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Structural characterization of influenza A polymerase PA subunit domains in complex with novel inhibitors
Radilová, Kateřina
Influenza RNA-dependent RNA polymerase is a heterotrimeric complex and has an essential role in the life cycle of the virus. It is responsible for viral replication and transcription. One of its subunits, the polymerase acidic protein, interacts with the PB1 subunit via a crucial protein- protein interaction at its C-terminal domain. This 310 helix-mediated intersubunit interaction is required for the whole heterotrimer assembly. The N-terminal domain carries the endonuclease active site with two manganese ions. Both domains are considered promising drug targets. Current strategies to fight the influenza virus are limited to seasonal vaccines, and there are only a few anti-influenza drugs targeting mostly other viral proteins. Many used antivirals are susceptible to rapid resistance mutations development or cause severe side effects. This thesis provides structural insights into the two domains of the PA subunit. The first part is devoted to the characterization and optimization of a PB1-derived minimal peptide interacting with the C-terminal domain. Results from this part may be considered as a starting point for the rational design of first-in-class anti-influenza inhibitors of the PA-PB1 protein-protein interaction. In the other half, we have explored the inhibitory potency of flavonoids and...
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Structural characterization of influenza A polymerase PA subunit domains in complex with novel inhibitors
Radilová, Kateřina ; Kožíšek, Milan (advisor) ; Rumlová, Michaela (referee) ; Obšil, Tomáš (referee)
Influenza RNA-dependent RNA polymerase is a heterotrimeric complex and has an essential role in the life cycle of the virus. It is responsible for viral replication and transcription. One of its subunits, the polymerase acidic protein, interacts with the PB1 subunit via a crucial protein- protein interaction at its C-terminal domain. This 310 helix-mediated intersubunit interaction is required for the whole heterotrimer assembly. The N-terminal domain carries the endonuclease active site with two manganese ions. Both domains are considered promising drug targets. Current strategies to fight the influenza virus are limited to seasonal vaccines, and there are only a few anti-influenza drugs targeting mostly other viral proteins. Many used antivirals are susceptible to rapid resistance mutations development or cause severe side effects. This thesis provides structural insights into the two domains of the PA subunit. The first part is devoted to the characterization and optimization of a PB1-derived minimal peptide interacting with the C-terminal domain. Results from this part may be considered as a starting point for the rational design of first-in-class anti-influenza inhibitors of the PA-PB1 protein-protein interaction. In the other half, we have explored the inhibitory potency of flavonoids and...
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Gene expression of specific parts of CNS in a rat model for transgenerational transmission of drug addiction.
Radilová, Kateřina ; Černá, Marie (advisor) ; Rothová, Olga (referee)
Methamphetamine (MA) is a drug often abused by drug-addicted women and also is the mostly used drug in the Czech Republic. This drug crosses the placental and hematoencephalic barriers easily and therefore it can negatively affect not just mother herself, but also the development of the fetus. Even though there have been several published studies evincive negative effects of MA to posterity of drug-addicted mothers, the most critical term during pregnancy for the fetus remains unknown. In this study are examined CNS structures, which are directly influenced by methamphetamine action. Their function may lead to drug addiction through pleasurable feelings, such as reward feeling evoking. In the next chapter are analysed neurotransmitters and their transporters, which are related to methamphetamine effects. Studying their structure, localization or development during life may help to bring near the drug functioning in different life periods. Methamphetamine itself and its effects are closer described in the last part of this study. The problem of methamphetamine exposed fetus is outlined there - prenatal and postnatal so far known differences, but also sex differences in methamphetamine pharma- cokinetics.
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The analysis of structural details of the NMDA receptor
Radilová, Kateřina ; Balík, Aleš (advisor) ; Jakubík, Jan (referee)
NMDA receptor is necessary for excitatory transmission in the central nervous system. Altered funtion of the NMDA receptors is associated with many neurodegenerative and neuropsychiatric diseases. All available crystal structures of the NMDAR meant great shift towards our understanding of details of the receptor and its function. Unfortunately, these up- to-date available structures present only certain functional states of receptors and also a few structural data are still missing. For complete comprehension of the process of activation and deactivation of NMDA receptors, we need to supplement the current information with more data. The aim of this thesis was to employ a combination of different approaches (computational modelling, cloning, biochemistry, protein expression and purification and mass spectrometry) to obtain new structural data, by which we would be able to fill in the gaps in current receptor models, especially at various functional states of the receptor. Key words: NMDA receptor, glutamate receptor, computational modelling, structure, cloning, protein expression
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Gene expression of specific parts of CNS in a rat model for transgenerational transmission of drug addiction.
Radilová, Kateřina ; Černá, Marie (advisor) ; Rothová, Olga (referee)
Methamphetamine (MA) is a drug often abused by drug-addicted women and also is the mostly used drug in the Czech Republic. This drug crosses the placental and hematoencephalic barriers easily and therefore it can negatively affect not just mother herself, but also the development of the fetus. Even though there have been several published studies evincive negative effects of MA to posterity of drug-addicted mothers, the most critical term during pregnancy for the fetus remains unknown. In this study are examined CNS structures, which are directly influenced by methamphetamine action. Their function may lead to drug addiction through pleasurable feelings, such as reward feeling evoking. In the next chapter are analysed neurotransmitters and their transporters, which are related to methamphetamine effects. Studying their structure, localization or development during life may help to bring near the drug functioning in different life periods. Methamphetamine itself and its effects are closer described in the last part of this study. The problem of methamphetamine exposed fetus is outlined there - prenatal and postnatal so far known differences, but also sex differences in methamphetamine pharma- cokinetics.
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