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Proteomická a bioinformatická charakterizace N-terminálních sekvencí proteinů modifikovaných po importu do hydrogenosomu Trichomonas vaginalis.
Zákoucká, Eva ; Man, Petr (advisor) ; Šulc, Miroslav (referee)
Trichomonas vaginalis is a human pathogen causing trichomoniasis, one of the most common non-viral sexually transmitted diseases in both men and women. Trichomoniasis is currently treated with metronidazole, but the pathogen is known to develop resistance against this drug. However as the pathogen is eukaryotic, the targets for the pathogen elimination without seriously affecting the host are limited. Throughout the evolution Trichomonas vaginalis adapted to anaerobic environments by developing an alternative metabolism resulting in a reduced form of mitochondria named hydrogenosome. Hydrogenosomes lack genetic information, therefore all its proteins are nucleus-encoded and need to be transported inside the hydrogenosome using a targeting N-terminal presequence. The peptidase recognizing and cleaving those presequences at the entrance of the organelle, the hydrogenosomal processing peptidase (HPP), is unique for hydrogenosomes and therefore represents a potential drug target against the pathogen. In this work the HPP's substrate specificity towards the targeting presequences was investigated. To do so a proteomic analysis of the proteome of Trichomonas vaginalis hydrogenosomes was performed using a novel optimized protocol for N-terminal peptide sequencing. N-terminal peptides were captured using a...
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Proteomická a bioinformatická charakterizace N-terminálních sekvencí proteinů modifikovaných po importu do hydrogenosomu Trichomonas vaginalis.
Zákoucká, Eva ; Man, Petr (advisor) ; Šulc, Miroslav (referee)
Trichomonas vaginalis is a human pathogen causing trichomoniasis, one of the most common non-viral sexually transmitted diseases in both men and women. Trichomoniasis is currently treated with metronidazole, but the pathogen is known to develop resistance against this drug. However as the pathogen is eukaryotic, the targets for the pathogen elimination without seriously affecting the host are limited. Throughout the evolution Trichomonas vaginalis adapted to anaerobic environments by developing an alternative metabolism resulting in a reduced form of mitochondria named hydrogenosome. Hydrogenosomes lack genetic information, therefore all its proteins are nucleus-encoded and need to be transported inside the hydrogenosome using a targeting N-terminal presequence. The peptidase recognizing and cleaving those presequences at the entrance of the organelle, the hydrogenosomal processing peptidase (HPP), is unique for hydrogenosomes and therefore represents a potential drug target against the pathogen. In this work the HPP's substrate specificity towards the targeting presequences was investigated. To do so a proteomic analysis of the proteome of Trichomonas vaginalis hydrogenosomes was performed using a novel optimized protocol for N-terminal peptide sequencing. N-terminal peptides were captured using a...
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Inhibition of Thymidine Phosphorylase
Zákoucká, Eva ; Brynda, Jiří (advisor) ; Obšil, Tomáš (referee)
2. Abstract Thymidine phosphorylase (TPase), also known as gliostatin or Platelet-derived endothelial cell growth factor (PD-ECGF), is an enzyme with an important role in the nucleoside metabolism and is also involved in degradation and recycling of DNA. TPase catalyzes the reversible phosphorolysis of pyrimidine 2'-deoxynucleosides to 2-deoxy-D-ribose-1- phosphate and their respective bases, as well as the transfer of the deoxyribosyl moiety from one pyrimidine base to another. Thymidine phosphorylase is a therapeutic target of great importance because of its participation in angiogenesis especially in solid tumors of various tissues. Therefore, TPase stimulates tumor growth and progression, as well as metastasis. In addition to this, TPase inhibits apoptosis, particularly of tumor cells and causes degradation of several antiviral and anticancer drugs. Apart from the carcinoma tissues, thymidine phosphorylase is overexpressed in various other tissues affected by disorders characterized by proliferation of blood vessels including psoriasis, rheumatoid arthritis and atherosclerosis. Inhibiting the activity of TPase selectively in the tissues affected by the diseases listed above would be of great therapeutic significance. Therefore, many inhibitors, mainly substrate analogues, have been designed based on the...
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