.0 in adult worms, so that the maximum activity was revealed higher in adults than spargana. As the result from above, we observed that alkaline and acid phosphatases were functioned mainly in the tegument and subtegumental musculature , and the isoxymes of phosphatase were activated differently according to habitat of the parasites. The spargana and adult worms carry out the pafasitism by adapting thenlselves to parasitic circumstance loth these emxymes.

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Serum Enzyme and Isozyme Activities of Rats Acclimated to Cold Environment (寒冷環境에 순화시킨 흰쥐의 血淸酵素 및 同位酵素의 活性)

  • 정애순;남상열
    • The Korean Journal of Zoology
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    • v.29 no.2
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    • pp.107-120
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    • 1986
  • The activities of serum of serum lactate dehydrogenase (SLDH), serum alkaline phosphatase (SALP), serum creatine phosphokinase (SCPK), and their isozymes were determined in adult male Sprague-Dawley rats acclimated to cold environment $(4\\pm1^\\circC)$ for 36 days. The SLDH activity was significantly higher in the early stage of acclimated period. The steady state of SLDH activity seemed to be reached by the end of acclimated period. Electrophoretic separation of serum of control rat showed three SLDH isozymes. Isozymes SLDH4 and SLDH5 appeared most prominently, whereas only trace of SLDH1 or SLDH2 was found. The increase in SLDH level during acclimation to cold environment is a reflection of an immediate increase in the SLDH1, SLDH2, and SLDH3 type of SLDH isozyme. The acclimation to cold environment increased significantly level of SALP in the early state of acclimated period. SALP activity showed a attaining steady state with the resting level after transient rise. Electrophoretic separation of SALP of control rats showed the SALP1 and SALP2 fractions. The transient rise in SALP activity of rats acclimated to cold environment coincided with a transient rise in SALP1 fraction. Immediately after exposure to cold environment, there was significant elevation in SCPK activity. Value returned to normal after transient rise. A new steady state of SCPK activity seemed to be reached by 36 days. It may be inferred from the above data that thermal compensation appears to result from a change in the activity of an enzyme and that the SLDH, SLDH-isozyme, SALP-isozyme, and SCPK may be involved directly or indirectly in thermoregulation during acclimation to cold environment.

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Characterization of Lactate Dehydrogenase in Acanthogobius hasta (풀망둑(Acanthogobius hasta) 젖산탈수소효소의 특성)

  • Yum, Jung-Joo
    • Journal of Life Science
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    • v.18 no.2
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    • pp.264-272
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    • 2008
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.

Vegetative Compatibility, Isozyme Polymorphisms and Pathogenicity of Isolates of Fusarium oxysporum f. sp. fragariae (딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 균주간 Vegetative Compatibility, 동위효소 Polymorphism과 병원성)

  • 현재욱;김순옥;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.33-40
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    • 1996
  • 우리나라 여러 딸기 재배지에서 전형적인 시들음 증상을 나타내는 이병식물에서 분리한 32개의 Fusarium oxysporum 균주들을 vegetative compatibility와 전기영동에 의한 동위효소상의 차이에 의해 분류하고 이들과 병원성과의 관계를 알아보았다. Nitrate nonutilizing(nit) mutant를 이용하여 vegetative compatibility group으로 분류해 본 결과 크게 A, B, C, D의 4개 그룹으로 나눌 수 있었으며 A그룹에는 15개 균주, B그룹에는 7개 균주, C와 D그룹에는 각각 2개 균주, 그리고 single VCG인 6개 균주가 존재하였다. 전기영동에 의한 esterase, catalase, acid phosphatase, leucin-aminopeptidase(LAP)의 동위효소상을 비교하여 본 결과 I, II, III, IV의 4개 그룹으로 나눌 수 있었으 I 그룹에는 18개 균주, II그룹에는 2개 균주, III그룹에는 6개 균주, IV그룹에는 6개 균주로 분류할 수 있었다. 또한, VCG의 A와 D그룹에 속하는 모든 균주들의 동위효소상의 I 그룹에 속하였으며 VCG B그룹의 7개 균주들 중에서 5개 균주가 동위효소상의 IV그룹, 7개 균주들 중에서 5개 균주가 동위효소상의 IV그룹, 나머지 2개 균주는 I 그룹과 III 그룹에 속하였으며 C그룹의 균주는 III과 IV 그룹에 속하였고, single VCGs들은 III 그룹에 4개 균주, II 그룹에 2개 균주가 속하므로 VCG와 동위효소상 간에는 밀접한 관계가 있음을 알 수 있었다. 4개 딸기 품종에 대해 병원성을 비교하여 본 결과 VCG A에서 선발된 2개 균주들은 보교조생에 대해 가장 병원성이 강하였으며 정보나 여홍에 대해서도 강한 병원성을 가지고 있었다. 반면에 B 그룹에서 선발된 2개 균주들은 4개 품종에 대해 병원성이 약하거나 거의 없었으며 C 그룹에서 선발된 1개 균주는 여홍에 대해 상대적으로 가장 강한 병원성을 가지고 있었다.

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Effect of $GA_3$ and ABA on Peroxidase and Catalase Activities and Isoperoxidase Patterns in Mung Bean Seedling (녹두의 발아과정 중 $GA_3$ 및 ABA의 처리가 Peroxidase, Catalase 활성변화와 Isoperoxidase Pattern에 미치는 영향)

  • Lee, Sang-Kap;Park, Woo-Churl
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.205-210
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    • 1988
  • The changes in peroxidase and catalase activities, and isoperoxidase patterns in different parts of mung bean seedling caused by the treatment with plant growth substances, $GA_3$ and ABA, were examined. As germination proceeded, the activity of peroxidase in all part except hypocotyl was increased, while that of catalase decreased. The separate application of $GA_3$ and ABA increased the activity of peroxidase which was more influenced by $GA_3$ than by ABA only in cotyledon, while that of catalase was more affected by ABA than by $GA_3$. Electrophoretic study revealed that the number of isoperoxidase was increased continuously in all parts during development. A greater influence was exerted on the intensity of isozyme than the number of isozyme by the hormonal treatment.

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Expression of Phospholipase C Isozymes in Human Lung Cancer Tissues (인체 폐암조직에서 Phospholipase C 동위효소의 발현양상)

  • Hwang, Sung-Chul;Mah, Kyung-Ae;Choi, So-Yeon;Oh, Yoon-Jung;Choi, Young-In;Kim, Deog-Ki;Lee, Hyung-Noh;Choi, Young-Hwa;Park, Kwang-Ju;Lee, Yi-Hyeong;Lee, Kyi-Beom;Ha, Mahn-Joon;Bae, Yoon-Su
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.310-322
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    • 2000
  • Background : Phospholipase C(PLC) plays an important role in cellular signal transduction and is thought to be critical in cellular growth, differentiation and transformation of certain malignancies. Two second messengers produced from the enzymatic action of PLC are diacylglycerol (DAG) and inositol 1, 4, 5-trisphosphate (IP3). These two second messengers are important in down stream signal activation of protein kinase C and intracellular calcium elevation. In addition, functional domains of the PLC isozymes, such as Src homology 2 (SH2) domain, Src homology 3 (SH3) domain, and pleckstrin homology (PH) domain play crucial roles in protein translocation, lipid membrane modificailon and intracellular memrane trafficking which occur during various mitogenic processes. We have previously reported the presence of PLC-${\gamma}1$, ${\gamma}2$, ${\beta}1$, ${\beta}3$, and ${\delta}1$ isozymes in normal human lung tissue and tyrosine-kinase-independent activation of phospholipase C-${\gamma}$ isozymes by tau protein and AHNAK. We had also found that the expression of AHNAK protein was markedly increased in various mstologic types of lung can∞r tissues as compared to the normallungs. However, the report concerning expression of various PLC isozymes in lung canærs and other lung diseases is lacking. Therefore, in this study we examined the expression of PLC isozymes in the paired surgical specimens taken from lung cancer patients. Methods : Surgically resected lung cancer tissue samples taken from thirty seven patients and their paired normal control lungs from the same patients, The expression of various PLC isozymes were studied. Western blot analysis of the tissue extracts for the PLC isozymes and immunohistochemistry was performed on typical samples for localization of the isozyme. Results : In 16 of 18 squamous cell carcinomas, the expression of PLC-${\gamma}1$ was increased. PLC-${\gamma}1$ was also found to be increased in all of 15 adenocarcinoma patients. In most of the non-small cell lung cancer tissues we had examined, expression of PLC-${\delta}1$ was decreased. However, the expression of PLC-${\delta}1$ was markedly increased in 3 adenocarcinomas and 3 squamous carcinomas. Although the numbers were small, in all 4 cases of small cell lung cancer tissues, the expression of PLC-${\delta}1$ was nearly absent. Conclusion : We found increased expression of PLC-${\gamma}1$ isozyme in lung cancer tissues. Results of this study, taken together with our earlier findings of AHNAK protein-a putative PLD-${\gamma}$, activator-over-expression, and the changes observed in PLC-${\delta}1$ in primary human lung cancers may provide a possible insight into the derranged calcium-inositol signaling pathways leading to the lung malignancies.

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Lactate Dehydrogenase and Monocarboxylate Transporters 1, 2, and 4 in Tissues of Micropterus salmoides (큰입우럭(Micropterus salmoides) 조직의 젖산탈수소효소 및 Monocarboxylate 수송체(MCT) 1, 2, 4)

  • Yum, Jung-Joo;Yeon, Jun-Hee
    • Journal of Life Science
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    • v.22 no.1
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    • pp.98-109
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    • 2012
  • The properties of lactate dehydrogenase (EC 1.1.1.27, LDH) and expression of monocarboxylate transporters (MCTs) 1, 2, and 4 were studied in tissues from Micropterus salmoides. Native-PAGE revealed that the LDH $A_4$ isozyme was predominantly located in skeletal muscle. The LDH $A_4$, $A_2B_2$, and $B_4$ isozymes were detected in heart, liver, eye, and brain tissues, while eye-specific $C_4$ isozyme was detected in eye tissue. In September, strong LDH $B_4$ isozyme activity was detected in heart tissue. High $A_4$ isozyme activity was noted in all other tissues except heart tissue. However, in November, strong $A_4$ isozyme activity was detected in heart tissue. The LDH/CS (Citrate synthase, EC 4.1.3.7) ratio in skeletal muscle and heart tissues indicated that anaerobic metabolism was high in those tissues. Native-PAGE after immunoprecipitation showed that eye-specific $C_4$ isozyme was more similar to the $A_4$ than the $B_4$ isozyme. The LDH $A_4$ isozyme was purified by affinity chromatography. The molecular weight of subunit A was 37,200. The LDH activity in tissues was consistently 11.05~28.32% due to inhibition by 10 mM pyruvate. The $K_m^{PYR}$ of LDH in eye tissue was very low. The optimum pH for LDH in tissues was pH 7.5~8.0. The LDH $A_4$ isozyme was detected in mitochondria of skeletal muscle, whereas the $B_4$ and $A_2B_2$ isozymes were detected in heart tissue mitochondria. Western blot analysis indicated that MCTs 1, 2, and 4 were located in the plasma membrane and mitochondria of skeletal muscle and heart tissues. The sizes of MCTs 1, 2, and 4 in skeletal muscle were 60, 54~38, and 63 kDa, while those in heart tissue were 57, 54~38, and 55.5 kDa, respectively. In conclusion, M. salmoides appears to use anaerobic metabolism predominantly when adapted to a hypoxic environment. In highly activated skeletal muscle and heart tissue, energy production is controlled by inward and outward flows of pyruvate and lactate through MCTs 1, 2, and 4 in the plasma membrane and mitochondria, with effective adjustment by LDH isozymes.

Lactate Dehydrogenase Isozyme of Hypoxia Tropical Catfish(Pangasius Polyuranodon, Hypostomus Plecostomus) (저산소 환경에 서식하는 열대성 catfish (Pangasius Polyuranodon, Hypostomus Ple-Costomus)의 젖산탈수소효소 동위효소)

  • 조성규;염정주
    • Journal of Life Science
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    • v.14 no.4
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    • pp.702-707
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    • 2004
  • In native-polyacrylamide gel electrophoresis of Pangasius polyuranodon, the lactate dehydrogenase (EC 1.1.1.27, LDH) $A_4$, $A_3$B, $A_2$$B_2$,$AB_3$ and $B_4$ isozymes were expressed in various tissues. The LDH $A_4$ and liver-specific $C_4$ isozymes were expressed in the tissues of Hypostomus Plecostomus. The bands of LDH in skeletal muscle, heart and eye tissues were not detected while one band was detected in kidney and liver, and four bands were detected in brain. The detected one band in liver was identified as alcohol dehydrogenase and an anodal band of skeletal muscle was identified as nothing dehydrogenase. The LDH in skeletal muscle, heart and eye might function as pyruvate reductase. The degree of inhibitions of LDH in skeletal muscle and heart of P. polyuranodon by 10 mM pyruvate were measured 57.6% and 73.8%, respectively. However, those of LDH in tissues of H. plecostomus were measured 52.7-61.8% so tissue specificity did not appear. Therefore, H. ple-costomus might be more acclimated to hypoxic environment by anaerobic metabolism of LDH iso-zymes than P. polyuranodon.