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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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Synthesis of ellipticine and its pharmacologically more efficient derivative 13-hydroxyellipticine
Hájek, Jan ; Černá, Věra (referee) ; Stiborová, Marie (advisor)
Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazole) is plant alcaloid isolated from Ochrosia elliptica1) (Apocyanaceae). Ellipticine's anticancer effect is mainly caused by its intercalation into DNA, formation of covalent adduct with DNA and inhibition of topoisomerase II. Known syntheses of ellipticine are based on chemicals containing two or three aromatic cycles that are used in cyklisation and/or addition reaction to get ellipticine or ellipticine derivates. 13- hydroxyellipticine can be prepared de novo as a product of casual ellipticine synthesis, or as a modification of already synthetised ellipticine. Keywords: ellipticine; synthesis; derivates of ellipticine
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Comprehensive care for a patient with acute necrotizing pancreatitis
Černá, Věra ; Výborný, Josef (advisor) ; Novák, František (referee)
Master's thesis "Comprehensive care of patients with acute necrotizing pancreatitis" is divided into two main parts: theoretical one and a practical one. The theoretical part deals the characteristics of an acute pancreatitis, pancreas history knowledge, aetiology, pathophysiology, classification, clinical picture, diagnosis, therapy and possible complications of acute pancreatitis. In the practical part there are described two cases of the acute necrotizing pancreatitis disease. The first case is solved by the conservative way and the second one by a surgical method.
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Manufacturing of pin with flange
Černá, Věra ; Štroner, Marek (referee) ; Podaný, Kamil (advisor)
The project elaborated within the Bachelor study of the branch Manufacturing technology presents a technology proposal of a forged piece - pin with a flange -madeby drop forging from material 15 142. The forged piece will be carried out by several hits in the finishing die. Regarding to the seriality, 800 000 pieces per year, and calculations of necessery strength for the last hit, the forged piece will be made on a counterblow hammer KHZ-2 with energy of 20 kJ. Forging dies are made of tool steel 19552and heat treated according to the drawing docummentation.
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Manufacturing of pin with flange
Černá, Věra ; Podaný, Kamil (advisor)
The project elaborated within the Bachelor study of the branch Manufacturing technology presents a technology proposal of a forged piece - pin with a flange -madeby drop forging from material 15 142. The forged piece will be carried out by several hits in the finishing die. Regarding to the seriality, 800 000 pieces per year, and calculations of necessery strength for the last hit, the forged piece will be made on a counterblow hammer KHZ-2 with energy of 20 kJ. Forging dies are made of tool steel 19552and heat treated according to the drawing docummentation.
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