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Benkovská, Dagmar. "Proteomické studie ječmene související s výrobou piva." Doctoral thesis, Vysoké učení technické v Brně. Fakulta chemická, 2013. http://www.nusl.cz/ntk/nusl-233378.
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Tato práce se zabývá proteomickými studiemi ječmene v souvislosti s výrobou piva. Ječmen patří mezi nejvýznamnější plodiny na světě a je využíván hlavně pro sladovnické účely, nejčastěji pro pivovarnictví. Studium proteinů ječmene během sladování a výroby piva poskytuje informace o změnách v proteinovém složení nebo jejich posttranslačních modifikacích. Jelikož jsou proteiny v ječmeni a jejich změny zásadní pro kvalitu sladu a piva, proteomické studie ječmene mají potenciál pro zlepšení procesu sladování a pivovarnictví. Hlavním cílem této práce je studium ve vodě rozpustných proteinů ječmene a jejich změn, ke kterým dochází během sladování a výroby piva. Rozdíly v proteinovém složení byly sledovány pomocí gelové elektroforézy, kapalinové chromatografie na reverzní fázi, gelové chromatografie a MALDI-TOF hmotnostní spektrometrie. Během sladování se vlivem klíčení zrna zvyšuje množství některých proteinů a také jsou tvořeny nové proteiny. V průběhu vaření piva se naopak v důsledku vysoké teploty a enzymatické aktivity proteáz mnoho proteinů rozkládá. Těmto drsným podmínkám odolají jen některé proteiny, které přechází až do piva a mohou ovlivnit jeho kvalitu. Dále byly zkoumány různé odrůdy ječmene a jejich rozdíly. Byly porovnány odrůdy povolené pro výrobu certifikovaného Českého piva s jednou osvědčenou sladovnickou odrůdou a jednou nesladovnickou odrůdou ječmene. Kromě toho byly studovány v alkoholu rozpustné proteiny ječmene a jejich změny v průběhu sladování. Zvláštní pozornost byla věnována vybrané skupině posttranslačních modifikací proteinů: glykosylacím. Neenzymaticky glykosylované proteiny ječmene (neboli glykované proteiny) jsou tvořeny v průběhu sladování kvůli přítomnosti velkého množství glukózy uvolněné z rozkladu škrobu. Glykované proteiny ovlivňují stabilitu proteinů a kvalitu piva, obzvlášť pěnotvorný účinek. Enzymatické N-glykosylace představují nejčastěji studované posttranslační modifikace u rostlin, protože glykoproteiny hrají klíčovou roli v různých biologických funkcích. Glykoproteiny jsou často přítomny v malém množství, a proto je pro jejich analýzu potřebné obohacení glykoproteinů z komplexní směsi. Pro studium glykoproteinů byla využita afinitní chromatografie s lektinem concanavalin A. Kromě toho byla také optimalizována analýza sacharidové části glykoproteinů. Tato disertační práce přináší důležité informace o proteinech ječmene, jejich změnách a analýze, které budou užitečné pro další studium.
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Seifermann, Marco [Verfasser]. "Die Regulation der Transkription proinflammatorischer Zytokine durch oxidative Basenmodifikationen und die DNA-Reparatur Glykosylase OGG1 / Marco Seifermann." Mainz : Universitätsbibliothek Mainz, 2017. http://d-nb.info/1138951064/34.
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Köger, Nicole [Verfasser]. "Analyse der Promotorregulation und des Effekts humaner genetischer Varianten auf die Expression der DNA-Glykosylase MUTYH / Nicole Köger." Gießen : Universitätsbibliothek, 2018. http://d-nb.info/1162840064/34.
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Sommer, Tina Maria [Verfasser]. "Einfluss der DNA-Glykosylase OGG1 auf die TGF beta-vermittelte Induktion einer Epithelial-mesenchymalen Transition (EMT) / Tina Maria Sommer." Mainz : Universitätsbibliothek Mainz, 2018. http://d-nb.info/1162143193/34.
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Grombacher, Thomas [Verfasser]. "Reparatur von Methylierungsschäden der DNA: Konstitutive Expression und Induzierbarkeit der O⁶-Methylguanin-DNA-Methyltransferase und der N-Methylpurin-DNA-Glykosylase in Säugerzellen / Thomas Grombacher." Karlsruhe : KIT-Bibliothek, 1995. http://d-nb.info/1124902686/34.
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Vogel, Julia [Verfasser], and Eric [Akademischer Betreuer] Metzen. "Die Rolle des Basen-Exzisionsreparaturenzyms 8-Oxoguanin DNA Glykosylase 1 (OGG1) in Kombination mit Mut-T Homolog-1 (MTH1) Inhibition / Julia Vogel ; Betreuer: Eric Metzen." Duisburg, 2021. http://d-nb.info/1236501896/34.
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Mehringer, Christian Felix [Verfasser], Ingrid [Gutachter] Tessmer, and Antje [Gutachter] Gohla. "Optimierung und Objektivierung der DNA-Biegewinkelmessung zur Untersuchung der initialen Schadenserkennung von Glykosylasen im Rahmen der Basen-Exzisions-Reparatur / Christian Felix Mehringer ; Gutachter: Ingrid Tessmer, Antje Gohla." Würzburg : Universität Würzburg, 2021. http://d-nb.info/1230324194/34.
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Albahri, Ziad. "Záchyt vrozených poruch glykosylace." Doctoral thesis, 2006. http://www.nusl.cz/ntk/nusl-266055.
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Ondrušková, Nina. "Studium transferinu jako markeru dědičných poruch glykosylace." Master's thesis, 2010. http://www.nusl.cz/ntk/nusl-296231.
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Congenital disorders of glycosylation (CDG) represent a heterogeneous group of mul- tisystemic metabolic disorders which are caused by defects in biosynthetic pathways of glycoproteins. The screening test for N-glycosylation disorders is the analyses of sialylated isoforms of serum transferrin (Tf) by means of isoelectric focusing (IEF). Two distinct pathological IEF patterns of Tf are observed. A type I pattern is cha- racterized by a decrease of tetra- and an increase of di- and asialotransferrin, whereas a type II pattern shows in addition an increase of tri- and monosialotransferrin. The aims of diploma thesis were: 1) to evaluate reference range for spectrum of sialylated forms of Tf separated by IEF and 2) to perform biochemical and molecular analyses in three patients (P1-P3) with clinical suspicion for CDG. Serum and genomic DNA from three patients with clinical suspicion for CDG and family members of P1 were analysed. Sera from 99 healthy volunteers within the age range of 2-42 years served as a control group. Tf was analysed by IEF with direct immunofixation, SDS-PAGE and Western blot using specific antibody against human Tf (Dako). Profiles of Tf were quantified by AlphaEaseFC software (Alpha Innotech). Data were analysed by software STATISTICA 9.0 (StatSoft). TF a PMM2 genes were analysed...
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Hůzová, Zuzana. "Glykosylace, ultrastruktura a antigeny povrchu larev motolic." Master's thesis, 2006. http://www.nusl.cz/ntk/nusl-370209.
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Ondrušková, Nina. "Studium dědičných poruch glykosylace na biochemické a molekulární úrovni." Doctoral thesis, 2016. http://www.nusl.cz/ntk/nusl-353578.
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Congenital disorders of glycosylation (CDG) represent a rapidly growing group of rare inherited metabolic diseases with estimated prevalence as high as 1:20 000, which are caused by genetic defects that impair the process of glycosylation, i.e. the enzymatic addition of a specific saccharide structure onto a protein or lipid backbone. Due to non-specificity and variability of clinical symptoms in the patients, the medical diagnosis of CDG remains extremely challenging and significantly relies on accurate biochemical and genetic analyses. The overall goal of the present dissertation thesis was to study CDG at the biochemical and molecular genetic level in the context of the Czech and Slovak Republic, which involved three specific aims: A.) to introduce and optimize laboratory screening methods for CDG detection in a group of clinically suspected patients, B.) to determine the corresponding genetic defect in the positive patients selected via CDG screening and to study the pathobiochemical aspects of specific CDG types at the cellular level, and C.) to analyze glycosylation disturbances of non- CDG etiology. Contributions of this work include optimization of isoelectric focusing of apolipoprotein C-III (ApoC-III) as a screening method for O-glycosylation abnormalities, as well as the description of...
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Lejsalová, Alžběta. "Vliv O-glykosylace v onkofetálním fibronektinu na kostní formaci." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-297637.
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Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Performed at Institute of Immunology The Ruprecht-Karls-University Heidelberg Candidate: Alžběta Lejsalová Supervisor: Doc. PharmDr. Tomáš Šimůnek, Ph.D. Specialized supervisor: Prof. Dr. Inaam Nakchbandi Title of diploma thesis: The Influence of the O-glycosylation in oncofetal fibronectin on the bone formation Hepatic osteodystrophy is a major complication in patients with chronic cholestatic liver diseases which significantly contributes to decreased quality of life. It was established that increased levels of the oncofetal domain of fibronectin correlated with bone loss and decrease in bone formation in patients with primary biliary cirrhosis. The oncofetal fibronectin is characterized by the O-glycosylation in the variable domain. Another O-glycosylation is found on the NH2-terminus. The aim of this project was to prepared O-deglycosylated fibronectin isoforms and fibronectin knockdown mammalian cells for further research on the effect of O-glycosylation on the bone formation.
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Veselá, Šárka. "Využití průtokové cytometrie pro diagnostiku a charakterizaci dědičných poruch glykosylace." Master's thesis, 2021. http://www.nusl.cz/ntk/nusl-448756.
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Congenital disorders of glycosylation (CDG) are rare multisystem metabolic diseases and their number has rapidly grown in recent years. The clinical manifestation includes very broad spectrum of symptoms. In most of all cases CDG are caused by mutations in genes encoding the enzymes of glycosylation pathway. Based on the type of defect, CDG are divided into the following groups: disorders of N-glycosylation or O-glycosylation of proteins, defects in modification of proteins by GPI anchor, disorders of lipid glycosylation and defects that impact multiple glycosylation pathways. The aim of the thesis was to find new biochemical analyses suitable for diagnostics and characterization of CDG patients. The experimental conditions were optimized for selected markers (Sambucus Nigra (SNA) lectin, proaerolysin (FLAER), antibodies to proteins CD55 and CD59) and the staining was applied to cultivated skin fibroblasts from controls and patients diagnosed with CDG by whole-exome sequencing (ATP6AP1-CDG, PIGN-CDG, SLC10A7-CDG, PISD deficiency). The experiments were performed using flow cytometry (FACS) and fluorescent microscopy (FM). The detection of sialylation by SNA lectin and analysis of the mitochondrial membrane potential changes by a fluorescent labelled probe JC-1 with FCCP simulation of mitochondrial...
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Sumová, Petra. "Glykosylace a antigenní vlastnosti proteinů ze slin flebotomů Phlebotomus perniciosus a P. orientalis." Master's thesis, 2014. http://www.nusl.cz/ntk/nusl-340884.
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The goal of this study was to map the glycosylation pattern and antigenic properties of the salivary proteins of two closely related sand fly species, Phlebotomus perniciosus and P. orientalis. Affinity blotting with commercially available lectins revealed that many salivary proteins of these species are N-glycosylated, while the presence of O-glycosylation could not be confirmed. The level of N-glycosylation of most of these proteins is quite low, a larger number of potential N-glycosylation sites were found only in the amino acid sequences of P. orientalis hyaluronidase and endonucleases of both species tested. Four antigens from P. perniciosus salivary glands were selected for expression in a bacterial expression system; two of these proteins (PpeSP01 and PpeSP01B) were not glycosylated and the glycosylation level of the remaining two (PpeSP03B and PpeSP07) was low. The antigenic properties of the four chosen recombinant proteins were subsequently tested using immunoblot and ELISA. During the initial experiments with the sera of dogs experimentally bitten by P. perniciosus, two proteins (rSP07 and rSP01B) were proven unsuitable and they were excluded from further experiments. Recombinant proteins rSP03B and rSP01 were recognized by the same IgG antibodies as the native forms of these proteins...
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Zdražilová, Lucie. "Analýza obsahu dolicholu v moči u pacientů s dědičnými poruchami glykosylace pomocí hmotnostní spektrometrie." Master's thesis, 2018. http://www.nusl.cz/ntk/nusl-379509.
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Dolichol is a membrane lipid, which carries monnosaccharides and glycans for N-linked protein glycosylation and glycosylphosphatidylinositol-anchor biosynthesis occuring in endoplasmic reticulum. Its structure is composed of isoprenoid units. Dolichol is present in all tissues and in most of the membrane organelles of eukaryotic cells. Recently some types of congenital disorders of glycosylation have been described as a consequence of dolichol biosynthesis and metabolism defects, which are not detectable by standard methods. The aim of this diploma thesis was to analyze dolichol content in urine and in different tissues from patients with deficiency in dolichol biosynthesis by mass spectrometry and to study the impact of these defects on energetic metabolism. Biological material for this study consisted of urine samples from 76 controls with age ranging from 1 months to 81 years, 6 patients with congenital disorders of glycosylation and 43 patients with suspicion of congenital disorder of glycosylation; samples of frontal cortex, liver, muscle and heart tissues from 2 patients with mutation in NUS1 gene and controls. Urine samples were stored at -20 řC and tissue homogenates were stored in -80 řC until analysis. Lipid fraction after extraction was separated by liquid chromatography. Dolichols were...
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Sedláková, Jana. "Syntéza fosforamidátových proléčiv "ProTides" jako nových potenciálních léčiv pro terapii vrozených poruch glykosylace a syndromu vyčerpání mitochondriální DNA." Master's thesis, 2018. http://www.nusl.cz/ntk/nusl-380907.
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Charles University Faculty of Pharmacy in Hradec Králové Department of Organic And Bioorganic Chemistry Candidate: Jana Sedláková Supervisors: Dr. Fabrizio Pertusati doc. PharmDr. Jaroslav Roh, Ph.D. Title of diploma thesis: Synthesis of phosphoramidate prodrugs "ProTides" as novel potential therapeutic agents for the treatment of congenital disorders of glycosylation and mitochondrial DNA depletion syndrome At the present time, no effective treatment is available neither for the most of the congenital disorders of glycosylation (CDGs) nor the mitochondrial DNA depletion syndrome (MDS). Regarding the CDG therapy, D-mannose-1-phosphate (Man-1-P) offers considerable pharmacological potential to improve the pathological patterns in patients affected by phosphomannomutase 2 deficiency (PMM2-CDG), similarly as N-acetyl-D-mannosamine-6-phosphate (ManNAc-6-P) in case of GNE myopathy (GNEM). Administration of selected deoxyribonucleotides was proposed as a potential pharmacological strategy for the treatment of MDS. Unfortunately, the problematic membrane penetration of such polar molecules reduces their effect and limits their clinical application. Hydrophobic, membrane permeable derivatives of the sugar monophosphates and nucleotides, might represent more efficient potential therapeutics for CDGs and...
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FIŠER, Miroslav. "Identifikace a biochemická charakterizace lektinů v hemolymfě tří druhů klíšťat rodu \kur{Rhipicephalus}." Master's thesis, 2009. http://www.nusl.cz/ntk/nusl-49953.
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Lectins are tissue specific carbohydrate binding proteins with possible functions in invertebrate immunity and pathogen transmission. The main goal of this study was to identify hemolymph lectins in three different tick species. Three proteins with molecular weights of 58 kDa, 75 kDa and 180 kDa were detected in all investigated species using antibodies directed against hemagglutination activity of Rhipicephalus appendiculatus hemolymph. These proteins were characterized by biochemical methods such as Schiff/periodate staining, lectin blotting, enzymatic deglycosylation, hemagglutination analysis, immunoblotting, and mass spectrometry.
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Pažický, Samuel. "Studium oligomerizace proteinu NKp30 a jeho interakce s B7-H6." Master's thesis, 2016. http://www.nusl.cz/ntk/nusl-343094.
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NK cells are important part of immune system, recognizing and eliminating tumor cells and cells infected by viruses. For the target cell recognition, binding of ligands by activating receptors plays a crucial role. Activating receptor NKp30, protein of family of natural cytotoxicity receptors, is able to bind multiple ligands either present on tumor cell surface or being part of some viruses. B7-H6 is one of the ligands of NKp30 and its specific constitutive expression on some tumor cells and cell lines makes it an interesting biological target. Although the NKp30/B7-H6 complex structure has been solved, structural basis of some important features of their binding is not explained yet. Soluble form of NKp30 receptor binding domain creates oligomers, presence of which is dependent on C-terminus length of its domain and its N-glycosylation; however, structural insight into formation of the oligomers and their significance is not known. Furthermore, binding affinity of NKp30 to its ligands is dependent on presence of its glycosylation and glycosylation type. We have already found out that NKp30 oligomerization is dependent on its glycosylation. In my work, I attempted to gain detailed functional and structural information about oligomerization of NKp30 and its binding to B7-H6 by multimethodical...
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Downey, Alan Michael. "Vývoj nových glykosylačních metod pro syntézu nukleosidů." Doctoral thesis, 2017. http://www.nusl.cz/ntk/nusl-371014.
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As they make up DNA and RNA, nucleosides are considered the key to life. Synthetic nucleosides also constitute many drugs that treat viral infections and cancer. As a result, more efficient methods to access these crucial molecules would have implications that extend beyond a synthetic chemist's benchtop and into medicinal chemistry and medical research. One of the most challenging steps in the synthesis of nucleosides is the glycosylation step between the acceptor heterocycle (nucleobase) and the saccharide-based donor. Often to obtain satisfactory yield of this step with good regio- and stereochemical control the extensive use of protecting groups must be employed to squelch reactivity at unwanted reactive groups. Consequently, this process of protection−glycosylation−deprotection is laborious, inefficient, and often requires the use of toxic reagents. It would be, therefore, highly welcomed if new methodology to effect this glycosylation step was designed that reduces or removes the need to use protecting groups, but would still provide nucleosides in good yield, regio- and stereoselectively. Herein, this thesis presents my efforts into achieving this end. By employing modified Mitsunobu conditions, I determined that it is possible to directly glycosylate a nucleobase with D-ribose to afford...
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Skřenková, Kristýna. "Molekulární mechanismy regulace transportu a funkce různých podtypů NMDA receptorů v hipokampálních neuronech." Doctoral thesis, 2020. http://www.nusl.cz/ntk/nusl-411734.
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of Ph.D. thesis Molecular mechanisms of regulation of trafficking and function of different subtypes of NMDA receptors in hippocampal neurons Mgr. Kristýna Skřenková N-methyl-D-aspartate (NMDA) receptors are ionotropic glutamate receptors that play a key role in the mammalian central nervous system. Under physiological conditions, these receptors are important for excitatory synaptic transmission and memory formation. However, under pathological conditions, their abnormal regulation or activation may lead to many neurological and psychiatric disorders, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, epilepsy or schizophrenia. Previous studies have shown that the number and type of NMDA receptors on the cell surface are regulated at multiple levels, including their synthesis, folding, internalization or degradation. During the trafficking of NMDA receptors to the cell surface membrane, both the agonist binding and receptor activation are examined. Moreover, NMDA receptors undergo many posttranslational modifications such as palmitoylation, phosphorylation or N-glycosylation. In this thesis, we studied the molecular mechanisms that may affect the trafficking and functional properties of NMDA receptors in mammalian cells and rat hippocampal neurons. Specifically, we studied i)...
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Libigerová, Martina. "Studium molekulárních mechanismů eliminace klinicky významných nádorů zabíječskými buňkami." Master's thesis, 2010. http://www.nusl.cz/ntk/nusl-285215.
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Carbohydrates have an essentials role in wide range of biological phenomena. It is well known that most of the eukaryotic proteins are glycosylated and that their glycosylation undergoes dynamic changes, nevertheless the biological imperative for these modifications is still not fully understood. However, one area in which the importace of cell surface glycosylation has recently been the subject of active investigations is the tumor plasma membrane biology, where many changes in glycosylation have been found useful for diagnosis, and mostly recent, even for the therapies of malignant disease. Interestingly cell surface glycoconjugates, namely N-linked and O-linked oligosaccharides have been found therapeutically attractive for treatment of certain tumors. And although our understanding of the participation of these principal glycan classes in tumorigenesis is far from complete, there are already several examples of carbohydrate-based antitumor vaccines. Therefore, we decided to give this issue more attention, especially the molecular mechanisms responsible for identifying changes in glycosylation of the surface of tumor cells of the immune system. Although in the past in our laboratory identified a receptor-type lectin specific lectin receptors on natural killer cells, very little is yet known...
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Vomelová, Ivana. "Charakterizace transgenních forem dipeptidylpeptidasy IV exprimovaných v astrocytární buněčné linii U373MG." Master's thesis, 2010. http://www.nusl.cz/ntk/nusl-285106.
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Dipeptidyl peptidase IV (DPP-IV) is a serine protease, which executes its proteolytic activity by cleaving X-Pro dipeptides from the N-termini of its substrates. Furthermore, DPP-IV exhibits many biological functions independent of its enzymatic aktivity. Previous studies in our laboratory proved increased expression of DPP-IV in high-grade astrocytic tumours. To evaluate the enzymatic and non-enzymatic functions of DPP-IV in a glioma model, clones of asctrocytic cell line U373MG transfected by enzymatically inactive, mutated DPP-IV (mutDPP-IV) and enzymatically active, wild type DPP-IV (wtDPP-IV), were prepared. Enzymatically inactive mutDPP-IV was prepared using point mutation the active site serine residue. Cells U373MG were transfected using a doxycycline inducible Tet-On® system. For further analysis of the transgenic forms of DPP-IV, methods were used for verification of protein expression, enzymatic activity and subcellular localization. Doxycycline induced U373MG mutDPP-IV and U373MG wtDPP-IV cells, expressing mutated and wild type DPP-IV, respectivelly, exhibited increased expression of transgenic DPP-IV in a concentration and time dependent manner. Doxycycline induced U373MG wtDPP-IV cells exhibited both increased expression and enzymatic activity of DPP-IV. In contrast, DPP-IV enzymatic...
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ŠTĚRBOVÁ, Jarmila. "Hemlipoglykoprotein z hemolymfy klíštěte \kur{Dermacentor marginatus}: purifikace a biochemická charakterizace." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-54463.
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The aim of the study is the purification and characterization of hemelipoglycoprotein, a carrier protein from the ornate sheep tick Dermacentor marginatus. The protein was characterized by biochemical methods with emphasis on its glycosylation and native molecular weight, and examining its carbohydrate-binding specificity.
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de, Sousa Santos Abreu Celeste. "Interakce Galektinu-1 s receptory lidských NK buněk." Master's thesis, 2019. http://www.nusl.cz/ntk/nusl-403989.
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Natural killer (NK) cells are a subpopulation of effector lymphocytes with cytotoxic activity and cytokine-producing functions considered as an integral part of the innate immune response. Functions of NK cells include tumour elimination, engagement and regulation of antiviral immune responses and regulation of immune cells by production and secretion of chemokines and cytokines. CD69 is a C-type lectin-like transmembrane receptor expressed in NK cells. CD69 is an activating receptor and acts also as a very early marker of lymphocyte activation. Putative protein ligands have been described for CD69 in the last years: Galectin-1, S1P1, S100A8/S100A9 and Myl9/12. Galectin-1 is a prototypical lectin characterized by the presence of a common lectin structural fold and a carbohydrate recognition domain involved in carbohydrate binding. Galectin-1 was identified as a binding partner for CD69 based on biological and functional studies, but structural details about the complex are still missing. This thesis describes the successful establishment of an expression protocol for a tag-less cysteine-less mutant of galectin-1 and the study of the interaction between galectin-1 and NK cell receptors. The interaction was studied using microscale thermophoresis and confirmed as dependent on the presence of a...
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STROUHALOVÁ, Renata. "Změny v genomu viru klíšťové encefalitidy u variant s různou historií pasáží a odlišnými biologickými vlastnostmi." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-53938.
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Tick-borne encephalitis virus (strain Hypr) was serially subcultured in PS cells and tick cell line IRE/CTVM19, producing four different viral variants. Biological properties of these new variants were investigated in mouse model. Possible determinants of virulence were found by full-genome sequencing. The role of glycosylation for tick-borne encephalitis virus was evaluated.
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ŠIMONOVÁ, Zuzana. "Určení N-glykomu klíštěte \kur{Ixodes ricinus} a \kur{Dermacentor marginatus}; analýza N-glykanů v tkáních klíštěte a jejich porovnání." Master's thesis, 2011. http://www.nusl.cz/ntk/nusl-85075.
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Glycosylation in vertebrates has a main role in many important processes such as cell transport, protein folding, secretion of proteins etc. What function has glycosylation in arthropods, for example in ticks, is rarely studied. This work was focused on analysis of N-glycans in tick tissues, namely in Ixodes ricinus and Dermacentor marginatus. High-mannose glycans as well as complex glycans with or without core-fucosylation were identified in this study.Furthermore several sialylated glycans were present in the studied samples. Sialic acid is found in arthropods rarely and this is the first study which directly proves its presence in ticks using mass spectrometry.
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Hemelíková, Katarína. "Regulace transportu NMDA receptorů v savčích neuronech." Doctoral thesis, 2018. http://www.nusl.cz/ntk/nusl-389420.
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N-methyl-D-aspartate (NMDA) receptors are a subclass of glutamate receptors that play an essential role in mediating excitatory neurotransmission and synaptic plasticity in the mammalian central nervous system (CNS). The activation of NMDA receptors plays a key role in brain development and memory formation. Abnormal regulation of NMDA receptors plays a critical role in the etiology of many neuropsychiatric disorders. NMDA receptors form a heterotetrameric complex composed of GluN1, GluN2(A-D) and GluN3(A, B) subunits. The NMDA receptors surface expression is regulated at multiple levels including early processing (synthesis, subunit assembly, endoplasmic reticulum (ER) processing, intracellular trafficking to the cell surface), internalization, recycling and degradation. NMDA receptors are regulated by the availability of GluN subunits within the ER, the presence of ER retention and export signals, and posttranslational modifications including phosphorylation and palmitoylation. However, the role of N-glycosylation in regulating of NMDA receptor processing has not been studied in detail. The aim of this study was to clarify the mechanisms of regulation of surface expression and functional properties of NMDA receptors. We used a combination of molecular biology, microscopy, biochemistry and...
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Darebná, Petra. "Charakterizace glykoforem haptoglobinu v lidském séru." Master's thesis, 2015. http://www.nusl.cz/ntk/nusl-353849.
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Characterization of haptoglobin glycoforms in human serum Petra Darebná (Katedra biochemie, Přírodovědecká fakulta, Univerzita Karlova v Praze, Česká republika) Changes in glycosylation of proteins are associated with several types of cancer, including hepatocelular carcinoma and colorectal carcinoma. This project deals with data independent analysis using ion cyclotron resonance with Fourier transform and tandem mass spectrometry with liquid chromatogramy and multiple reaction monitoring to quantify these changes in hepatocelular cancinoma and colorectal carcinoma with liver metastases. In the first part of the project the haptoglobin was enriched from pooled serum samples of pacients with hepatocellular carcinoma, colorectal cancer and colorectal carcinoma with metastases using hemoglobin immobilized on CNBr-activated Sepharose 4B and then purified by high pressure liquid chromatography. Individual haptoglobin glycopeptides were analyzed using ion cyclotron resonance with Fourier transform. In the second part of the project we analyzed changes in glycosylation depending on diferent tumor diseases in partially depleted serum of individual patients using ethanol precipitation. Individual fucosylated glycoforms of N-glycopeptides of serum proteins were compared with their nonfucosylated forms. In...
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Starkuviene, Vytaute. "Identifizierung und Charakterisierung von thermostabilen Uracil Glykosylasen von Archaeon Methanobacterium thermoautotrophicum und Bakterium Thermus thermophilus." Doctoral thesis, 2001. http://hdl.handle.net/11858/00-1735-0000-0006-AC13-A.
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Mehringer, Christian Felix. "Optimierung und Objektivierung der DNA-Biegewinkelmessung zur Untersuchung der initialen Schadenserkennung von Glykosylasen im Rahmen der Basen-Exzisions-Reparatur." Doctoral thesis, 2021. https://doi.org/10.25972/OPUS-23084.
Full textAbstract:
Im Rahmen dieser Doktorarbeit sollte anknüpfend an die Ergebnisse aus vo-rangegangenen Untersuchungen der AG Tessmer, das von Büchner et al. [1] vorgestellte Modell zur DNA-Schadenserkennung, welches im Speziellen auf Daten zu den Glykosylasen hTDG und hOGG1 basierte, auf seine Allgemein-gültigkeit für DNA-Glykosylasen untersucht werden. Das Modell beschreibt den Prozess der Schadenserkennung als eine notwendige Übereinstimmung der passiven Biegung am Schadensort mit dem aktiven BiegungswinkeI der scha-densspezifischen Glykosylase. Ein wesentlicher Bestandteil dieser Arbeit war zudem die Etablierung einer automatisierten Messsoftware zur objektiven Biegewinkelmessung an DNA-Strängen in rasterkraftmikroskopischen Aufnah-men. Dies wurde mit verschiedenen Bildverarbeitungsprogrammen sowie einer in MATLAB implementierten Messsoftware erreicht und das Programm zudem auf die Biegewinkelmessung von proteininduzierten Biegewinkeln erweitert. Zur Anwendung kam die Methode der automatisierten Biegewinkelmessung sowohl an rasterkraftmikroskopischen Aufnahmen der Glykosylase MutY gebunden an ungeschädigter DNA als auch an Aufnahmen von DNA mit und ohne Basen-schaden. Neben oxoG:A und G:A, den spezifischen MutY-Zielschäden, wurden auch andere Basenschäden wie beispielsweise oxoG:C und ethenoA:T vermes-sen und zudem die von der Glykosylase MutY an ungeschädigter DNA induzier-te Biegung mit den Biegewinkeln der jeweiligen Zielschäden verglichen. Die Übereinstimmung in den Konformationen der Zielschäden und der Reparatur-komplexe auch für die Glykosylase MutY (wie bereits für hTDG und hOGG1 in oben genannter Arbeit gezeigt) erlauben ein verbessertes Verständnis der Schadenssuche und -erkennung durch DNA-Glykosylasen, indem sie die All-gemeingültigkeit einer Biegungsenergie-basierten initialen Schadenserkennung durch DNA-Glykosylasen unterstützen. Die etablierte Messsoftware kann zu-künftig an weiteren DNA-Schäden und den entsprechenden Protein-DNA-Komplexen ihre Anwendung finden und kann somit durch die effektive Gewin-nung objektiver Daten in großer Menge zur Stützung des Modells beitragenThe focus of this thesis was to test the general applicability of a model for initial lesion detection by base excision repair (BER) glycosylases. This thesis built on previous results from the Tessmer laboratory on the human base excision re-pair (BER) glycosylases hTDG and hOGG1 (Büchner et al. [1]). Based on this work, a model for initial lesion detection by glycosylases had been proposed that describes the process of damage recognition as a necessary match of the passive bending at the point of damage with the active bending by the damage-specific glycosylase. An essential component of this work was also the estab-lishment of an automated measurement software for objective bend angle measurements on DNA strands in atomic force microscopy (AFM) images. This was achieved with various image processing programs and a custom written MATLAB software. In addition, the procedure was extended to the measure-ment of DNA bend angles in protein-DNA complexes. In particular, the automa-ted bend angle analsyis was applied to AFM images of the glycosylase MutY bound to non-specific DNA and MutY target lesions (oxoG:A and G:A), as well as other DNA damages (oxoG:C and ethenoA:T). In the analyses, DNA bending induced by MutY in undamaged DNA was measured and compared to bending at the respective target damage. Similarities in the conformations of target da-mage and repair complexes also for this additional glycosylase (as already shown for hTDG and hOGG1 in above mentioned work) allow an improved un-derstanding of DNA glycosylase damage search and recognition by supporting the general validity of bending energy-based initial damage detection by DNA glycosylases. In addition, the established measurement software can also be used to measure DNA bending by other protein systems in an unbiased manner and on a high-throughput scale. The software thus contributes to the effective acquisition of objective data
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Landová, Barbora. "Strukturní studie mechanismů opravy poškozené DNA Nei glykosylasou." Master's thesis, 2019. http://www.nusl.cz/ntk/nusl-397027.
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Abasic sites (Ap site, from apurinic/apyrimidinic) are one of the most common lesions generated in DNA by spontaneous base loss or DNA repair processes. There are two equilibrating forms of an Ap site - ring-open aldehyde and cyclic hemiacetal. Ring- opened aldehydes are reactive electrophilic groups capable of formation covalent adduct with nucleophilic sites in DNA. DNA interstrand cross-link (ICL) resulting from the Ap sites is formed spontaneously as a covalent bond between ring-open aldehyde and amin group of adenin residue in the opposite strand of double stranded DNA. ICLs block DNA replication and transcription. The formation of Ap site derived ICL is relatively long process taking several hours. We assume that the ring-opening of an abasic site is the rate-limiting step in the formation of the thermodynamic ICL. However, formation, stability and DNA repair of Ap-ICL are still poorly understood processes. Here, I have set up mechanistic in vitro experiments to reveal and calculate the probability of Ap-ICl formation in vivo. In more detail, I study the rates of formation of Ap-ICLs in the sequence context of neighbouring nucleotides of freshly formed covalent bond of ICL. I focus on sequence preference, the influence of AT/ GC rich regions and the length of oligonucleotides. I have...