Journal Article

Bridging the Gap between Plant and Mammalian Polyamine Catabolism: A Novel Peroxisomal Polyamine Oxidase Responsible for a Full Back-Conversion Pathway in Arabidopsis

Roubelakis-Angelakis, Kalliopi A, Sanmartin, Maite, Moschou, Panagiotis N, Andriopoulou, Athina H, Sanchez-Serrano, Jose J, Rojo, Enrique

2008

Overview

In contrast to animals, where polyamine (PA) catabolism efficiently converts spermine (Spm) to putrescine (Put), plants have been considered to possess a PA catabolic pathway producing 1,3-diaminopropane, Δ¹-pyrroline, the corresponding aldehyde, and hydrogen peroxide but unable to back-convert Spm to Put. Arabidopsis (Arabidopsis thaliana) genome contains at least five putative PA oxidase (PAO) members with yet-unknown localization and physiological role(s). AtPAO1 was recently identified as an enzyme similar to the mammalian Spm oxidase, which converts Spm to spermidine (Spd). In this work, we have performed in silico analysis of the five Arabidopsis genes and have identified PAO3 (AtPAO3) as a nontypical PAO, in terms of homology, compared to other known PAOs. We have expressed the gene AtPAO3 and have purified a protein corresponding to it using the inducible heterologous expression system of Escherichia coli. AtPAO3 catalyzed the sequential conversion/oxidation of Spm to Spd, and of Spd to Put, thus exhibiting functional homology to the mammalian PAOs. The best substrate for this pathway was Spd, whereas the N¹-acetyl-derivatives of Spm and Spd were oxidized less efficiently. On the other hand, no activity was detected when diamines (agmatine, cadaverine, and Put) were used as substrates. Moreover, although AtPAO3 does not exhibit significant similarity to the other known PAOs, it is efficiently inhibited by guazatine, a potent PAO inhibitor. AtPAO3 contains a peroxisomal targeting motif at the C terminus, and it targets green fluorescence protein to peroxisomes when fused at the N terminus but not at the C terminus. These results reveal that AtPAO3 is a peroxisomal protein and that the C terminus of the protein contains the sorting information. The overall data reinforce the view that plants and mammals possess a similar PA oxidation system, concerning both the subcellular localization and the mode of its action.

• Publisher: American Society of Plant Biologists,American Society of Plant Physiologists
• Subject: Abscisic Acid / Abscisic Acid - pharmacology / agmatine / aldehydes / Amino Acid Sequence / analysis / Animals / Arabidopsis / Arabidopsis - enzymology / Arabidopsis - genetics / Arabidopsis - ultrastructure / Arabidopsis Proteins / Arabidopsis Proteins - chemistry / Arabidopsis Proteins - isolation & purification / Arabidopsis Proteins - physiology / Arabidopsis thaliana / Biochemical Processes and Macromolecular Structures / Biological and medical sciences / cadaverine / Catabolism / chemistry / Cloning, Molecular / Cyclopentanes / Cyclopentanes - pharmacology / drug effects / Enzymes / enzymology / Escherichia coli / Flavin-Adenine Dinucleotide / Flavin-Adenine Dinucleotide - metabolism / Fluorescence / Fundamental and applied biological sciences. Psychology / genes / Genes. Genome / genetics / green fluorescent protein / Green Fluorescent Proteins / Green Fluorescent Proteins - analysis / hydrogen peroxide / isolation & purification / Kinetics / mammals / Mammals - metabolism / mechanism of action / Messenger RNA / metabolism / Molecular and cellular biology / Molecular genetics / Molecular Sequence Data / Oxidases / Oxidation / Oxidoreductases Acting on CH-NH Group Donors / Oxidoreductases Acting on CH-NH Group Donors - chemistry / Oxidoreductases Acting on CH-NH Group Donors - isolation & purification / Oxidoreductases Acting on CH-NH Group Donors - physiology / Oxylipins / Oxylipins - pharmacology / Peroxisomes / Peroxisomes - drug effects / Peroxisomes - enzymology / pharmacology / Phylogeny / physiology / Plant Growth Regulators / Plant Growth Regulators - pharmacology / Plants / Polyamine Oxidase / Polyamines / Polyamines - metabolism / Proteins / putrescine / Recombinant Fusion Proteins / Recombinant Fusion Proteins - analysis / Sequence Alignment / spermine / ultrastructure