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首页> 外文期刊>Biochemistry >ROLE OF LIPID HYDROPEROXIDES IN THE ACTIVATION OF 15-LIPOXYGENASE
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ROLE OF LIPID HYDROPEROXIDES IN THE ACTIVATION OF 15-LIPOXYGENASE

机译：脂质过氧化物在15-脂氧合酶激活中的作用

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摘要

We have used stopped-flow rapid reaction methods, employing both fluorescence and absorbance monitoring, together with HPLC analysis of the products to study the activation of soybean 15-lipoxygenase by 13(S)-hydroperoxy-9,11(E,Z)-octadecadienoic acid (13-HPOD), When lipoxygenase is mixed with an equimolar concentration of 13-HPOD, the enzyme undergoes a rapid change in fluorescence. The rate of the change of fluorescence is dependent on the concentration of the 13-HPOD (k = 6.7 x 10(6) M(-1) s(-1)) and is accompanied by activation of the enzyme. The fluorescence change is not accompanied by any change in the UV absorbance of the 13-HPOD, suggesting no loss of the conjugated diene during enzyme activation, and HPLC analysis of the products of the reaction confirms that the 13-HPOD can be recovered unchanged following this reaction. In the presence of an inhibitor (BWA4C, a hydroxamate inhibitor) that reduces the active-site iron, the 13-HPOD and the inhibitor are destroyed in a peroxidase-like reaction. On the basis of these observations we propose that 13-HPOD binds to the enzyme and facilitates activation of the enzyme, possibly through the formation of a protein radical, and that the 13-HPOD is not changed chemically in this process.
机译：我们已经使用了停流快速反应方法，同时进行了荧光和吸光度监测，以及产品的HPLC分析，以研究13（S）-hydroperoxy-9,11（E，Z）-对大豆15-脂加氧酶的活化作用。十八碳二烯酸（13-HPOD），当脂氧合酶与等摩尔浓度的13-HPOD混合时，该酶的荧光快速变化。荧光的变化速率取决于13-HPOD的浓度（k = 6.7 x 10（6）M（-1）s（-1）），并伴有酶的激活。荧光变化不伴随13-HPOD的紫外线吸收变化，表明酶活化过程中共轭二烯没有损失，反应产物的HPLC分析证实13-HPOD可以在回收后保持不变这个反应。在还原活性位点铁的抑制剂（BWA4C，异羟肟酸酯抑制剂）的存在下，过氧化物酶样反应会破坏13-HPOD和抑制剂。基于这些观察结果，我们提出13-HPOD可能会通过形成蛋白质自由基而与酶结合并促进酶的活化，并且在此过程中13-HPOD不会发生化学变化。

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