title: Phosphorylation site mapping of soluble proteins: bioinformatical filtering reveals potential plastidic phosphoproteins in Arabidopsis thaliana.
abstract: Protein phosphorylation is a major mode of regulation of metabolism, gene expression, and cell architecture. A combination of phosphopeptide enrichment strategies based on TiO(2) and IMAC in addition to our MudPIT strategy revealed the detection of 181 phosphorylation sites which are located on 125 potentially plastidic proteins predicted by GoMiner, TargetP/Predotar in Arabidopsis thaliana. In our study phosphorylation on serine is favored over threonine and this in turn over phosphorylation on tyrosine residues, showing a percentage of 67.4% to 24.3% to 8.3% for pS:pT:pY. Four phosphorylated residues (S208, Y239, T246 and T330), identified by our approach have been fitted to the structure of the activated form of spinach RuBisCO, which are located in close proximity to the substrate binding site for ribulosebisphosphate. Potentially, these phosphorylation sites exert a direct influence on the catalytic activity of the enzyme. Such examples show nicely the value of the presented mass spectrometric dataset for further biochemical applications, since alternative mutation analysis often turns out to be unsuccessful, caused by mutations in essential proteins which result in lethal phenotypes.
authors.affiliation: Department of Analytical Chemistry, Ruhr-University Bochum, Bochum, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Lohrig
authors.forename: Bernd
authors.initials: B
authors.lastname: Müller
authors.forename: Joulia
authors.initials: J
authors.lastname: Davydova
authors.forename: Dario
authors.initials: D
authors.lastname: Leister
authors.forename: Dirk Andreas
authors.initials: DA
authors.lastname: Wolters
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000032:analysis*; D019295:Computational Biology / Q000379:methods*; D010750:Phosphoproteins / Q000032:analysis*; D010766:Phosphorylation; D018087:Plastids / Q000378:metabolism*; D015203:Reproducibility of Results; D012984:Software; D012995:Solubility
chemical_list: D029681:Arabidopsis Proteins; D010750:Phosphoproteins