Analysis of glutathione S-transferase-catalyzed S-alkylglutathione formation by high-performance liquid chromatography.
Analysis of glutathione S-transferase-catalyzed S-alkylglutathione formation by high-performance liquid chromatography.
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通过高效液相色谱分析谷胱甘肽 S-转移酶催化的 S-烷基谷胱甘肽形成。
DOI：
10.1016/0003-2697(91)90034-q
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发表时间：
1991
影响因子：
2.9
通讯作者：
O'Leary,KA
中科院分区：
文献类型：
--
作者：
Tracy,JW;O'Leary,KA
通过谷胱甘肽S-转移酶（EC 2.5.1.18）代谢烷基卤化物和某些有机磷化合物，导致形成常见产物S-烷基谷胱甘肽。我们已经开发了一种高效液相色谱法测定S-甲基谷胱甘肽和S-乙基谷胱甘肽的形成，适用于测量对各种异生质底物的酶活性。将缀合物用1-氟-2，4-二硝基苯衍生化以形成相应的N-2，4-二硝基苯衍生物，然后通过具有梯度洗脱的反相HPLC分离。该方法的实用性是通过使用部分纯化的制备大鼠肝脏谷胱甘肽S-转移酶和几个原型基板，包括碘甲烷，碘乙烷，敌敌畏，和甲基敌敌畏。N-（2，4-二硝基苯基）-S-烷基谷胱甘肽的检测限约为50 pmol。该方法的优点比其他测定的S-烷基转移酶活性进行了讨论。
Metabolism of alkyl halides and some organophosphorous compounds by glutathione S-transferases (EC 2.5.1.18) leads to formation of an S-alkylglutathione as a common product. We have developed an HPLC assay for formation of S-methylglutathione and S-ethylglutathione that is applicable to measuring enzyme activity toward a variety of xenobiotic substrates. The conjugates are derivatized with 1-fluoro-2,4-dinitrobenzene to form the corresponding N-2,4-dinitrophenyl derivatives, which are then separated by reverse-phase HPLC with gradient elution. The utility of the method is illustrated by the use of partially purified preparations of rat liver glutathione S-transferases and several prototypic substrates including iodomethane, iodoethane, dichlorvos, and methyl parathion. The limit of detection is about 50 pmol of N-(2,4-dinitrophenyl)-S-alkylglutathione. Advantages of the method over other assays of S-alkyl transferase activity are discussed.