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Salivary glycoproteins of bloodsucking arthropods
Sumová, Petra ; Volf, Petr (advisor) ; Mikeš, Libor (referee)
During obtaining their blood meal, bloodsucking arthropods salivate into their host. Bloodsucking arthropods' saliva contains wide array of bioactive macromolecules. Host organism develops antibody response against many of these molecules. Due to interspecies variability in salivary protein composition, detection of antibody response may serve as a marker of the exposure to individual species of bloodsucking arthropods. Host antibody response is mostly elicited by proteins or glycoproteins. Glycoproteins contain one or more oligosaccharide chains attached to the protein. Glycoprotein's antigenicity could be caused by either both parts, or by only the protein, or the sugar part. This fact has to be taken into consideration for choice of the expression system for recombinant glycoprotein synthesis. This work summarizes current knowledge about structure, function and features of salivary glycoproteins in various species of bloodsucking arthropods.
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Characterization and antigenic properties of salivary yellow-related proteins in phlebotomine sand flies
Cikrtová, Petra
2 ABSTRACT Yellow-related proteins (YRPs) form an abundant protein family, whose members were found in salivary gland transcriptomes of all sand fly species studied up to date. This protein family belongs to the group of MRJP/Yellow proteins occurring in insects and some other organisms. Though sharing similar folding as a six-bladed β-propeller comprising central tunnel, MRJP/Yellow proteins adopted different functions. The structure and biogenic amine-binding property described for the sand fly salivary YRPs was based on a single study conducted on Lutzomyia longipalpis. In the present work, we have modelled the structures of 32 salivary YRPs belonging to 13 different sand fly species. We have shown the general structural similarity of these proteins along with both inter- and intra-specific differences in surface charge, tunnel parameters and in amino acids composition of the amine-binding motif. These modifications indicated divergence in function of individual YRPs, which was experimentally verified in the second project focused on identification of the amine-binding properties of YRPs in two important vectors of Leishmania; Phlebotomus perniciosus and P. orientalis. In each species, two YRPs differ in affinities for biogenic amines serotonin, histamine and catecholamines. However, in both sand fly...
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Characterization and antigenic properties of salivary yellow-related proteins in phlebotomine sand flies
Cikrtová, Petra
2 ABSTRACT Yellow-related proteins (YRPs) form an abundant protein family, whose members were found in salivary gland transcriptomes of all sand fly species studied up to date. This protein family belongs to the group of MRJP/Yellow proteins occurring in insects and some other organisms. Though sharing similar folding as a six-bladed β-propeller comprising central tunnel, MRJP/Yellow proteins adopted different functions. The structure and biogenic amine-binding property described for the sand fly salivary YRPs was based on a single study conducted on Lutzomyia longipalpis. In the present work, we have modelled the structures of 32 salivary YRPs belonging to 13 different sand fly species. We have shown the general structural similarity of these proteins along with both inter- and intra-specific differences in surface charge, tunnel parameters and in amino acids composition of the amine-binding motif. These modifications indicated divergence in function of individual YRPs, which was experimentally verified in the second project focused on identification of the amine-binding properties of YRPs in two important vectors of Leishmania; Phlebotomus perniciosus and P. orientalis. In each species, two YRPs differ in affinities for biogenic amines serotonin, histamine and catecholamines. However, in both sand fly...
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Correlation of Immunohistochemical and Molecular Methods in Diagnostics of Salivary Gland Tumors
Horáková, Markéta ; Skálová, Alena (advisor) ; Laco, Jan (referee) ; Dušková, Jaroslava (referee)
This doctoral thesis is dealing with the correlation of morphological, immunohistochemical and genetical findings in malignant tumors of salivary glands. The first half of the thesis comprises the summary of current knowledge about salivary malignancies. The second half is presenting the research itself. The research results are divided into three parts. The first part is presenting the method of "2-step diagnostic test" of malignant tumors. This screening test aims to find new, so far not described gene aberrations with a focus on malignant tumors of salivary glands. This method takes place in two consecutive steps. In the first step the material is examined by an immunohistochemical mixture of antibodies, which non-specifically detects aberration in the genes NTRK1-3, ALK and ROS1. In the second step all positive cases are subjected to highly sensitive and specific molecular-genetic examination by the method of next generation sequencing (NGS) using the Archer kit. In the second part of the work there has been designed the approach to the cytological diagnosis of salivary secretory carcinoma by the fine-needle aspiration (FNA). This part is describing to the details the cytomorphology of secretory carcinoma in both, Pap smears and cell blocks, from which additional immunocytochemical and genetic...
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Diferenciální exprese transkriptů slinných žláz klíštěte \kur{Ixodes ricinus}
KROPÁČKOVÁ, Sára
Ticks are using different mechanisms for host immune system suppression. In this thesis, I compared ten transcripts from ticks salivary glands, which are differentially expressed during feeding on live animal and artificial systems. Further, a tick protein anticomplement 1, AC1, was chosen and subjected to deeper investigation, because of the significant difference in expression during feeding. Serum with specific antibodies was made and AC1 expression levels were examined in nymphs and adults, during feeding on different hosts. Moreover, any potential interacting partners from host serum through covalent binding were tested. This study contributes to the understanding of regulation of expression and funtional characterisation of tick salivary proteins.
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Salivary glycoproteins of bloodsucking arthropods
Sumová, Petra ; Volf, Petr (advisor) ; Mikeš, Libor (referee)
During obtaining their blood meal, bloodsucking arthropods salivate into their host. Bloodsucking arthropods' saliva contains wide array of bioactive macromolecules. Host organism develops antibody response against many of these molecules. Due to interspecies variability in salivary protein composition, detection of antibody response may serve as a marker of the exposure to individual species of bloodsucking arthropods. Host antibody response is mostly elicited by proteins or glycoproteins. Glycoproteins contain one or more oligosaccharide chains attached to the protein. Glycoprotein's antigenicity could be caused by either both parts, or by only the protein, or the sugar part. This fact has to be taken into consideration for choice of the expression system for recombinant glycoprotein synthesis. This work summarizes current knowledge about structure, function and features of salivary glycoproteins in various species of bloodsucking arthropods.
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Salivary gland hyaluronidase of tabanids and ticks
Tothová, Viktorie ; Volf, Petr (advisor) ; Mikeš, Libor (referee)
6 1. Abstrakt Hyaluronidázy jsou významnou skupinou enzymů odpovědných za štěpení kyseliny hyaluronové, která je jednou z hlavních složek pojivové tkáně obratlovců. U většiny krevsajících členovců je tento enzym přítomen ve slinách a usnadňuje sání tím, že se podílí na průniku kůží a zvětšení potravní léze v místě bodnutí. Vzniklé fragmenty extracelulární matrix mohou navíc modulovat lokální imunitní odpověď hostitele a zvyšovat pravděpodobnost přenosu patogenů. V naší práci jsme se zaměřili na průkaz hyaluronidázové aktivity u klíštěte Ixodes ricinus (Acari: Ixodidae) a zástupců čtyř rodů ovádů (Diptera: Tabanidae). V slinných žlázách klíšťat se hyaluronidázovou aktivitu detekovat nepodařilo. Naopak, ve slinných žlázách ovádů jsme prokázali velmi silnou aktivitu u všech studovaných druhů a dále ji charakterizovali biochemickými a elektroforetickými metodami. Hyaluronidázy ovádů štěpí hyaluronan i chondroitin sulfát; enzymy jednotlivých druhů se mírně liší molekulovou hmotností, pH optimem a citlivostí k redukujícím podmínkám. Abstract (in English) Hyaluronidases are an important group of enzymes responsible for cleaving hyaluronic acid, which is a major component of the extracellular matrix of vertebrates. In bloodsucking arthropods these enzymes are frequently present in saliva. Salivary hyaluronidases...
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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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