Novel control of lactate dehydrogenase from the freeze tolerant wood frog: role of posttranslational modifications.
Novel control of lactate dehydrogenase from the freeze tolerant wood frog: role of posttranslational modifications.
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作者：
Abboud J;Storey KB
乳酸脱氢酶（LDH）是无氧糖酵解的末端酶，在氧限制条件下维持糖酵解ATP的产生和在应激条件缓解时促进积累的乳酸的催化中起着至关重要的作用。在这项研究中，在体内冷冻和脱水应力（这两个施加缺氧/缺氧应力的组织）对LDH的影响进行了研究，在骨骼肌的抗冻性林蛙，林蛙。从对照、冷冻和脱水的林蛙肌肉中纯化LDH，以两步方法纯化至均一。对纯化后的LDH进行动力学性质和稳定性分析，发现冷冻前后的Vmax、Km和I_（50）值无显著性差异。然而，控制和脱水LDH显着不同的Km值丙酮酸，乳酸，和NAD，I50尿素，并在温度，葡萄糖和尿素对这些参数的影响。LDH的翻译后修饰的可能性是负责控制和脱水状态之间的酶行为的稳定差异进行了评估，使用ProQ钻石染色检测磷酸化和免疫印迹检测乙酰化，甲基化，泛素化，SUMO化和亚硝基化的酶。LDH从肌肉脱水林蛙表现出显着较低的乙酰化水平，提供了一个潜在的作用，蛋白质乙酰化的应激反应控制的代谢酶的第一个演示。
Lactate dehydrogenase (LDH), the terminal enzyme of anaerobic glycolysis, plays a crucial role both in sustaining glycolytic ATP production under oxygen-limiting conditions and in facilitating the catabolism of accumulated lactate when stress conditions are relieved. In this study, the effects on LDH of in vivo freezing and dehydration stresses (both of which impose hypoxia/anoxia stress on tissues) were examined in skeletal muscle of the freeze-tolerant wood frog, Rana sylvatica. LDH from muscle of control, frozen and dehydrated wood frogs was purified to homogeneity in a two-step process. The kinetic properties and stability of purified LDH were analyzed, revealing no significant differences in Vmax, Km and I50 values between control and frozen LDH. However, control and dehydrated LDH differed significantly in Km values for pyruvate, lactate, and NAD, I50 urea, and in temperature, glucose, and urea effects on these parameters. The possibility that posttranslational modification of LDH was responsible for the stable differences in enzyme behavior between control and dehydrated states was assessed using ProQ diamond staining to detect phosphorylation and immunoblotting to detect acetylation, methylation, ubiquitination, SUMOylation and nitrosylation of the enzyme. LDH from muscle of dehydrated wood frogs showed significantly lower levels of acetylation, providing one of the first demonstrations of a potential role for protein acetylation in the stress-responsive control of a metabolic enzyme.
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影响因子：
--
作者：
CHURCHILL, TA;STOREY, KB
通讯作者：
STOREY, KB
DOI：
10.1007/s003600050078
发表时间：
1997-05-01
期刊：
JOURNAL OF COMPARATIVE PHYSIOLOGY B-BIOCHEMICAL SYSTEMIC AND ENVIRONMENTAL PHYSIOLOGY
影响因子：
--
作者：
Holden, CP;Storey, KB
通讯作者：
Storey, KB
DOI：
10.1201/9780203647073.ch7
发表时间：
2004-01-01
期刊：
LIFE IN THE FROZEN STATE
影响因子：
--
作者：
Storey, KB;Storey, JM
通讯作者：
Storey, JM
影响因子：
21.3
作者：
Cohen, P
通讯作者：
Cohen, P
影响因子：
2.7
作者：
Biggar, Kyle K.;Dawson, Neal J.;Storey, Kenneth B.
通讯作者：
Storey, Kenneth B.