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The development of new radical cross-linkers for structural characterization of proteins
Karpíšek, Michael ; Kukačka, Zdeněk (advisor) ; Chmelík, Josef (referee)
Proteins are important biomolecules because they carry out many essential functions. Since the structure of these biomolecules is usually closely linked to their function, there are numerous techniques to determine their three-dimensional structure. Exploiting the advantages of mass spectrometry, chemical cross-linking coupled to mass spectrometry is one of the methods that is used for structural characterization of studied molecule. Currently there is no reagent that targets and links all of the aromatic proteinogenic residues found in the sequence of proteins. In this work, we verified the reactivity of newly developed cross-linker towards aromatic residues and sulfhydryl group of cystein. This new reagent is based on the so-called Togni reagents that serve as trifluoralkylation agents. First, we monitored the impact of several different conditions on the composition of the reaction products using apomyoglobin as a model protein and electrosprey ionization as the method of choice. Subsequently we employed bottom up approach to find out which residues were modified. For this purpose we chose apomyoglobin and two other proteins that contain cysteine in its reduced form - RhoA and HSC70. Based on obtained results, we could confirm the ability of the new cross-linker to modify both aromatic residues...
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Traffic Violation Detection on Crossroads
Karpíšek, Miroslav ; Bartl, Vojtěch (referee) ; Špaňhel, Jakub (advisor)
This bachelor thesis presents procedure for the detection of red-light violation. In the theoretical part of the thesis, the current solution aproaches used in image processing are described. The practical part focuses on creation of program for automatic traffic corridors detection, vehicle tracking and the current traffic light state detection. The results obtained by experimenting with the proposed procedure and the possibilities of its further improvement are also discussed.
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System for Acquisition and Analysis of Real Estate Data from WWW
Karpíšek, Miroslav ; Hynek, Jiří (referee) ; Burget, Radek (advisor)
This project is focused on designing a system for automatic extraction of real estate data from real estate servers. In the theoretical part, this work is first placed in the context of data engineering. Subsequently, the used technologies and system components that were used in the project are described, same as their importance within the framework of the created platform intended for real estate investors for the purpose of renting them. The work also includes an evaluation of the basic methods used to predict the rent amount, which is used to calculate the return of investment.
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The development of new tyrosin binding cross-linkers for structural characterization of proteins
Karpíšek, Michael ; Kukačka, Zdeněk (advisor) ; Talacko, Pavel (referee)
Proteins are cornerstones of all living organisms. A lot of energy is invested in studying structures of proteins, because their function is determined by their structure. One of the techniques used to access the structure of proteins is chemical cross-linking in combination with mass spectrometry. In spite of their number, most of the cross-linkers bind few aminoacids such as lysine, cysteine or glutamic and aspartic acid. In our work we tried to develop tyrosin-binding cross-linkers. Using top down and bottom up approaches we studied the influence of N-hydroxysuccinimide esters and imidazole on the reactivity of the cross-linker in different pH on model proteins. According to the acquired data there is a difference in overal reactivity and specifity between the used cross-linkers. The cross-linkers containing imidazole modified tyrosine more although the difference was small and binding of tyrosine was not selective. [IN CZECH]
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Traffic Violation Detection on Crossroads
Karpíšek, Miroslav ; Bartl, Vojtěch (referee) ; Špaňhel, Jakub (advisor)
This bachelor thesis presents procedure for the detection of red-light violation. In the theoretical part of the thesis, the current solution aproaches used in image processing are described. The practical part focuses on creation of program for automatic traffic corridors detection, vehicle tracking and the current traffic light state detection. The results obtained by experimenting with the proposed procedure and the possibilities of its further improvement are also discussed.
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