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Quantum dot luminescent probe for Caspase 3/7 imaging inside cells
Procházková, Markéta ; Klepárník, Karel
Two step synthesis of luminescent probe based on ligand-exchange in the first step and the specific reaction of amino group in the second step was optimized. The luminescence properties of the final product were checked by time course of\nactive recombinant caspase protein reaction under the model conditions in fluorimeter. The novel luminescent probe enables long-time imaging of active caspases in living cells or tissues.
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Role of DD- and DED-containing adaptor proteins in apoptotic signaling
Čaja, Fabián ; Anděra, Ladislav (advisor) ; Janštová, Vanda (referee)
Proteins containing a bundle of six anti-paralel α-helices in so-called "death domain" (DD) and similar structures (DED, CARD) represent important players in apoptotic signaling. To DD/DED/CARD domains-containing proteins belong pro- apoptotic membrane receptors from the TNFR superfamily, then adaptor proteins and enzymes as proteases or kinases. These pro-apoptotic "death receptors" interact with adaptor proteins and initiator caspases containing DDs or DEDs and activate apoptotic signaling cascade. DEDs and DDs are in addition found in many proteins participating in activation of caspases or other non-apoptotic signaling. Many experimental models document that defects in and deregulations of proteins containing DDs and DEDs can have severe if not lethal consequences for an organism. Abberations in these proteins in many cases could lead to cancerogenesis, immunodeficiencies or developmental defects.
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Molecular mechanism of MST1 kinase activation in cancer cells
Smetanová, Jitka ; Vališ, Karel (advisor) ; Grobárová, Valéria (referee)
MST1 kinase is an internal part of the Hippo signal pathway. The Hippo pathway is an evolutionary conserved regulator of tissue and organ growth and affects proliferation and apoptosis. Active MST1 kinase phosphorylates YAP and TAZ oncoproteins, which regulate the activity of transcription factors in their unphosphorylated state, including TEAD and SMAD. Furthermore active MST1 kinase phosphorylates FOXO transcription factors and induces their translocation into the cell nucleus. Finally the activation of MST1 kinase leads to cell apoptosis or halt cell cycle in G1 phase. Activation of MST1 protein depends on its auto-phosphorylation and cleavage. Recently, there are several articles which take interest in the issue of activation of MST1. Some of them describe the activation of MST1 by the effector caspase-3 and -7, on the other hand the latest articles argue that MST1 kinase itself is responsible for the activation of caspases. The molecular mechanism of MST1 kinase activation was studied in this bachelor thesis. We used the biologically active compounds GDC-0941 for the activation of MST1 protein. The activity of caspase was inhibited by specific inhibitor Z-DEVD. Using electrophoresis and Western blot it was demonstrated that MST1 is active in the case when caspases are inhibited. This fact...
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Histone deacetylase inhibitors induced caspase-independent cell death
Groh, Tomáš ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
Neuroblastoma is the most common extracranial solid tumor that occurs during infancy. Despite the great progress has been made in contemporary clinic medicine some forms of neuroblastoma disease are still found very difficult to treat . This work focuses on the effects of histone deacetylase inhibitors (HDAC) in the neuroblastoma cell lines. It is known that HDAC inhibitors may contribute to recurrence of the tumor cells by affecting the chromatin structure and thus increase the expression of critical tumor suppressor genes. These genes activate apoptotic pathways that may even be independent of caspases. We observed the efficiency of used HDAC inhibitors as under standard conditions an in hypoxia (1 % O2). Inadequate amount of oxygen supply is one of the characteristic features of tumors and it also may contribute to chemoresistance. With the hypoxia-induced chemoresistance of tumor cells, the influence of HIF-1α is expected. Some HDAC inhibitors reduce the amount of HIF-1α in hypoxia and thus HIF transcription factor activity. Thus, the first part of this study is concerned with the acquisition of suitable experimental arrangement for the monitoring of induction of cellular death in human neuroblastoma cell lines SK-N-AS and UKF-NB-3. Secondly, this paper provides the evaluation of the influence...
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Role of DD- and DED-containing adaptor proteins in apoptotic signaling
Čaja, Fabián ; Janštová, Vanda (referee) ; Anděra, Ladislav (advisor)
Proteins containing a bundle of six anti-paralel α-helices in so-called "death domain" (DD) and similar structures (DED, CARD) represent important players in apoptotic signaling. To DD/DED/CARD domains-containing proteins belong pro- apoptotic membrane receptors from the TNFR superfamily, then adaptor proteins and enzymes as proteases or kinases. These pro-apoptotic "death receptors" interact with adaptor proteins and initiator caspases containing DDs or DEDs and activate apoptotic signaling cascade. DEDs and DDs are in addition found in many proteins participating in activation of caspases or other non-apoptotic signaling. Many experimental models document that defects in and deregulations of proteins containing DDs and DEDs can have severe if not lethal consequences for an organism. Abberations in these proteins in many cases could lead to cancerogenesis, immunodeficiencies or developmental defects.
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