5 alpha,6 alpha-Epoxy-cholestan-3 beta-ol (cholesterol alpha-oxide): A specific substrate for rat liver glutathione transferase B.

A semi-micro assay was developed for the conjugation of 5 alpha,6 alpha-epoxy-cholestan-3 beta-ol (cholesterol alpha-oxide) with glutathione. The soluble supernatant of rat liver homogenate catalysed the reaction at a rate of 0.2-0.5 pmol . min-1 . mg protein-1 with 4 microM cholesterol alpha-oxide, while the reaction in the presence of GSH alone was barely detectable. Enzymic activity in the soluble supernatant was due equally to the two forms of glutathione transferase B (approximately 100 pmol . min-1 . mg protein-1), glutathione transferases AA, A, C and E being unreactive. The activity of purified glutathione transferase B was about 5-times that expected from the activity of the soluble supernatant. Complex enzyme kinetics were obtained suggestive of substrate inhibition.