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Analýza slinných serpínů klíšťat Ixodes ricinus z hlediska strukturní biologie
KAŠČÁKOVÁ, Barbora
The knowledge of the detailed structure of proteins and their complexes with other proteins, such as serpins, helps to understand the mechanism of action. Serpins, a large protein group of protease inhibitors that possess almost identical secondary-structural folds, represent the perfect example of expanding the knowledge of their inhibition process through detailed structural analyses. The universal process of serpin inhibition is known, but also is known that serpins are structurally similar whereas their functional diversity is significant. Therefore, each serpin will have some properties specific to its own and knowledge of the serpin structures can explain high functional diversity. X-ray crystallography was one of the most common tools used for serpin structural analysis. This thesis describes the structural information of serpins found in Ixodes ricinus ticks. Serpins of this species are mainly responsible for the modulation of the host immune response via inhibiting involved proteases. Serpins with proteases form covalent complexes. This process leads to a suicide mechanism that inactivates the protease as well as serpin. Here are presented results of the X-ray structural analysis of four I. ricinus serpins named Iripin-3, Iripin-5, Iripin-4, and Iripin-1. All of them help the tick in different ways to stay attached to the host for sufficient time for feeding by inhibiting the proteases involved in host immune defense responses to a tick bite.
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Anti-inflammatory and immunomodulatory activities of Ixodes ricinus salivary proteins
CHLASTÁKOVÁ, Adéla
Ticks are highly specialized obligate ectoparasites whose life cycle is fully dependent on obtaining a blood meal from a host. To prevent blood clotting and counteract the host's immune and wound healing responses, ticks secrete saliva, containing hundreds to thousands of proteins and non-proteins, into the feeding cavity. This thesis focuses on deciphering the function of three serpins (inhibitors of serine proteases) expressed in the salivary glands of the European tick Ixodes ricinus. Iripin-1, Iripin-3, and Iripin-8 were produced in the Escherichia coli expression system, and their effects on the immune response were evaluated using various in vitro assays and a mouse model of acute inflammation. Furthermore, the thesis involves one review article that summarizes the role of serpins in tick physiology and tick-host interaction.
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Functional analysis of tick salivary serine and cysteine protease inhibitors
KOTÁL, Jan
The proposed thesis focuses on the characterization of two protease inhibitors present in tick saliva. More specifically, the thesis presents immunomodulatory properties, biochemical specificity and structure of a cysteine protease inhibitor named Iristatin. Another characterized protein, IRS-8, comes from a serpin family (serine protease inhibitors) and inhibits blood coagulation and complement system in the host. Furthermore, the thesis provides a literature overview and discussion of tick salivary molecules in the context of tick-host-pathogen interaction, vaccination potential and medicine potential. Two review manuscripts, which are part of this thesis summarize the effects of tick saliva and protease inhibitors on host immune mechanisms.
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Inhibitors of proteolytic enzymes of Trematodes
Šteiger, Vladimír ; Kašný, Martin (advisor) ; Salát, Jiří (referee)
i Abstract Trematodes are important parasites possessing various localization in the bodies of invertebrate and vertebrate hosts, including human; therefore they are subject of long time intensive worldwide research. Trematodes developed various adaptations and strategies (some of them have also molecular background) enable them to survive in the host bodies. Trematodes produce large amount of different molecules, which are involved in various physiological processes. Inhibitors of proteolytic enzymes form a large group of biologically active compounds, e.g. they regulate the activity of peptidases or modulate host immune response. Many of these inhibitors are investigated as potential candidates in chemotherapeutic fight against trematodes. This thesis reviews the information concerning the natural inhibitors produced by trematodes and also synthetic inhibitors. Key words: Inhibitor, trematode, peptidase, serpin, cystatin i
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Inhibitors of proteolytic enzymes of Trematodes
Šteiger, Vladimír ; Kašný, Martin (advisor) ; Salát, Jiří (referee)
i Abstract Trematodes are important parasites possessing various localization in the bodies of invertebrate and vertebrate hosts, including human; therefore they are subject of long time intensive worldwide research. Trematodes developed various adaptations and strategies (some of them have also molecular background) enable them to survive in the host bodies. Trematodes produce large amount of different molecules, which are involved in various physiological processes. Inhibitors of proteolytic enzymes form a large group of biologically active compounds, e.g. they regulate the activity of peptidases or modulate host immune response. Many of these inhibitors are investigated as potential candidates in chemotherapeutic fight against trematodes. This thesis reviews the information concerning the natural inhibitors produced by trematodes and also synthetic inhibitors. Key words: Inhibitor, trematode, peptidase, serpin, cystatin i
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Preparation of recombinant inhibitor of serine proteases from the tick \kur{Ixodes ricinus}
VLNOVÁ, Ivana
Tick serine protease inhibitors could be important anti-tick vaccines targets because of their properties and functions. The aim of this work was to prepare recombinant inhibitor of serine proteases from the tick Ixodes ricinus in baculovirus expression system. Two tick saliva proteins of the serpine superfamily were selected for this purpose and transformed into plasmids. One recombinant protein was expressed in baculovirus expression system, purified and its biochemical analyses were done.
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The effect of tick´s serpin IRS-2 on dendritic cells activated by TLR4 ligand
POSPÍŠILOVÁ, Šárka
IRS-2 is the inhibitor of serine proteases from the Ixodes ricinus tick. My task in this thesis was to find out the effect of the IRS-2 on dendritic cells activated by TLR4 ligand or by Borrelia afzelii. This effect was studied on several levels. I focused on the cytokine production, the expression of costimulatory molecules and cell signaling pathways. The results show that the IRS-2 may inhibit the expression of costimulatory molecules CD-80 a CD-86 on the cell surface, but this finding needs to be confirmed again. The production of cytokines was not affected by the IRS-2. The effect of the IRS-2 on the activity of p38, Erk1/2 nor NF-?B in LPS stimulated cells vas not observed. The fosforylation of STAT 3 in cells activated by the B. afzelii was lowered by the IRS-2.
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Transcriptomic and functional analysis of salivary proteins from the tick \kur{Ixodes ricinus}
CHMELAŘ, Jindřich
This thesis was focused on the identification and characterization of the salivary proteins from Ixodes ricinus, the European vector of Lyme disease and tick-borne encephalitis causative agents. In the first part of this work, the the transcriptomic approach was used in order to identify and describe I. ricinus salivary proteins. The second part is dealing with functional and structural characterization of the salivary protein named IRS-2 (I. ricinus serpin-2).
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