북방산개구리(Rana dybowskii) 조직 젖산탈수소효소 동위효소의 대사 조정 = Metabolic Adjustment of Lactate Dehydrogenase Isozymes in Tissues from Rana dybowskii

다국어 초록 (Multilingual Abstract)

The metabolic adjustment of LDH was investigated by quantifying lactate dehydrogenase (EC.1.1.1.27, LDH) activity and protein and characterizing purified LDH A4 isozyme in Rana sybowskii tissues collected in August. The LDH activity in skeletal muscle, heart, liver, eye and brain tissue was found to be particularly high at 1196.10, 231.50, 119.61, 50.48 and 76.13 units and the amount of protein was low at 15.21, 10.60, 24.57, 22.40 and 4.32 mg. Therfore the specific activity of LDH was high at 21.79, 4.87, 2.25, and 17.62 units/mg, respectively. Native- polyacrylamide electrophoresis showed stronger identification of LDH A4 isozyme in skeletal muscle and heart tissue. And A4 isozyme was slightly stronger in brain tissue, B4 isozyme weakened and eye-C hybrid appeared on the anode side. In the skeletal muscle, liver, and eye tissues, several bands appeared on the most negative side and are thought to be liver-specific C. The LDH A4 isozyme was purified by affinity chromatography at 65.6-72 ml after NAD+ entry, and all LDH isozymes were purified 6158.11 units with a yield of 73.54%. The LDH A4 was confirmed to be purified by SDS-PAGE, and the molecular weight of the subunit A was 29.9 kDa. The purified LDH A4 isozyme showed maximal activity in 0.5 mM pyruvic acid and then decreased in activity to 13.92% in 10 mM pyruvic acid. Therefore, KmPYR and VmaxPYR were 0.11 mM and 97.76 units, respectively. Therefore, although the affinity of LDH A4 to pyruvic acid was lowered slightly due to the increase in the temperature of the environment, it was considered that the LDH activity was greatly increased, and the anaerobic metabolism using pyruvic acid for energy generation was greatly increased.