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Protein-protein interaction mapping of cytochrome P-450 by methods using chemical modification and mass spectrometry
Ječmen, Tomáš ; Černá, Věra (referee) ; Šulc, Miroslav (advisor)
Cytochromes P-450 (P450s) belong to haemoprotein superfamily and they are responsible for metabolism of a wide variety of compounds, among others many drugs and carcinogens. P450s serve as the terminal oxidases in the mixed function oxidase system in cooperation with a redox partner NADPH: cytochrome P450 reductase (CPR) providing input of two electrons to the reaction cycle of P450. The CPR can be substituted by other redox partner of P450, cytochrome b5 (cyt b5), to deliver the second electron. Three dimensional structure of P450 is required in order to fully understand its reaction mechanism. At the present time, a homology model of cytochrome P-450 2B4 (CYP 2B4) is available in our laboratory. In this study, the mapping of interaction domain between CYP 2B4 and cyt b5 employing a crosslinking agent EDC to form amide bonds between close complementary charged amino acid side chains was the first goal. We have identified five interacting amino acid pairs in total using mass spectrometry (MS). The second research interest was to verify and refine the CYP 2B4 model using a photoaffinity labelling with N-(p-azidobenzyl)-N-methyl-p-aminophenylamine probe. This photoreactive probe is known as CYP 2B4 ligand binding to the central iron atom of haem. After photoactivation the arginine 197 was found by MS...
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Study of the microRNAs in patients with head and neck squamous cell carcinoma
Hudcová, Kristýna ; Šulc, Miroslav (referee)
Searching for markers of disease is at the forefront of scientific interest. Biomarker of a specific disease should help in disease management - whether for diagnosing a disease or predicting its progress. We picked microRNAs - short RNAs that regulate mRNA translation, due to their stability and function in the regulation of many cellular processes. Therefore they can be used as a biomarker for many diseases. In our study we focused on neck squamous cell carcinoma (HNSCC), since this type of cancer is widespread and clinically important markers are missing. It is in our interest to seek new kinds of markers, suitable for early diagnosis of the disease and the most accurate assessment of prognosis. We selected 3 candidate miRNA (miR-29c-3p, miR-200b-5p, miR-375-3p) and used qRT-PCR method to profile their expression in tissues of patients with HNSCC. MiRNA expressions in 42 tumor samples were compared with the miRNA expression in adjacent tissue of the tumor. Adjacent tissue surrounds the tumor and has interesting properties, as it looks like a histological normal tissue, but on closer inspection it is not. It is affected by tumor tissue and may contribute to tumor progression. We managed to distinguish between these two tissues using miR-29c-3p and miR-375-3p. Additional analyzes were focused on...
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Planning and evalauation of corporate strategy in online social media environment
Šulc, Miroslav ; Karlíček, Miroslav (advisor) ; Kolář, Petr (referee)
This paper researches the problem of creation an efficient strategy for the environment of online social networks, and its' evaluation. Theoretical part of the paper progresses gradually from definition of social media, its' progress into online world, brief description of historical development and finally covers strategic marketing approaches specific for this field. The practical part of the paper contains few case studies and my own social media campaign I ran for e-shop JustHappy.cz. The conclusion is a synthesis of researched ideas and proposition of step-by-step manual for preparing social media strategy. The manual is broken down to several easy-to-follow steps and should give the reader a handbook for creating his/her own campaign or project in online social media environment.
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Panel of monoclonal antibodies – alternative tool For monitoring of sperm–zona pellucida receptors localization and identification
Zigo, Michal ; Dorosh, Andriy ; Pohlová, Alžběta ; Jonáková, Věra ; Šulc, Miroslav ; Maňásková-Postlerová, Pavla
Primary binding of the sperm to the zona pellucida (ZP) is one of the many steps necessary for successful fertilization in all sexually reproducing species. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-precised sequential process. Primary-binding receptors are localized throughout the acrosomal region of the sperm surface of which many have been disclosed in various mammals. For the monitoring sperm-zona pellucida receptors in terms of localization and characterization - panel of monoclonal antibodies against proteins from the sperm surface was prepared. Antibodies were screened by immunofluorescence and Western blotting for protein localizations and competence of antibodies, respectively. Antibodies recognizing proteins localized on the sperm head and simultaneously detected by Western blot were further studied by means of immunolocalization in reproductive tissues and fluids, binding to ZP, immunoprecipitation and protein identification using MS analysis. Out of 17 prepared antibodies, 8 antibodies were simultaneously recognizing proteins localized on the sperm head and detecting proteins of interest by Western blotting. Further only 3 antibodies recognized proteins which also coincided in binding to ZP. These 3 antibodies were used for immunoprecipitation, and further protein identification of immunoprecipitates revealed that the antibodies distinguish acrosin precursor, RAB2A protein, and lactadherin P47. Acrosin and lactadherin P47 have been already detected on the sperm surface and their physiological functions in reproduction have been proposed. To our knowledge, this is the first time RAB2A has been found on the surface of sperm and its physiological function in the process of fertilization remains undisclosed.
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Theory of behavioral finance
Šulc, Miroslav ; Kuncl, Martin (advisor)
The bachelor thesis is dedicated to behavioral finance and it's application. The thesis describes evolution of the theory till present. Theories are introduced, which forego behavioral finance, for example Expected utility theory. Just as well the newest discipline, neuroeconomics, is outlined. The application part of the thesis analyse bubble on technological market in USA 1995-2000.
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