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Phosphoproteomics Profiling of Tobacco Mature Pollen and Pollen Activated in vitro
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SYSNO ASEP 0461234 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Phosphoproteomics Profiling of Tobacco Mature Pollen and Pollen Activated in vitro Author(s) Fíla, Jan (UEB-Q) RID, ORCID
Radau, S. (DE)
Matros, A. (DE)
Hartmann, A. (DE)
Scholz, U. (DE)
Feciková, Jana (UEB-Q) ORCID
Mock, H. P. (DE)
Čapková, Věra (UEB-Q) RID
Zahedi, R. P. (DE)
Honys, David (UEB-Q) RID, ORCIDSource Title Molecular & Cellular Proteomics. - : Elsevier - ISSN 1535-9476
Roč. 15, č. 4 (2016), s. 1338-1350Number of pages 13 s. Language eng - English Country US - United States Keywords OXIDE AFFINITY-CHROMATOGRAPHY ; ARABIDOPSIS-THALIANA ; PROTEIN-TYROSINE PHOSPHORYLATION Subject RIV EB - Genetics ; Molecular Biology R&D Projects GA15-16050S GA ČR - Czech Science Foundation (CSF) GAP305/12/2611 GA ČR - Czech Science Foundation (CSF) LD13049 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA15-22720S GA ČR - Czech Science Foundation (CSF) Institutional support UEB-Q - RVO:61389030 UT WOS 000373992600012 DOI 10.1074/mcp.M115.051672 Annotation Tobacco mature pollen has extremely desiccated cytoplasm, and is metabolically quiescent. Upon re-hydration it becomes metabolically active and that results in later emergence of rapidly growing pollen tube. These changes in cytoplasm hydration and metabolic activity are accompanied by protein phosphorylation. In this study, we subjected mature pollen, 5-min-activated pollen, and 30-min-activated pollen to TCA/acetone protein extraction, trypsin digestion and phosphopeptide enrichment by titanium dioxide. The enriched fraction was subjected to nLC-MS/MS. We identified 471 phosphopeptides that carried 432 phosphorylation sites, position of which was exactly matched by mass spectrometry. These 471 phosphopeptides were assigned to 301 phosphoproteins, because some proteins carried more phosphorylation sites. Of the 13 functional groups, the majority of proteins were put into these categories: transcription, protein synthesis, protein destination and storage, and signal transduction. Many proteins were of unknown function, reflecting the fact that male gametophyte contains many specific proteins that have not been fully functionally annotated. The quantitative data highlighted the dynamics of protein phosphorylation during pollen activation; the identified phosphopeptides were divided into seven groups based on the regulatory trends. The major group comprised mature pollen-specific phosphopeptides that were dephosphorylated during pollen activation. Several phosphopeptides representing the same phosphoprotein had different regulation, which pinpointed the complexity of protein phosphorylation and its clear functional context. Collectively, we showed the first phosphoproteomics data on activated pollen where the position of phosphorylation sites was clearly demonstrated and regulatory kinetics was resolved. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2017
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