Spectrophotometric assay for serum glutathione transferase: A re-examination

详细信息	查看全文 | 推荐本文 |

• 作者：Raffaele Fabrini^a ; ¹ ; Alessio Bocedi^a ; ¹ ; Renato Massoud^b ; Giorgio Federici^b ; Giorgio Ricci^a ; ^{riccig@uniroma2.it}
• 关键词：Glutathione transferase assay ; Serum glutathione transferase ; Artifact in GST activity ; Blood GST
• 出版年：2012
• 期刊代码：55_00099120
• 类别：med
• 出版时间：June, 2012
• 卷：45
• 期：9
• 页码：668-671
• 文件大小：266 K

摘要

Objectives

The aim of the present paper is a careful re-examination of an automated spectrophotometric procedure for glutathione transferase (GST) activity in human serum described previously and used in many laboratories.

Design and methods

GST activity in human serum has been assayed spectrophotometrically under various experimental conditions. Recombinant human GSTs and specific inhibitors were also used to check the possible occurrence of artifacts.

Results

Basal level of the enzyme calculated using this method turns out to be much higher than that found using RIA and ELISA procedures. Relevant pH-dependent artifacts deeply affect this spectrophotometric assay. Notably, spectral changes previously interpreted as a measure of basal activity, are mainly due to an increase of the spontaneous reaction between the two substrates.

Conclusion

GST activity in normal serum cannot be correctly determined with the spectrophotometric assay described previously because of the very low enzyme concentration and the pH-dependent artifacts.