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Revealing phosphoproteins playing role in tobacco pollen activated in vitro
Fíla, Jan ; Honys, David (advisor) ; Fischer, Lukáš (referee)
5 Abstract Tobacco mature pollen rehydrates in vivo on a stigma tissue, and develops into the rapidly-growing pollen tube. This rehydration process is accompanied by the de-repression of stored mRNA transcripts, resulting in the synthesis of novel proteins. Furthermore, such metabolic switch is also likely to be regulated on the level of post-translational modifications of the already-present proteins, namely via phosphorylation, since it was shown to play a significant regulatory role in numerous cellular processes. Since only a minor part of proteins is phosphorylated in a cell at a time, the employment of various enrichment techniques is usually of key importance. In this diploma project, metal oxide/hydroxide affinity chromatography (MOAC) with aluminium hydroxide matrix was applied in order to enrich phosphoproteins from the mature pollen and the 30-minute in vitro activated pollen crude protein extracts. The enriched fraction was separated by both 2D-GE and gel-free liquid chromatography (LC) approaches with subsequent mass spectrometric analyses. Collectively, 139 phosphoprotein candidates were identified. Additionally, to broaden the number of phosphorylation sites identified, titanium dioxide phosphopeptide enrichment of trypsin-digested mature pollen crude extract was performed. Thanks to the...
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Enrichment techniques employed in phosphoproteomics
Fíla, Jan ; Honys, David (advisor) ; Zdráhal, Zbyněk (referee)
Reversible protein phosphorylation represents one of the most rapid and dynamic posttranslational modifications. Phosphorylation plays a key role in many cellular processes, e.g. cell division, signal transduction, cytoskeleton dynamics, translation regulation, protein targeting and metabolism. Unfortunately, the importance of phosphoproteins is often not mirrored in their abundance so enriching strategies are mostly inevitable. The enrichment can be performed either at the stage of phosphoproteins or at the stage of phosphopeptides. Alternatively, a combination of both strategies can be used. Most enriching strategies are based on chemical modifications, affinity of phosphates to positively-charged chromatography matrix or antibodies. Every method used has its advantages as well as disadvantages. Moreover, satisfactory coverage of phosphoproteome is very often reached by combination of several methods. In the past five years many protocols have been improved or alternatively new approaches have appeared. So in this bachelor thesis, the available enrichment strategies are reviewed. Key words: phosphoproteomics, phosphoprotein enrichment, phosphopeptide enrichment, IMAC, MOAC, titanium dioxide TiO2, aluminium hydroxide Al(OH)3, antibodies
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Using concertante principle in works by Alfred Schnittke
Fila, Jan ; RIEDLBAUCH, Václav (advisor) ; FILAS, Juraj (referee)
Concertante principle is one of the compositional technic that goes along in the western music
for more than four hundred years. Basis for this historic techniques are used until now. For
researcher arises the question: Exists the concertante principle in the newer musical literature,
or merely is a relic of the distant past? In the last few decades has been one of the strongest
personalities, who dealt with the using of the concertante principle, the Russian-German
composer Alfred Schnittke, who wrote several dozen concertante works. In music history
after the Second World War, we can hardly find any another person that would be dealt with at length this genre. The aim of this thesis is to examine in detail, how the composer treated
with this technique if used only historically given compositional techniques, or innovatively
replaced. As a method of investigation was chosen analysis of large amounts of scores, on
which there is evidence Schnittke use and targeted using of concertante principle.
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Spatial composition
Fila, Jan ; RIEDLBAUCH, Václav (advisor) ; FILAS, Juraj (referee)
This bachelor thesis describes the spatial compositions with a special focus on music scores created since the end of the Second World War to the presence. The aim of thesis is to highlight the artistic value of these works and the particularities of the issue in their performance.
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