14-3-3 proteins and a 13-lipoxygenase form associations in a phosphorylation-dependent manner.
Recently, we have demonstrated by two different methods that lipoxgenases (LOXs) and 14-3-3 proteins form interactions in barley embryos [Holtman, Roberts, Oppedijk, Testerink, van Zeijl and Wang (2000) FEBS Lett. 474, 48-52]. It was shown by both co-immunoprecipitations and surface-plasmon resonance experiments that 13-LOX, but not 9-LOX, forms interactions with 14-3-3 proteins. In the present report we show that the presence of 13-LOX and 14-3-3 proteins was established in high-molecular-mass complexes. Amounts of 13-LOX and 14-3-3 proteins in high-molecular-mass fractions increased during germination, but were reduced after dephosphorylation of protein extracts or competition with the 14-3-3-binding peptide P-Raf-259, indicating that 13-LOX and 14-3-3 proteins interact in a phosphorylation-dependent manner.
Holtman
W L
WL
Center for Phytotechnology UL-TNO, Department of Applied Plant Sciences, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands. holtman@voeding.tno.nl
Roberts
M R
MR
Wang
M
M
D048948:14-3-3 Proteins; D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D001467:Hordeum / Q000201:enzymology*; D008084:Lipoxygenase / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D012639:Seeds / Q000201:enzymology; D014446:Tyrosine 3-Monooxygenase / Q000302:isolation & purification / Q000378:metabolism*
D048948:14-3-3 Proteins; C117433:13-lipoxygenase; D008084:Lipoxygenase; D014446:Tyrosine 3-Monooxygenase