• The Korean Journal of Zoology
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• v.24 no.4
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• pp.173-188
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• 1981

근세포의 분화에 있어서의 근특이 단백질의 합성 순서를 구명하기 위하여 계배 근세포를 2$\\sim$9일간 배양하면서 단백질합성야상을 SDS-polyacrylamide 겔전기 영동법, 등전점초점2차원 전기영동법 및 방사자기법으로 분석하였다. Actin은 분화의 초기부터 활발히 합성되어 그 양이 다량으로 축적되나, myosin은 배양 3일째부터 대량 합성되기 시작하였다. Myosin의 대량합성시기는 배양 근원세포가 융합을 활발히 일으키는 시기와 거의 같았다. Myoglobin은 분화초기부터 서서히 합성축적되기 시작하여 배양 5일에서 최대치에 달하였다. Creatine phosphokinase는 배양 3일만에, 그리고 glyceraldehyde dehydrogenase는 6일만에 전기영동상에 검출되었다. Tropomyosin $\\alpha$와 $\\beta$, 그리고 troponin C는 분화초기부터 비교적 다량 합성되고 있었다. 젖산탈수소효소의 활성은 배양 2$\\sim$5일 사이에서 급격히 증가하고 이후 거의 변화가 없었다. 이 효소의 동위효소 조성은 초기 근원세포에서는 $H_4$와 $H_3M$형이 많으나 분화가 진행됨에 따라 $HM_3 와 M_4$형이 서서히 출현하였다. 그리고 배양 5일만에 5종의 동위효소가 모두 검출되었다.

• Journal of Life Science
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• v.19 no.2
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• pp.256-263
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• 2009

The lactate dehydrogenase (EC 1.1.1.27, LDH) $A_4$ isozyme in skeletal muscle of mandrin fish (Siniperca scherzeri) was successfully purified by affinity chromatography and ultrafiltration. The molecular weight of the purified LDH $A_4$ isozyme was 140.4 kDa and its isoelectric point (pI) was 7.0. Optimal pH for enzymatic reaction was 7.5. ${K_m}^{PYR}$ and $V_{max}$ value of the purified LDH $A_4$ isozyme were $4.86{\times}10^{-5}$ M and 13.31 mM/min using pyruvate as a substrate, respectively. These kinetic properties of the purified LDH $A_4$ isozyme supported the fact that the mandrin fish was a warm-adapted species. The antibody against the purified LDH $A_4$ isozyme may be used in the metabolic physiological studies of ectothermic vertebrates and in the diagnosis of several human diseases.

• Journal of Life Science
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• v.26 no.2
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• pp.155-163
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• 2016

Metabolism of lactate dehydrogenase (EC 1.1.1.27, LDH) was studied to identify the function of LDH-C. Tissues of LDH liver-specific Ldh-C expressed Carassius auratus and eye-specific Ldh-C expressed Lepomis macrochirus after starvation were studied. LDH activity in liver tissue from C. auratus was increased after starvation. And LDH specific activity (units/mg) and LDH/CS were increased in tissues. It means the anaerobic metabolism was taking place in C. auratus after starvation. LDH B₄ isozyme was decreased in skeletal muscle and increased in heart tissue. LDH C₄ isozymes those showed in eye and brain tissues were identified as liver-specific C₄ isozymes and disappeared after starvation. And C hybrid in eye, A₄ isozyme in brain, and both C hybrid and C₄ isozyme in liver tissue were increased, respectively. In L. macrochirus, the level of variation of LDH activities was low but greatly increased especially in eye tissue and LDH A₄ and AC hybrid were increased in brain tissue. The LDH activities in tissues from C. auratus and L. macrochirus remained 30.30-18.64% and 25-18.75%, respectively, as a result of the inhibition by 10 mM of pyruvate. The Km^PYR values of LDH in C. auratus were increased. As a result, LDH liver-specific C₄ isozyme was expressed in liver, brain and eye tissues during starvation. It seems metabolism of lactate was predominant in brain tissue. After starvation, the liver-specific LDH-C was affected more than eye-specific LDH-C.

• 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.

• 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.

• Korean Journal of Environmental Biology
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• v.17 no.3
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• pp.263-270
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• 1999

In order to identify the morphological changes of tissues in mouse after irradiation. We have observed the redistribution of LDH isozymes and the morphological changes of skeletal muscle, heart, kidney, liver and testis in mouse according to variation amount with the time after the 1 Gray and 3 Gray irradiation each. As a result of H-E (hematoxylin-eosin) stain, the apoptotic bodies were more easily observed in the liver than the other tissues and the quantity of the apoptotic bodies was proportionated to radiation amount. The number of apoptotic bodies was shown the highest at 1 day in most tissues and at 7 day in testis after irradiation. TUNEL (terminal deoxyribonucleodtidyl transferase-mediated dUTP-digoxigenin nick end labeling) staining was shown the same results as H-E staining. After the irradiation, the protein content was reduced in tissues except kidney. And protein content was reduced in all tissues at the initial period of 2 hours after 3 Gy irradiation. But it increased at 7 days after irradiation. LDH (EC 1.1.1.27, lactate dehydrogenase) activity was increased mostly in tissues at the early stage after 1 Gy irradiation. The maximum activity was detected earlier stage after 1 Gy irradiation than 3 Gy irradiation. The activity of LDH $A_4$ isozyme was decreased in the skeletal muscle, heart, kidney, and testis. The activity of $B_4$ and $A_2$$B_2$ sozyme was increased in the skeletal muscle and heart, and the activity of heterotetramer isozyme was increased in kidney The activity of $A_4$ isozyme in liver was detected high level and the activity of isozyme including subunit C elevated in testis. Therefore, LDH isozyme seems to play a role of lactate oxidase in most tissues except liver after irradiation. These data support that LDH isozyme is predomintly involved in the aerobic metabolism.

• Journal of Life Science
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• v.28 no.1
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• pp.1-8
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• 2018

To confirm the function of lactate dehydrogenase (LDH) (EC 1.1.1.27, LDH), its metabolism was studied by activity, kinetics, and isozyme analysis in tissues of Ldh testis-specific C expressing mice (Mus musculus) maintained in a state of starvation for 48 hr and 96 hr. In skeletal muscle, liver, and eye tissues, LDH and LDH $A_4$ activity increased and anaerobic metabolism predominated. While LDH activity in the heart and kidney tissues decreased, LDH $B_4$ activity increased and aerobic metabolism predominated, producing pyruvic acid. In the testis tissue, LDH $C_4$ activity decreased. In the brain tissue, LDH activity increased, but the isozyme change was small and the amount of pyruvic acid decreased. $K{_m}^{PYR}$ increased in tissues other than kidney tissue, and the affinity for pyruvic acid decreased. Consequently, in Ldh-A and B-expressing tissues, the activities of isozymes with higher concentrations increased. However, in Ldh-A, B, and C-expressing tissue, $C_4$ decreased and the function of the tissue also decreased. In particular, LDH in brain tissue played a role as a pyruvate reductase. Therefore, this process might be the mechanism for producing energy in the state of starvation.

• Journal of Life Science
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• v.31 no.12
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• pp.1066-1071
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• 2021

The aim of this study was to examine the metabolic adjustment of lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes to a change in dissolved oxygen (DO) in bluegill (Lepomis macrochirus). After bluegills were adapted to a constant environment in an aquarium, the DO was changed to investigate the activity of LDH isozyme and the relative ratio of subunits A, B, and C for each tissue. When the DO was decreased from 18 ppm to 6 ppm, LDH in skeletal muscle, heart, and brain tissues recovered to the level of control activity within 12, 12, and 6 hr, respectively. LDH activity changed in accordance with a change in DO. The compensation was performed rapidly and is thought to be an important function of LDH in enabling bluegills to adapt to their environment. In bluegill heart, eye, and brain tissues, the relative ratio of subunit A increased and showed a tendency to recover similarly to the subunit ratio of control groups up to 12 hr. It is thought that the anaerobic metabolism using subunit A was increased in the initial stage when DO was changed. In addition, the results revealed that subunit C was more similar to subunit A than subunit B. In bluegills, subunits A and C of LDH seem to be evolutionarily similar. LDH isozymes, mainly containing subunits A and C, are likely responsible for the function of pyruvate reductase, which plays a role in making the bluegill adapt to a hypoxic environment through anaerobic metabolism.

• Journal of Life Science
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• v.22 no.2
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• pp.209-219
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• 2012

The properties of lactate dehydrogenase (LDH, EC 1.1.1.27) eye-specific $C_4$ isozyme were studied by polyacrylamide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme kinetics. Furthermore, we proposed the optimal conditions for measuring the activity of LDH eye-specific $C_4$ isozyme. The isozymes were detected in the cytosol of eye tissues from Lepomis macrochirus and Micropterus salmoides and were more similar to the $A_4$ than the $B_4$ isozyme. LDH/CS in the eye tissue of L. macrochirus was increased in September, so the ratio of anaerobic metabolism was high. The electrophoretic patterns of mitochondrial LDH were similar to those of cytosolic LDH in the eye tissues of L. macrochirus and Micropterus salmoides. LDH eye-specific $C_4$ isozyme from eye tissue was purified by preparative native-PAGE. The activities of LDH eye-specific $C_4$ isozymes in L. macrochirus and M. salmoides were reduced at concentrations greater than 0.2 mM and 0.1 mM of pyruvate, respectively. These concentrations remained at 5.2% and 15.8% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The LDH activities of eye tissues were reduced at concentrations greater than 22 mM and 24 mM of lactate, respectively, in L. macrochirus and M. salmoides. The ${K_m}^{PYR}$ of eye-specific $C_4$ was 0.088 mM in L. macrochirus and it was 0.033 mM in M. salmoides. The activities of cytosolic and mitochondrial eye-specific $C_4$ isozymes were high in ${\alpha}$-ketobutyric acid. Furthermore, the activities of eye tissue and eye-specific $C_4$ isozyme had to be measured with 0.5 mM of pyruvate and a buffer solution of pH 7.5. As a conclusion, the eye-specific $C_4$ isozyme in M. salmoides has a high affinity for pyruvate and exhibits maximum activity at a lower concentration of pyruvate and at higher concentration of lactate than that in L. macrochirus. Therefore, it seems that the energy produced by the LDH eye-specific $C_4$ isozyme in M. salmoides was used at the first stage of predatory behavior.

• 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.