Determination of di–/polyamine oxidase activity in plants by an in-gel spermidine oxidation assay

Abstract

Diamine and polyamine catabolism controls plant development, resistance to pathogens and stress responses. Diamine and polyamine oxidases control the catabolism of diamines and polyamines, respectively. Two major routes of di–/polyamine catabolism exist: the terminal and the interconverting. The in vitro activity of each route is assayed by the colorimetric or chemiluminescent determination of hydrogen peroxide produced by oxidation of di–/polyamine substrates. However, these assays fail to estimate activity of individual di–/polyamine oxidase isoenzymes. Herein, I describe an assay for the simultaneous in-gel determination of terminal and interconverting di–/polyamine oxidase isoenzyme activities.