Uses
Glutathione S-Transferase from human placenta has been used in antigen microarray and informatics analysis.
General Description
Glutathione S-transferase (GST) is a major detoxification enzyme, and exists as multiple cytoplasmic and membrane-bound isozymes. These isozymes differ in their catalytic activity, as well as in their non-catalytic binding properties. Cytoplasmic isoforms of GST are encoded by five genes, namely α, θ, μ, σ and π. α, μ and π are the most abundant forms in mammals. Membrane bound GST forms are encoded by a single gene.
Biochem/physiol Actions
Glutathione S-transferase (GST) catalyzes conjugation reactions during phase II metabolism. This enzyme also confers protection against catabolic products of peroxidized lipid and oxidized DNA, and ROS (reactive oxygen species) or electrophiles.
Purification Methods
It is purified by affinity chromatography using a column prepared by coupling glutathione to epoxy-saturated Sepharose. After washing contaminating proteins, the pure transferase is eluted with buffer containing reduced glutathione. The solution is then concentrated by ultrafiltration, dialysed against phosphate buffer at pH ~7 and stored in the presence of dithiothreitol (2mM) in aliquots at <-20o. [Simons & Vander Jag Anal Biochem 52 334 1977.]
