|
|
Prolyl endopeptidase of the blood fluke Schistosoma mansoni
Fajtová, Pavla ; Konvalinka, Jan (advisor) ; Ryšlavá, Helena (referee)
Prolyl endopeptidase SmPEP from the blood fluke Schistosoma mansoni is investigated here for the first time. This enzyme is potentially interesting as a drug target for the treatment of schistosomiasis. SmPEP was detected in the extract of adult worms by enzyme activity and immunoreactivity. Enzymatically active SmPEP was produced in the E. coli expression system and was chromatographically purified. The pH optimum of recombinant SmPEP was about 8. Substrate specificity analysis revealed that SmPEP cleaved peptide substrates by endopeptidase activity, however, macromolecular substrates were not fragmented. The residue preferences in the positions P3 to P1' were determined using synthetic fluorogenic peptide substrates. SmPEP was found to be highly sensitive to the inhibition by Z-Ala-Pro-CMK and Z-Arg-Pro-CHO. Primary screening of crystallization conditions for recombinant SmPEP was performed. " (In Czech)"
|
|
|
The role of NADPH oxidase and ros in invadopodia formation
Hanušová, Kristýna ; Vrbacký, Marek (referee) ; Brábek, Jan (advisor)
Invadopodia as specific organelles enabling tumour cells movement, spreading over the organism and ultimately formation of metastasis are possible and promising targets of tumour therapy. Recently, many interesting facts about assembly and mechanism of function of invadopodia were discovered. Invadopodia are centres of ECM degradation by extra-cellular proteases facilitating an invasion of tumour cells. For creation of invadopodia a precisely localized increased production of ROS is necessary. ROS work as crucial signalling molecules and participate in many processes resulting in invadopodia formation. ROS in tumour cells are produced by specific extra-mitochondrial NADPH oxidases (Nox). Several regulatory molecules participating in activation and localization of Nox to invadopodia have been discovered recently (Tks organizer proteins). Furthermore, a regulatory role of Src kinase in ROS production and subsequent invadopodia formation was confirmed. Key words: ECM degradation, invadopodia, invasion, proteases, Nox, ROS, Src kinase, Tks proteins
|
|
|
Role of FtsH proteases in the cyanobacterium Synechocystis sp. PCC 6803
KRYNICKÁ, Vendula
This thesis focuses on the functional and structural characterization of FtsH proteases in Synechocystis PCC 6803. One of the aims was to determine localization and subunit organization of FtsH homologues in Synechocystis cells using GST and GFP tagged FtsH derivatives. The main result of the thesis is identification of two FtsH hetero-oligomeric complexes and one homo-oligomeric complex in Synechocystis cells. The large part of the thesis is aimed at establishing the role of the first hetero-oligomeric complex, FtsH2/FtsH3, in quality control of Photosystem II and at identification of a mechanism, how its substrate proteins D1 and D2 are recognized. Another part is dedicated to characterization of the second hetero-oligomeric complex, FtsH1/FtsH3, which consists of two essential FtsH homologues and which is here identified as an important regulatory element in maintaining iron homeostasis.
|
|
|
Proenzyme therapy of sarcoma S-180 and melanoma B16-F10
KAISEROVÁ, Pavlína
The aim of this study was to evaluate the effectiveness of individual (inactive) proenzymes and mixtures thereof in cancer treatment and to compare this treatment with more frequently used therapy based on active proteases. Experiments focused on explanation of possible mechanisms of proenzyme action against tumors are included.
|
guest ::
