Stabilization of purified human 5-lipoxygenase with glutathione peroxidase and superoxide dismutase.
Stabilization of purified human 5-lipoxygenase with glutathione peroxidase and superoxide dismutase.
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使用谷胱甘肽过氧化物酶和超氧化物歧化酶稳定纯化的人 5-脂氧合酶。
DOI：
10.1006/abio.1994.1294
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发表时间：
1994
影响因子：
2.9
通讯作者：
B. Samuelsson
中科院分区：
文献类型：
--
作者：
Y. Y. Zhang;M. Hamberg;O. Rådmark;B. Samuelsson
人5-脂氧合酶（5LO）在纯化后变得非常不稳定。常用的蛋白质稳定化方法不能防止这种失活。然而，向纯化的5LO溶液（300-500微克/毫升）中加入少量谷胱甘肽过氧化物酶（0.15微克/毫升）和超氧化物歧化酶（1微克/毫升）可使酶在储存期间稳定。受保护的5LO在25 ℃下保持完全活性至少12天，而在没有保护的情况下，50%的活性在10小时内丧失。单独的谷胱甘肽过氧化物酶也保留了5-脂氧合酶的活性，然而，在没有超氧化物歧化酶的情况下，效果迅速下降。2-巯基乙醇是谷胱甘肽过氧化物酶保护5LO的最有效的供氢底物。过氧化氢酶作为稳定剂的有效性较低，而合成的谷胱甘肽过氧化物酶模拟化合物依布硒啉（ebselen）并不能保护5LO。由于许多金属离子结合蛋白对H2 O2失活敏感，因此该方法也可用于其他蛋白的稳定。
Human 5-lipoxygenase (5LO) becomes very unstable after purification. Commonly used methods for protein stabilization could not prevent this inactivation. However, addition of small amounts of glutathione peroxidase (0.15 micrograms/ml) and superoxide dismutase (1 microgram/ml) to the solution of purified 5LO (300-500 micrograms/ml) stabilized the enzyme during storage. The protected 5LO maintained full activity for at least 12 days at 25 degrees C, while 50% of the activity was lost within 10 h without protection. Glutathione peroxidase alone also preserved the activity of 5-lipoxygenase; however, the effect declined rapidly in the absence of superoxide dismutase. 2-Mercaptoethanol was the most efficient hydrogen donor substrate for glutathione peroxidase in the protection of 5LO. Catalase was less effective as a stabilizing agent, and ebselen, a synthetic glutathione peroxidase-mimicking compound, did not protect 5LO. Since many metal ion binding proteins are susceptible to H2O2 inactivation, this method could be useful also for the stabilization of other proteins.