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pmic7231-sup-0001-FigureS1.tif613KFigure S1. Comparison of the phosphorylation in vitro and 2D-GE spectra of the phosphoprotein-enriched eluate. (A) 11.25% polyacrylamide resolving gel, radiography, (B) 10% polyacrylamide gel, pI 4–7, RuBP staining. The numbers on the left indicate molecular weight in kDa. The letters group the probably corresponding spots and bands together.
pmic7231-sup-0002-FigureS2.pdf1762KFigure S2. MS/MS spectra for the peptides shown in Table 3.
pmic7231-sup-0003-TableS1.xls47KTable S1. Identified phosphoprotein candidates from the excised 2D-GE spots analyzed as described in “Materials and methods” section with additional data (identified peptides, their m/z and ladder scores). Proteins that were identified from at least two gels according to at least two peptides are marked with two asterisks behind the spot number, whereas proteins that were identified only once according to at least two peptides are marked with one asterisk. Superscript behind accession number indicates database: a) Swissprot; b) Trembl Viridiplantae; c) TIGR EST Solanaceae. The superscripted S behind the MP/PT ratio means a significant change between the two stages as calculated by the PDQuest software. Note: Spot 4 was not recognized as a spot by the PDQuest software, but manually assigned
pmic7231-sup-0004-TableS2.xls336KTable S2. Complete set of identified phosphoprotein candidates by the nongel approach analyzed as described in “Materials and methods” section with additional data (identified peptides, their mass, charge, scores and individual quantity data). Number of peptides indicates the total number of peptides, according to which the protein was identified. The number in the brackets represents number of unique peptides that contribute to protein identification (the peptides common for more proteins were excluded). The data regarding diagnostic peptides are shown in black whereas the conflicting ones are displayed in gray
pmic7231-sup-0005-TableS3.xls44KTable S3. List of phosphorylation sites identified after the phosphopeptide enrichment from mature pollen crude extract performed by titanium dioxide chromatography with additional information. The proteins are grouped according to their names, in some cases listing together more isoforms of the same protein since they are hard to be distinguished between each other due to tobacco being not completely sequenced. The phosphorylation site is highlighted in bold and the given phosphorylated amino acid in followed by an asterisk. The ambiguously identified phosphopeptides are shown in gray. The superscripted number behind the accession number indicates database: a) UniProt, 90% homology clusters, Viridiplantae, b) TIGR EST Nicotiana tabacum. MH+ [Da] – molecular weight in daltons. pRS Sc. – pRS Score. Mascot Ion Sc. – Mascot Ion Score. # MC – number of mis-cleavages
pmic7231-sup-0006-S1.doc28KSupporting Information

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