DETERMINATION OF GLUTATHIONE AND GLUTATHIONE DISULFIDE USING GLUTATHIONE-REDUCTASE AND 2-VINYLPYRIDINE
DETERMINATION OF GLUTATHIONE AND GLUTATHIONE DISULFIDE USING GLUTATHIONE-REDUCTASE AND 2-VINYLPYRIDINE
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DOI：
10.1016/0003-2697(80)90139-6
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发表时间：
1980-01-01
影响因子：
2.9
通讯作者：
GRIFFITH, OW
中科院分区：
文献类型：
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作者：
GRIFFITH, OW
生物样品的总谷胱甘肽含量是方便地确定与酶循环测定的基础上谷胱甘肽还原酶。在该方法的原始和后续几个描述中，通过测定样品（其中谷胱甘肽通过N-乙基马来酰亚胺预处理掩蔽）选择性测定谷胱甘肽二硫化物。由于N-乙基马来酰亚胺是谷胱甘肽还原酶的有效抑制剂，因此有必要去除过量的试剂;所使用的程序是费力的，并且显著地导致实验误差。2-乙烯基吡啶是一种较好的谷胱甘肽衍生化试剂。与N-乙基马来酰亚胺相反，2-乙烯基吡啶不会显著抑制谷胱甘肽还原酶，因此不需要从样品溶液中除去。2-乙烯基吡啶在微酸性pH值下与谷胱甘肽反应，此时谷胱甘肽二硫化物的自发形成最小。小鼠血浆中的总谷胱甘肽浓度显著高于通常报告的浓度，谷胱甘肽二硫化物占总谷胱甘肽的比例低于30%。
The total glutathione content of biological samples is conveniently determined with an enzymatic recycling assay based on glutathione reductase. In the original and several subsequent descriptions of this procedure, glutathione disulfide is selectively determined by assaying samples in which glutathione is masked by pretreatment with N-ethylmaleimide. Since N-ethylmaleimide is a potent inhibitor of glutathione reductase, it is necessary to remove excess reagent; the procedures used are laborious and contribute significantly to experimental error. 2-Vinyl-pyridine is a much better reagent for the derivitization of glutathione. In contrast to N-ethylmaleimide, 2-vinylpyridine does not inhibit glutathione reductase significantly and therefore need not be removed from the sample solutions. 2-Vinylpyridine reacts with glutathione at slightly acidic pH values where spontaneous formation of glutathione disulfide is minimal. The total glutathione concentration in mouse plasma is substantially higher than generally reported and glutathione disulfide constitutes less than 30% of the total glutathione present.