Sensitivity of plant mitochondrial terminal oxidases to the lipid peroxidation product 4-hydroxy-2-nonenal (HNE).
We have investigated the effect of the lipid peroxidation product, HNE (4-hydroxy-2-nonenal), on plant mitochondrial electron transport. In mitochondria isolated from Arabidopsis thaliana cell cultures, HNE inhibited succinate-dependent oxygen consumption via the Aox (alternative oxidase), but had minimal effect on respiration via Cox (cytochrome c oxidase). Maximal Cox activity, measured with reduced cytochrome c as substrate, was only slightly inhibited by high concentrations of HNE, at which Aox was completely inhibited. Incubation with HNE prevented dimerization of the Aox protein, suggesting that one site of modification was the conserved cysteine residue involved in dimerization and activation of this enzyme (Cys(I)). However, a naturally occurring isoform of Aox lacking Cys(I) and unable to be dimerized, LeAox1b from tomato (Lycopersicon esculentum), was equally sensitive to HNE inhibition, showing that other amino acid residues in Aox also interact with HNE. The presence of HNE in vivo in Arabidopsis cell cultures was also investigated. Induction of oxidative stress in the cell cultures by the addition of hydrogen peroxide, antimycin A or menadione, caused a significant increase in hydroxyalkenals (of which HNE is the most prominent). Western blotting of mitochondrial proteins with antibodies against HNE adducts, demonstrated significant modification of proteins during these treatments. The implications of these results for the response of plants to reactive oxygen species are discussed.
Winger
Alison M
AM
Biochemistry and Molecular Biology, School of Biomedical and Chemical Sciences, The University of Western Australia, Crawley 6009, WA, Australia.
Millar
A Harvey
AH
Day
David A
DA
D000447:Aldehydes / Q000378:metabolism*; D017360:Arabidopsis; D029681:Arabidopsis Proteins / Q000378:metabolism; D002478:Cells, Cultured; D015227:Lipid Peroxidation; D008928:Mitochondria / Q000201:enzymology*; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000378:metabolism*; D010101:Oxygen Consumption; D020033:Protein Isoforms
D000447:Aldehydes; D029681:Arabidopsis Proteins; D020033:Protein Isoforms; C462359:tert-4-hydroxy-2-nonenal; D010088:Oxidoreductases