Identification of novel PAMP-triggered phosphorylation and dephosphorylation events in arabidopsis thaliana by quantitative phosphoproteomic analysis
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Plant Science Program
Desert Agriculture Initiative
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AbstractSignaling cascades rely strongly on protein kinase-mediated substrate phosphorylation. Currently a major challenge in signal transduction research is to obtain high confidence substrate phosphorylation sites and assign them to specific kinases. In response to bacterial flagellin, a pathogen-associated molecular pattern (PAMP), we searched for rapidly phosphorylated proteins in Arabidopsis thaliana by combining multistage activation (MSA) and electron transfer dissociation (ETD) fragmentation modes, which generate complementary spectra and identify phosphopeptide sites with increased reliability. Of a total of 825 phosphopeptides, we identified 58 to be differentially phosphorylated. These peptides harbor kinase motifs of mitogen-activated protein kinases (MAPKs) and calcium-dependent protein kinases (CDPKs), as well as yet unknown protein kinases. Importantly, 12 of the phosphopeptides show reduced phosphorylation upon flagellin treatment. Since protein abundance levels did not change, these results indicate that flagellin induces not only various protein kinases but also protein phosphatases, even though a scenario of inhibited kinase activity may also be possible. © 2014 American Chemical Society.
SponsorsD.P. thanks the Agence Nationale pour la Recherche (ANR) for the funding ANR-2010-JCJC-1608. This work was supported by the CNRS, Genopole-France, Institut National de la Recherche Agronomique, Universite d'Evry Val d'Essonne and Region Ile-de-France. We are also indebted to Benoit Valot, Edlira Nano, Olivier Langella, and Michel Zivy for help with using MassChroQ,
PublisherAmerican Chemical Society (ACS)
JournalJournal of Proteome Research
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