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Analysis of the defence phosphoproteome of Arabidopsis thaliana using differential mass tagging
Author(s) -
Jones Alexandra M. E.,
Bennett Mark H.,
Mansfield John W.,
Grant Murray
Publication year - 2006
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500172
Subject(s) - proteome , biology , arabidopsis thaliana , proteomics , phosphorylation , pseudomonas syringae , phosphoproteomics , protein phosphorylation , arabidopsis , quantitative proteomics , microbiology and biotechnology , signal transduction , heat shock protein , computational biology , chaperone (clinical) , hypersensitive response , kinase , biochemistry , pathogen , protein kinase a , genetics , plant disease resistance , mutant , gene , medicine , pathology
Abstract Despite recent advances in proteomic technologies, quantitative analysis of the proteome remains a challenging task. Phosphorylation of proteins is central to signal transduction pathways and plays an important role in plant defence against pathogens, although the immediate targets of kinases remain elusive. Determining changes in the phosphoproteome during the defence response is a major goal in molecular plant pathology. In this first description of the novel mass tagging strategy (iTRAQ™ Applied Biosystems) applied to plant pathogen interactions, we describe early changes to the phosphoproteome of Arabidopsis thaliana during the defence response to Pseudomonas syringae pv. tomato DC3000. We identified five proteins which showed reproducible differences between a control and three different bacterial challenges, thus identifying proteins potentially phosphorylated as part of a plant basal defence response. Four of the five proteins a dehydrin, a putative p23 co‐chaperone, heat shock protein 81 and a plastid‐associated protein (PAP)/fibrillin, are known to be phosphorylated or have potential phosphorylation sites. One further protein, the large subunit of Rubisco, showed a significant difference between tissue undergoing the hypersensitive response and a basal defence response. We document the reproducibility, utility and problems associated with this approach.