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National Repository of Grey Literature 2 records found  Search took 0.00 seconds. 
Studium ubiquitinace proteinů DELLA
 Breineková, Alžběta
Posttranslational modifications (PTMs) of proteins represent fascinating extensions of the dynamic complexity of living cells' proteomes, but also present a solid obstacle in the proteome analysis. Identification and mapping of PTMs in proteins have improved dramatically, but to comprehend complex mechanisms and biological functions, one must address also a very low abundant proteins. Here, in this thesis entitled 'Analysis of DELLA protein ubiquitination' we demonstrate the use of a recombinant protein standard for the determination of in vivo modified peptides of the DELLA family protein RGA. The candidate peptide sequences were targeted in an SRM-based analysis to detect the ubiquitination site and the results of this analysis and that of an MS/MS data processing indicate that the modification is localized in the conserved N-terminal region of RGA protein.
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Metody frakcionace a obohacování rostlinného proteomu
 Breineková, Alžběta
The proteome is a complex and dynamic mixture of proteins produced by an organism. It represents tens of thousands different protein species and its characterization is a difficult task even with state-of-the-art techniques. This thesis entitled "Methods for plant proteome fractionation and enrichment" is focused on plant proteomics and approaches that can improve detection limits and increase proteome coverage. In the experimental part, model plant Arabidopsis thaliana proteome was analysed. The tested techniques include crude fractionations on the tissue level, a subcellular fractionation, and separations on the protein level. Finally, sets of samples were analysed via LC-MS and the method for peptide analysis was optimized. Combining the LC-MS datasets revealed more than 4900 identified Arabidopsis proteins (FDR<1%). Of these, 248 were detected only in equalized proteome. The results of this work helped to establish a robust proteome analysis workflow that will be implemented in plant proteome research at MENDELU.
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