Methods for purification of glutathione peroxidase and related enzymes.

ABSTRACT

The different preparative techniques and related analytical methods used for purification of glutathione peroxidase, glutathione transferase and glutathione reductase, described in papers published in the last ten years, have been reviewed in this article. Among the different purification techniques, chromatography has played a relevant role, being reported in all the papers reviewed, whereas other preparative techniques such as electrophoresis and isoelectric focusing were less employed and have been reported in only ca. 3% of cases. Frequently, several different chromatographic modes and several rechromatography steps have been employed. The use of at least three different chromatographic modes has been reported in 53% of total reviewed papers, whereas 41% of them employed two differents modes and in only 6% a single preparative chromatographic step was used. To evaluate losses and improve recovery, analytical methods for quantitation of protein and assay of enzymatic activity must be used in each purification step. Among these analytical techniques, gel electrophoresis, under denaturing conditions, has been widely used to assess purity of enzyme preparation. A discussion of the different activity assay methods used for these three enzymes is also presented in this article.