Publication Information
Vu et al., 2016
Abstract
J Proteome Res. 2016 Dec 2;15(12):4304-4317. doi: 10.1021/acs.jproteome.6b00348.
Epub 2016 Oct 4.
Up-to-Date Workflow for Plant (Phospho)proteomics Identifies Differential
Drought-Responsive Phosphorylation Events in Maize Leaves.
Vu LD(1)(2)(3)(4), Stes E(1)(2)(3)(4), Van Bel M(1)(2), Nelissen H(1)(2),
Maddelein D(3)(4), Inzé D(1)(2), Coppens F(1)(2), Martens L(3)(4), Gevaert
K(3)(4), De Smet I(1)(2).
Author information:
(1)Department of Plant Systems Biology, VIB , 9052 Ghent, Belgium.
(2)Department of Plant Biotechnology and Bioinformatics, Ghent University , 9052
Ghent, Belgium.
(3)Medical Biotechnology Center, VIB , 9000 Ghent, Belgium.
(4)Department of Biochemistry, Ghent University , 9000 Ghent, Belgium.
Protein phosphorylation is one of the most common post-translational
modifications (PTMs), which can regulate protein activity and localization as
well as protein-protein interactions in numerous cellular processes.
Phosphopeptide enrichment techniques enable plant researchers to acquire insight
into phosphorylation-controlled signaling networks in various plant species.
Most phosphoproteome analyses of plant samples still involve stable isotope
labeling, peptide fractionation, and demand a lot of mass spectrometry (MS)
time. Here, we present a simple workflow to probe, map, and catalogue plant
phosphoproteomes, requiring relatively low amounts of starting material, no
labeling, no fractionation, and no excessive analysis time. Following
optimization of the different experimental steps on Arabidopsis thaliana
samples, we transferred our workflow to maize, a major monocot crop, to study
signaling upon drought stress. In addition, we included normalization to protein
abundance to identify true phosphorylation changes. Overall, we identified a set
of new phosphosites in both Arabidopsis thaliana and maize, some of which are
differentially phosphorylated upon drought. All data are available via
ProteomeXchange with identifier PXD003634, but to provide easy access to our
model plant and crop data sets, we created an online database, Plant PTM Viewer
( bioinformatics.psb.ugent.be/webtools/ptm_viewer/ ), where all phosphosites
identified in our study can be consulted.
DOI: 10.1021/acs.jproteome.6b00348
PMID: 27643528 [Indexed for MEDLINE]
