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Binding of low-molecular weight ligands to serum albumin
Buša, Michal ; Hudeček, Jiří (advisor) ; Vaněk, Ondřej (referee)
In this Thesis, we studied the possibilities of an enlargement of the existing practical task in the Advanced practical course in biochemistry I, dealing with interaction of a dye (bromophenol blue) with serum albumin. Experimental protocol was proposed and tested, allowing the inclusion of Scatchard plot (in addition to the previously used Job plot). As part of the Thesis, the new protocol for students is presented, which may help them to understand the most used approaches for studying the stoichiometry of protein-ligand complexes. In Czech.
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Protein-nanodiamond conjugates: the preparation and characterization
Šlegerová, Jitka ; Konvalinka, Jan (advisor) ; Vaněk, Ondřej (referee)
Nanodiamonds represent a member of vast group of carbon nanomaterials. They are characterized by chemical stability, biocompatibility, low toxicity and possibility of surface chemical modification that enables further conjugation of biomolecules, namely proteins. The most important property of nanodiamonds is easily detectable fluorescence resistant towards photobleaching. This luminescence is produced by so called (N-V) centers that can be formed by irradiation of nanodiamonds using high energy beams. Fluorescence of nanodiamonds is useful for construction of fluorescent labels and probes, cellular targeting and internalization, controlled drug delivery as well as enzyme immobilization. Two main connection modes are suitable for biomolecules' attachment: non-covalent and covalent. The first one has been utilized for decoration of nanodiamonds by various proteins, e.g. lysozyme, cytochrome c, neurotoxin or antigen. The second possibility has been demonstrated also on various proteins and, furthermore, on glycoproteins, oligonucleotides, vitamins or growth factor. During the immobilization of biomolecule on the nanoparticle surface it is crucial to keep its function, for example the catalytic activity in enzymes. For such purposes, the oxime ligation (connection of aldehyde presented on biomolecule...
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Structure and function of C-type lectin NK cell receptors studied by recombinant expression and protein crystallography
Vaněk, Ondřej
Department of Biochemistry, Faculty of Science, Charles University in Prague 2010 Structure and function of C-type lectin NK cell receptors studied by recombinant expression and protein crystallography Abstract of Ph.D. thesis Ondřej Vaněk Supervisor: Prof. RNDr. Karel Bezouška, DSc. Natural killer cells (NK cells) were found out for their ability to spontaneously kill certain allogeneic tumour cell lines, without any previous sensitization. NK cells are part of non- adaptive immune response with very short reaction time against pathogens such as viruses, intracellular bacteria, parasites, and they are responsible for elimination of certain tumour cells and thus they are able to fight against malignancy and formation of metastasis. Activity of NK cells is regulated by the balance between activation and inhibitory signals mediated by the NK cell surface receptors. From the structural point of view, the majority of NK cell surface receptors could be classified as the C-type lectin or immunoglobulin-like receptors. One of many C-type lectin subgroups are type II lymphocyte receptors that are expressed on the NK cell surface. This study had two main aims. The first one was to find suitable expression and purification systems for selected C-type lectin receptors of NK cells and the other one was to perform their...
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Structure and function of C-type lectin NK cell receptors studied by recombinant expression and protein crystallography
Vaněk, Ondřej ; Bezouška, Karel (advisor) ; Hrabal, Richard (referee) ; Bařinka, Cyril (referee)
Department of Biochemistry, Faculty of Science, Charles University in Prague 2010 Structure and function of C-type lectin NK cell receptors studied by recombinant expression and protein crystallography Abstract of Ph.D. thesis Ondřej Vaněk Supervisor: Prof. RNDr. Karel Bezouška, DSc. Natural killer cells (NK cells) were found out for their ability to spontaneously kill certain allogeneic tumour cell lines, without any previous sensitization. NK cells are part of non- adaptive immune response with very short reaction time against pathogens such as viruses, intracellular bacteria, parasites, and they are responsible for elimination of certain tumour cells and thus they are able to fight against malignancy and formation of metastasis. Activity of NK cells is regulated by the balance between activation and inhibitory signals mediated by the NK cell surface receptors. From the structural point of view, the majority of NK cell surface receptors could be classified as the C-type lectin or immunoglobulin-like receptors. One of many C-type lectin subgroups are type II lymphocyte receptors that are expressed on the NK cell surface. This study had two main aims. The first one was to find suitable expression and purification systems for selected C-type lectin receptors of NK cells and the other one was to perform their...
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Cloning, expression and biochemical characterisation of mouse glutamate carboxypeptidase II.
Knedlík, Tomáš ; Vaněk, Ondřej (referee) ; Konvalinka, Jan (advisor)
English Abstract Glutamate carboxypeptidase II (GCPII) is a membrane metallopeptidase expressed in many human tissues, predominantly in prostate, brain and small intestine. In brain it cleaves the most abundant peptide neurotransmitter N-acetyl-L-aspartyl-α-L-glutamate into N-acetyl-L-aspartate and free L-glutamate. Thus, GCPII participates in glutamate excitotoxicity through the release of free glutamate into the synaptic cleft. Inhibition of this activity has been shown to be neuroprotective in rats. In the human jejunal brush border, GCPII cleaves off terminal glutamate moieties from poly-γ-glutamylated folates, which can be then transported across the intestinal mucosa. The function of GCPII in human prostate is unknown but it is overexpressed in prostate cancer. Therefore, GCPII is an important marker of prostate cancer and its progression.Moreover, it could become a perspective target for treatment of prostate cancer as well as neuronal disorders associated with glutamate excitotoxicity. For the development and testing of novel drugs and therapeutics it is necessary to have an appropriate animal model. Mouse (Mus musculus) is such a model and it is widely used by many experimentators. However, no detailed comparison of mouse and human GCPII orthologs regarding their enzymatic activity, inhibition...
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Comparative study of body temperature measurement using various technologies
Vaněk, Ondřej ; Sekora, Jiří (referee) ; Provazník, Ivo (advisor)
The aim of this work is to compare the body temperature measurement with different contact thermometers. The theoretical part includes studying the topic, familiarization with the situation at the Faculty Hospital Brno, Bohunice and suggestion of method for comparison. The practical part includes thermometers measurement in the laboratory to verify the accuracy, clinical measurement on patients in different parts of the body and under various conditions and statistical evaluation of results.
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