Determination of di-/Polyamine Oxidase Activity in Plants by an In-Gel Spermidine Oxidation Assay

Methods Mol Biol. 2018;1694:141-147. doi: 10.1007/978-1-4939-7398-9_14.

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.

Keywords: Diamine oxidases; Flavin adenine dinucleotide; Hydrogen peroxide; Oxidation; Peroxisomes; Polyamine back-conversion; Polyamine interconversion; Polyamine oxidases; Spermidine.

MeSH terms

  • Enzyme Assays*
  • Hydrogen Peroxide / chemistry
  • Hydrogen Peroxide / metabolism
  • In Vitro Techniques
  • Oxidation-Reduction*
  • Oxidoreductases Acting on CH-NH Group Donors / metabolism*
  • Plant Extracts / chemistry
  • Plants / enzymology*
  • Spermidine / chemistry*

Substances

  • Plant Extracts
  • Hydrogen Peroxide
  • Oxidoreductases Acting on CH-NH Group Donors
  • polyamine oxidase
  • Spermidine