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Iron metabolism in Naegleria gruberi
Arbon, Dominik ; Šuťák, Róbert (advisor) ; Krijt, Matyáš (referee)
The metabolism of iron ions is a crucial process in all living organisms and its correct regulation is essential for basic life functions. Homeostasis of iron ions is closely regulated, it usually appears as a component of various proteins and plays role in many oxidation-reduction reactions. Naegleria gruberi is a non-pathogenic, free living protozoon, that serves as a laboratory model for closely related pathogenic Naegleria fowleri. This work focuses on the study of selected metabolites of N. gruberi, that were possible to detect and quantify by the means of modern metabolomic methods, and the influence on culture cultivated in environment with lack of iron ions was shown. The discovery of effect of this condition on the energetic metabolism of this protozoan is an important aspect of understanding the biological processes on cellular level. This method proved a significant influence on certain metabolites and modification of certain metabolic pathways as a direct effect of decreased availability of iron ions. Second part of this work was focused on the enzyme alcohol dehydrogenase, that was found in the genome of this protozoon. Unusual aspects of this enzyme include a N-terminal mitochondrial presequence, prompting about mitochondrial localization, and utilization of iron ion as a prosthetic...
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The effect of iron on the metabolism of the protozoan Trypanosoma brucei
Arbon, Dominik ; Konvalinka, Jan (advisor) ; Grantz Šašková, Klára (referee)
Iron ions are essential components of numerous cell processes. Their utilisation is strictly regulated, since any impairment can have devastating effect on the cell. In living organisms, iron ions are bound to proteins, for storage, transportation, or as a vital part of catalytic centers of enzymes. Transportation of iron ions between different compartments is important for the correct function of the cell. It was recently shown on yeast, how a mitochondrial transporter of iron ions is essential for the synthesis of iron-sulfur clusters of enzymes. This work aims to describe the localization and function of a homologous protein of a parasitical organism Trypanosoma brucei, which causes African trypanosomiasis, also known as sleeping sickness. This parasite is entirely dependent on uptake of iron ions from its host and therefore the utilization of iron ions is studied as a potential therapeutic target. This work is focused on the characterization of protein Mcp17, which is assumed to function as a transporter of iron ions into the mitochondria of T. brucei. Utilizing expression of marked Mcp17, the transporter was confirmed to be localized on the mitochondrial membrane of the cell. Measuring of enzyme activity of selected enzymes indicated that cells with inhibited expression of the gene mcp17...
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