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New types of magnetic sorbents for phosphoprotein analysis
Emmerová, Tereza ; Kučerová, Zdenka (advisor) ; Ryšlavá, Helena (referee)
The method for the study of protein phoshorylation sites was elaborated. This method is based on the IMAC separation of phosphopeptides from protein proteolytic digests using new magnetic sorbents and on their subsequent identification by mass spectrometry (MS). Magnetic non-porous hydrophilic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) particles prepared by the dispersion polymerization and modified with iminodiacetic acid (IDA) with immobilized Fe(III) and Ga(III) ions were employed for the enrichment of phosphopeptides from the proteolytic digests of two model proteins, porcine pepsin A and bovine α-casein. The optimum conditions for phosphopeptide adsorption and desorption in both cases were investigated and compared. The phosphopeptides separated from both proteolytic digests were analyzed by matrix-assisted laser desorption/ionization time-of-flight MS. For the immunochemical separation of phosphoproteins, protein fraction containing antibodies was obtained from egg yolk of hens immunized with O-phosphoryl-L-serine conjugated to key limpet hemocyanin. Antibodies were purified using affinity chromatography on immobilized α-casein and their presence was proven by MS. Specificity of the obtained antibodies was examined using ELISA tests. Obtained results showed, that specificity...
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Surviving of Northern Lapwing (Vanellus vanellus) chicks in agriculture landscape
Kubelka, Vojtěch ; Šálek, Miroslav (advisor) ; Cepák, Jaroslav (referee)
The variability of egg-size within a bird species reflects different reproductive investments and, besides the intrinsic constraints and particular female quality, it can be affected by environmental factors, too. Moreover, the egg-size predetermines the chick performance. The knowledge of factors influencing precocial chick survival is crucial for better understanding of life-history strategies as well as for endangered species conservation. The Northern Lapwing (Vanellus vanellus) represents a suitable model species for investigation of climate variables effects on egg-size and also to study factors influencing chick survival. Eggs in a total of 714 nests were measured during 13 breeding seasons over period 1988-2014 in South Bohemia and 293 lapwing chicks from 100 broods in 37 localities were monitored in 2013-2014. The mean egg-size through the whole season is positively affected by March precipitation, March clutches also negatively by minus temperatures in March. The higher intensity of rainfall there is during March, the bigger eggs are laid in March and the steeper is slope of decline in mean egg-size during season. This decline is more profound with lower rainfall in April. The effect of habitat is not significant. The body mass and condition of hatchlings is positively correlated with the...
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Surviving of Northern Lapwing (Vanellus vanellus) chicks in agriculture landscape
Kubelka, Vojtěch ; Šálek, Miroslav (advisor) ; Cepák, Jaroslav (referee)
The variability of egg-size within a bird species reflects different reproductive investments and, besides the intrinsic constraints and particular female quality, it can be affected by environmental factors, too. Moreover, the egg-size predetermines the chick performance. The knowledge of factors influencing precocial chick survival is crucial for better understanding of life-history strategies as well as for endangered species conservation. The Northern Lapwing (Vanellus vanellus) represents a suitable model species for investigation of climate variables effects on egg-size and also to study factors influencing chick survival. Eggs in a total of 714 nests were measured during 13 breeding seasons over period 1988-2014 in South Bohemia and 293 lapwing chicks from 100 broods in 37 localities were monitored in 2013-2014. The mean egg-size through the whole season is positively affected by March precipitation, March clutches also negatively by minus temperatures in March. The higher intensity of rainfall there is during March, the bigger eggs are laid in March and the steeper is slope of decline in mean egg-size during season. This decline is more profound with lower rainfall in April. The effect of habitat is not significant. The body mass and condition of hatchlings is positively correlated with the...
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New types of magnetic sorbents for phosphoprotein analysis
Emmerová, Tereza ; Ryšlavá, Helena (referee) ; Kučerová, Zdenka (advisor)
The method for the study of protein phoshorylation sites was elaborated. This method is based on the IMAC separation of phosphopeptides from protein proteolytic digests using new magnetic sorbents and on their subsequent identification by mass spectrometry (MS). Magnetic non-porous hydrophilic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) particles prepared by the dispersion polymerization and modified with iminodiacetic acid (IDA) with immobilized Fe(III) and Ga(III) ions were employed for the enrichment of phosphopeptides from the proteolytic digests of two model proteins, porcine pepsin A and bovine α-casein. The optimum conditions for phosphopeptide adsorption and desorption in both cases were investigated and compared. The phosphopeptides separated from both proteolytic digests were analyzed by matrix-assisted laser desorption/ionization time-of-flight MS. For the immunochemical separation of phosphoproteins, protein fraction containing antibodies was obtained from egg yolk of hens immunized with O-phosphoryl-L-serine conjugated to key limpet hemocyanin. Antibodies were purified using affinity chromatography on immobilized α-casein and their presence was proven by MS. Specificity of the obtained antibodies was examined using ELISA tests. Obtained results showed, that specificity...
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