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Production of recombinant cathepsin C from human blood fluke
Illichová, Hana ; Konvalinka, Jan (advisor) ; Martínková, Markéta (referee)
Blood flukes of the genus Schistosoma cause schistosomiasis, a serious parasitic disease occurring in tropical and subtropical areas. Cathepsin C (EC 3.4.14.1) is a digestive enzyme of the blood flukes which participates in the degradation of hemoglobin through its dipeptidyl aminopeptidase activity. This enzyme is critical for metabolism of the parasite and represents a potential target for the development of antischistosomal drugs. Cathepsin C has not yet been studied in detail. This bachelor thesis is focused on the development of expression systems for production of recombinant cathepsin C of Schistosoma mansoni (SmCC). The yeast Pichia pastoris system was used for the expression of an inactive SmCC precursor (zymogen) whose proteolytic stability was analysed. Furthermore, the expression system for SmCC in the protozoan Leishmania tarentolae was employed, and four different SmCC constructs were prepared to optimize production.
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Mitochondrial proteases and their role in biogenesis of mitochondria
Krump, Ondřej ; Stibůrek, Lukáš (advisor) ; Pecina, Petr (referee)
Mitochondria are organelles of endosymbiotic origin responsible for many cellular functions, including bioenergetics, biosynthesis and apoptosis. Regulated protein turnover is crucial for proper mitochondrial function. It is controlled by cellular proteolytic system, especially by its mitochondrial part. This mitochondrial proteolytic system is comprised of severeal groups of proteases. The best characterized AAA+ proteases constitute hollow oligomeric complexes, in which the proteolytic domains are localized. Access to these domains is dependent on unfolding - an energy-consuming process driven by ATP hydrolysis mediated by ATPase domains of AAA+ protein. The main function of AAA proteases is proteolytic degradation of proteins, a part of quality control system of mitochondrial proteins. AAA proteases are localized freely in mitochondrial matrix (Lon and ClpXP), or anchorred in the inner membrane (i-AAA and m-AA). Processing peptidases cleave off the mitochondrial targetting sequences of nuclearly encoded mitochondrial proteins. Oligopeptidases cleave peptides produced by processing and proteolytical degradation to single amino acids. Incorrect function of various components of mitochondrial proteolytical system is implicated in several diseases, including certain forms of hereditary spastic paraplegia...
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Cathepsin L by parasites - occurrence, features, functions
Perháčová, Terézia ; Mikeš, Libor (advisor) ; Kašný, Martin (referee)
Cathepsines L are lysosomal cysteine endopeptidases with an universal function in protein catabolism. This work discusses present knowledge about their characteristics in the context of their specific function in parasites. Features and function differences are described in detail on molecular level. The emphasis is on the biochemical properties with resultant use of these enzymes. Cathepsines L of kinetoplastida, aplikomplexa, entamoeba and helmints (focused on Fasciola spp and Schistosoma spp) are each discussed in appropriate chapters. Key words: hydrolase, protease, cysteine peptidase, cathepsin L, lysosome, parasite
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Cathepsin C of the blood fluke Schistosoma mansoni
Oupicová, Irena ; Konvalinka, Jan (advisor) ; Ryšlavá, Helena (referee)
Blood flukes of the genus Schistosoma cause schistosomiasis, one of the most serious parasitic diseases. Cathepsin C (EC 3.4.14.1) is a digestive enzyme of the blood flukes and a potential drug target. This cysteine protease has not been studied in detail yet. In this thesis, cathepsin C activity was detected in the extract of adult blood flukes S. mansoni and S. japonicum. Inhibitory specificity of these enzymes was determined using of set of selective inhibitors of mammalian cathepsins. Cathepsins C were visualized on SDS-PAGE gels with fluorescent proteomic probe. Furthermore, recombinant cathepsin C of S. mansoni (SmCC) was prepared by homologous expression in the yeast Pichia pastoris. (In Czech)
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The study of enzyme activities in compost
Pernicová, Iva ; doc.Mgr.Pavlína Pelcová, Ph.D. (referee) ; Voběrková, Stanislava (advisor)
This thesis deals with the determination of enzyme activities in compost for consideration their changes during the composting. The changes in the activity of particular enzymes serve as the indicators of the activity of microorganisms, which are found in the compost. In the practical part of the work the changes in activity of proteases, cellulases, lipases, dehydrogenases and ureases within 21 days of composting under laboratory conditions were determined. The change in pH was observed as well. The changes in the pH-values and all enzyme activities except ureases present in the first week the same trend associated with the use of available substrates. The changes in the activities of the key enzymes in compost under laboratory conditions were compared with the changes in activities in compost under real conditions, which is the indicator of the different composting process under the given circumstances.
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