Smith I K, Vierheller T L, Thorne C A
Department of Botany, Ohio University, Athens 45701-2979.
Anal Biochem. 1988 Dec;175(2):408-13. doi: 10.1016/0003-2697(88)90564-7.
A method for assaying glutathione reductase (GSH; EC 1.6.4.2) in crude plant extracts is described. The method is based on the increase in absorbance at 412 nm when 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) is reduced by GSH. The effects of the following parameters on the assay were tested: various buffers, pH, buffer concentration, compounds commonly present in enzyme preparations, thiols, and the presence of another NADPH-dependent enzyme. The assay is more sensitive and less subject to interference than the widely used assay where NADPH oxidation is monitored. In particular, the specificity of DTNB allows assay of glutathione reductase in the presence of other NADPH-dependent enzymes and common protein extract contaminants.
本文描述了一种用于测定植物粗提物中谷胱甘肽还原酶（GSH；EC 1.6.4.2）的方法。该方法基于谷胱甘肽还原5,5'-二硫代双（2-硝基苯甲酸）（DTNB）时在412nm处吸光度的增加。测试了以下参数对该测定的影响：各种缓冲液、pH值、缓冲液浓度、酶制剂中常见的化合物、硫醇以及另一种依赖NADPH的酶的存在。与广泛使用的监测NADPH氧化的测定方法相比，该测定方法更灵敏且受干扰更少。特别是，DTNB的特异性使得在存在其他依赖NADPH的酶和常见蛋白质提取物污染物的情况下能够测定谷胱甘肽还原酶。
